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  • Lepidoptera  (1,054)
  • Immunocytochemistry  (856)
  • Springer  (1,910)
  • American Chemical Society (ACS)
  • 101
    ISSN: 1615-6102
    Keywords: Flax ; Cell wall ; Golgi apparatus ; β-Galactans ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Most cell wall components are carbohydrate including the major matrix polysaccharides, pectins and hemicelluloses, and the arabinogalactan-protein proteoglycans. Both types of molecules are assembled in the Golgi apparatus and transported in secretory vesicles to the cell surface. We have employed antibodies specific to β-(1→6) and β-(1→4)-D-galactans, present in plant cell wall polysaccharides, in conjunction with immunofluorescence and electron microscopy to determine the location of the galactan-containing components in the cell wall and Golgi stacks of flax root tip tissues. Immunofluorescence data show that β-(1→4)-D-galactan epitopes are restricted to peripheral cells of the root cap. These epitopes are not expressed in meristematic and columella cells. In contrast, β-(1→6)-D-galactan epitopes are found in all cell types of flax roots. Immunogold labeling experiments show that both epitopes are specifically located within the wall immediately adjacent to the plasma membrane. They are also detected in Golgi cisternae and secretory vesicles, which indicates the involvement of the Golgi apparatus in their synthesis and transport. These findings demonstrate that the synthesis and localization of β-(1→4)-D-galactan epitopes are highly regulated in developing flax roots and that different β-linked D-galactans associated with cell wall polysaccharides are expressed in a cell type-specific manner.
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  • 102
    ISSN: 1615-6102
    Keywords: Ferritin ; Nitrogen fixation ; Nodule development ; Plastid ; Legume ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In eukaryotic organisms ferritin is a protein involved in the storage of iron. The occurrence of ferritin and its relationship to the effectiveness of the nitrogen-fixing activity have been previously studied during the early stages of the nodule development by biochemical methods. We have used immunocytochemistry techniques to determine the precise location of ferritin and the behavior of this protein along the nodule development. The major localization was found in plastids and amyloplasts of infected and uninfected cells of the three legume nodules studied. A decrease of the immunolabelling was observed in infected cells of lupin and soybean senescing nodules and in the senescent zone of indeterminate alfalfa nodules. In the cortex of soybean and lupin nodules, ferritin increased during nodule ageing and the immunogold particles were mainly located in crystalline structures. The putative role of ferritin and plastids during nodule development is discussed.
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  • 103
    ISSN: 1432-0878
    Keywords: Key words Annexin 5 ; Immunocytochemistry ; Pituitary ; Ovary ; Testis ; Adrenal gland ; Thyroid gland ; Rat (wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Annexin 5, a unique calcium- and phospholipid-binding protein, has been investigated for its specific distribution in rat endocrine organs by immunocytochemistry with a specific antiserum to recombinant rat annexin 5. Follicular epithelial cells and parafollicular cells of the thyroid gland, adrenocortical cells of the zona fasciculata and zona reticularis, luteal cells, testicular interstitial cells, and Sertoli cells were shown to contain annexin 5. To examine whether the synthesis of annexin 5 would be affected by a change in humoral signal, the distribution of annexin 5 in the anterior pituitary was examined three weeks after ovariectomy. The withdrawal of ovarian hormones induced huge castration cells in the anterior pituitary gland, which contained abundant annexin 5. Annexin 5 was not detected in the pineal gland, the parathyroid gland, the islet of Langerhans, the adrenal medulla, zona glomerulosa cells, and granulosa cells. Since annexin 5 was shown to exist in many of the endocrine tissues examined, to be localized in specific cell types, and to be abundant in castration cells, it is suggested that annexin 5 contributes to secretory cell functions, which may be common to endocrine cells secreting chemically different hormones.
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  • 104
    ISSN: 1432-0878
    Keywords: Key words Fish ; Zebrafish ; Immunocytochemistry ; Keratin ; Cytoskeleton ; Danio rerio (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have identified a number of type I and type II keratins in the zebrafish Danio rerio by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot-binding assay and immunoblotting. These keratins range from 56 kDa to 46 kDa in molecular mass and from pH 6.6 to pH 5.2 in isoelectric point. Type II zebrafish keratins exhibit significantly higher molecular masses (56–52 kDa) compared with the type I keratins (50–48 kDa), but the isoelectric points show no significant difference between the two keratin subclasses (type II: pH 6.0–5.5; type I: pH 6.1–5.2). According to their occurrence in various zebrafish tissues, the identified keratins can be classified into “E” (epidermal) and “S” (simple epithelial) proteins. A panel of monoclonal anti-keratin antibodies has been used for immunoblotting of zebrafish cytoskeletal preparations and immunofluorescence microscopy of frozen tissue sections. These antibodies have revealed differential cytoplasmic expression of keratins; this not only includes epithelia, but also a variety of mesenchymally derived cells and tissues. Thus, previously detected fundamental differences in keratin expression patterns between higher vertebrates and a salmonid, the rainbow trout Oncorhynchus mykiss, also apply between vertebrates and the zebrafish, a cyprinid. However, in spite of notable similarities, trout and zebrafish keratins differ from each other in many details. The present data provide a firm basis from which the application of keratins as cell differentiation markers in the well-established genetic model organism, the zebrafish, can be developed.
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  • 105
    ISSN: 1432-0878
    Keywords: Key words Histamine ; Immunocytochemistry ; Mast cells ; Melanocytes ; Nerves ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The number, distribution, and ultrastructural characteristics of mast cells were assessed in the tongue, heart, and kidney of the frog Rana esculenta. The density of tongue mast cells (253±45 mast cells/mm2) was significantly higher than that of the heart (5.3±0.4/mm2) and kidney (15.3±1.4 /mm2). A striking feature of this study was the remarkable association of frog mast cells to nerves. The ultrastructural study of the mast cell/nerve association demonstrated that mast cells were closely apposed to or even embedded in nerves. Mast cells were also physically associated with melanocytes even in the heart. Mast cells were Alcian blue+/safranin+ in the tongue and in the peritoneum, whereas in the heart and in the kidney they were Alcian blue–/safranin+. The mast cells in the lamina propria of the gastrointestinal tract were Alcian blue+/safranin–. The cytoplasm of frog mast cells was packed with numerous heterogeneous, membrane-bound granules. The ultrastructure of these cytoplasmic granules was unique, being totally unlike any other previously described granules in other animal species as well as in man. The histamine content/frog mast cell (≈0.1 pg/cell) was approximately 30 times lower than that of human mast cells isolated from different tissues (≈3 pg/cell). A monoclonal anti-histamine antibody was used to confirm the ultrastructural localization of histamine in secretory granules in frog mast cells.
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  • 106
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    Biodiversity and conservation 7 (1998), S. 725-748 
    ISSN: 1572-9710
    Keywords: farm woodlands ; island biogeography ; Lepidoptera ; moths ; species richness ; woodlands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Farm Woodland Scheme, which provided incentives to convert agricultural land to timber production, contained an implicit assumption that farm woodlands produce important benefits for wildlife. The moth fauna of 18 farm woodlands in the Vale of York was surveyed between May and November 1991. The aims were twofold. The first was to determine if there were benefits for moth species diversity. The second was to ascertain whether concepts of island biogeography and the plant species richness of the woods were related to the moth species composition. Eleven families, 214 species and over 16 000 individuals of moths were recorded. Classification of the species presence/absence matrix indicated that small woods (less than 1ha) did not have characteristic woodland moth communities. Woods larger than 5ha were judged to be more valuable for the long-term conservation of woodland moth diversity. The best predictor of moth species richness was the herbaceous plant species richness within woodlands. Species richness of the family Geometridae was positively related to woodland area, as well as to woodland shape (compact shapes being preferable to elongated shapes). Characteristic woodland species are influenced by isolation (less isolated woods being richer in species). The implications of different powers of dispersal between moth families are discussed. Farm woodlands will be of more value for the conservation of the Macrolepidoptera if they are large, compact and incorporate remnants of existing woodland with extant herbaceous vegetation. These should be factors which are taken into consideration when providing incentives to establish and manage farm woodlands.
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  • 107
    ISSN: 1572-9710
    Keywords: Lepidoptera ; biogeography ; islands ; migration ; colonization ; metapopulations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Records of Hipparchia semele on British and Irish islands have been modelled against island area, isolation (sea and land distance) and the size of the nearest potential source populations. All three variables have been found to contribute significantly to the presence or absence of H. semele on the islands. Isolation is a more significant predictor than island area. This result differs from the multiple species case where area was found to be a more important influence than isolation. Records on islands are also shown to depend on the size of populations at the nearest sources; this underpins the relationships identified for the multiple species case, first, between the number of species on islands and at nearest sources and, second, between the incidence of species on islands and at nearest sources. There are clear indications that smaller islands may become increasingly marginalized for H.␣semele; with ongoing habitat loss, because isolation increases and source populations become sparser, the probability of H. semele recolonizing islands also decreases.
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  • 108
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    Biodiversity and conservation 7 (1998), S. 967-979 
    ISSN: 1572-9710
    Keywords: altitude ; biodiversity ; demographic stability ; Lepidoptera ; northern Spain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Quick studies on biodiversity are frequently used in conservation assessments. Conclusions from these studies would be consistent if assemblages under consideration are stable over time. The stability of butterfly assemblages in the Picos de Europa in northern Spain was examined at several levels of numerical resolution. The survey was carried out in 1993 and 1995, which allowed at least one complete turnover of all individuals in each locality. Butterfly assemblages were usually stable at the levels of number of species and total number of individuals, species presence and absence, and abundance rankings. But, absolute abundances of individual species changed from 1993 to 1995 out of synchrony with one another. Regional distributions and altitudinal ranges of species were also stable. Assemblages were similar in both study years, judging by similar site ordinations by reciprocal averaging. Overall, these results are in accordance with most studies where stability has been examined at several numerical levels; they also suggest that abundances of individual species do not vary enough to disrupt overall assemblage abundance rankings. Butterfly assemblages at sites at higher altitude tended to be more unstable in terms of abundance rankings and absolute abundances. This agrees with ecological theory predicting less stable assemblages in physically ‘harsh’ environments. We concluded that monitoring for a relatively short time period can give a clear picture of both local and regional butterfly biodiversity and species composition.
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  • 109
    ISSN: 1572-9710
    Keywords: community indices ; Lepidoptera ; randomization ; species richness ; transect counts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Community indices were evaluated at different frequency of transect counts of butterflies by randomization. Randomized samples were obtained by re-arranging 1, 3, 6, 12 and 18 counts from 18 counts at a study site (Hiraoka), and 1, 2, 3, 5 and 15 from 15 at another site (Narukawa), in 1994. Morisita's new estimate of species richness, QM gave a good estimate when the number of counts was five or more. QM estimated from the sum of both assemblages (58 species) was 70.7, while 73 species were recorded at least once from 1990 to 1994. The Octave method always resulted in an underestimation. Diversity indices, Shannon's H′, Simpson's D and β, Hurlbert's E(Sn), Itow's b, and Morisita's H⊛′ gave good estimates when the number of counts was five or more. Temporariness of univoltine species in flight period made a bias in the estimates. We concluded that the count is needed at least six times (once a month) and the estimates were improved by separating the multivoltine species from the univoltine species. Intensive counting during the flight period of the univoltine species was recommended.
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  • 110
    ISSN: 1615-6102
    Keywords: Lupinus albus ; Nitrogen fixation ; Oxygen diffusion ; Glycoprotein ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The monoclonal antibodies MAC236 and MAC265, raised against a soluble component of pea nodules, were used to elucidate the presence and subcellular localization of glycoprotein epitopes during the development of lupin (Lupinus albus L. cv. Multolupa) nodules, by means of immunocytochemistry and Western blot analysis. These antibodies recognize a single band of 95 kDa in pea, soybean and bean nodules, whilst two different bands of 240 and 135 kDa cross-react with MAC236 and MAC265 respectively in lupin nodules. This fact may indicate that the recognized epitopes can be present in different subcellular compartments and/or play different roles through the development of functional nodules. The results show that MAC265 is mainly associated with Bradyrhizobium infection and with the development of nodule primordium, in the first stages of nodulation. MAC265 is also detected when glycoprotein transport takes place across the cytoplasm and the cell wall, and also in the intercellular spaces of the middle cortex, attached to cell walls. The amount of MAC265 remains constant through nodule development. In contrast the amount of MAC236 increases with nodule age, parallel to the establishment of nitrogenase activity. This antibody is localized in cytoplasmic globules attached to the inner side of cell walls in the middle cortex, and mainly in the matrix filling the intercellular spaces of the middle and inner cortex. This main site of localization of MAC236 may indicate a role in the functioning of the oxygen diffusion barrier.
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  • 111
    ISSN: 1615-6102
    Keywords: Chloroplast development ; Cotton ; Fluorescence induction kinetics ; Ultrastructure ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Many of the studies of chloroplast ontogeny in higher plants have utilized suboptimal conditions of light and growth to assess development. In this study, we utilized structural, immunological, and physiological techniques to examine the development of the chloroplast in fieldgrown cotton (Gossypium hirsutum cv. “MD 51 ne”). Our youngest leaf sample developmentally was completely folded upon itself and about 0.5 cm in length; leaves of this same plastochron were followed for three weeks to the fully expanded leaf. The chloroplasts at the earliest stage monitored had almost all of the lamellae in small, relatively electron-opaque grana, with relatively few thylakoids which were not appressed on at least one surface. During the development of the thylakoids, the membranes increase in complexity, with considerable stroma lamellae development and an increase in the number of thylakoids per granum. Besides the increase in complexity, both the size and numbers of the chloroplast increase during the development of the leaf. Developmental changes in six thylakoid proteins, five stromal proteins, and one peroxisomal protein were monitored by quantitative immunocytochemistry. Even at the earliest stages of development, the plastids are equipped with the proteins required to carry out both light and dark reactions of photosynthesis. Several of the proteins follow three phases of accumulation: a relatively high density at early stages, a linear increase to keep step with chloroplast growth, and a final accumulation in the mature chloroplast. Photosystem-II(PS II)-related proteins are present at their highest densities early in development, with an accumulation of other parts of the photosynthetic apparatus at a latter stage. The early accumulation of PS-II-related proteins correlates with the much lower ratio of chlorophylla tob in the younger leaves and with the changes in fluorescence transients. These data indicate that some of the conclusions on chloroplast development based upon studies of intercalary meristems of monocots or the greening of etiolated plants may not be adequate to explain development of chloroplasts in leaves from apical meristems grown under natural conditions.
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  • 112
    ISSN: 1432-0878
    Keywords: Key words: Neurofilaments ; Phosphorylation ; Differentiation ; Immunocytochemistry ; Brain storage ; Fixation ; Microwave ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurofilaments, which are exclusively found in nerve cells, are one of the earliest recognizable features of the maturing nervous system. The differential distribution of neurofilament proteins in varying degrees of phosphorylation within a neuron provides the possibility of selectively demonstrating either somata and dendrites or axons. Non-phosphorylated neurofilaments typical of somata and dendrites can be visualized with the aid of monoclonal antibody SMI 311, whereas antibody SMI 312 is directed against highly phosphorylated axonal epitopes of neurofilaments. The maturation of neuronal types, the development of area-specific axonal networks, and the gradients of maturation can thus be demonstrated. Optimal immunostaining with SMI 311 and SMI 312 is achieved when specimens are fixed in a mixture of paraformaldehyde and picric acid for up to 3 days and sections are incubated free-floating. Neurons, with their dendritic domains immunostained by SMI 311 in a Golgi-like manner, can be completely visualized in relatively thick sections. The limitations of Golgi-preparations, such as glia-labeling, artifacts, and the staining of only a small non-representative percentage of existing neurons, are not apparent in SMI preparations, which additionally provide the possibility of selectively staining axonal networks. The results achieved in normal fetal brain provide the basis for studies of developmental disturbances.
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  • 113
    ISSN: 1432-0878
    Keywords: Key words Co-expression of mRNAs ; Gonadotropin-releasing hormone ; Immunocytochemistry ; In situ hybridization ; Oncorhynchus nerka (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The localization of two salmon-type gonadotropin-releasing hormone (sGnRH) precursors, pro-sGnRH-I (short type) and pro-sGnRH-II (long type), was investigated by using in situ hybridization techniques in the brain of the landlocked sockeye salmon, Oncorhynchus nerka. We used 30-mer oligonucleotide probes complementary to pro-sGnRH-I and pro-sGnRH-II cDNA. No significant differences were observed in the localization of sGnRH neurons expressing pro-sGnRH-I and pro-sGnRH-II mRNAs; both were expressed in the olfactory nerve, the olfactory bulbs, the regions between the olfactory bulb and telencephalon, the ventral telencephalon, the preoptic area, and the hypothalamus. Almost all sGnRH neurons examined co-expressed both precursors. The expression of two sGnRH precursors in the same neuron and the wide distribution of such neurons in the brain suggest that there are no functional differences between the two precursors.
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  • 114
    ISSN: 1432-0878
    Keywords: Key words Salmon GnRH ; Chicken GnRH ; II ; Radioimmunoassay ; Immunocytochemistry ; In situ hybridization ; Oncorhynchus masou (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Ontogenic development of salmon gonadotropin-releasing hormone (GnRH) and chicken GnRH-II systems in masu salmon (Oncorhynchus masou) was examined. Salmon GnRH was first detected by radioimmunoassay in the embryo on day 36 after fertilization. Salmon GnRH-immunoreactive fibers were detected initially by immunocytochemistry in the vicinity of the olfactory placode of the embryo (day 36) and were distributed widely in the brain as well as in the pituitary gland of fish just after hatching (day 80). Salmon GnRH-immunoreactive neuronal somata were first detected about 6 months after fertilization in the rostroventral brain area, ranging from the olfactory nerve to the preoptic area. Salmon GnRH neuronal somata were detected earlier by in situ hybridization than by immunocytochemistry. Neuronal somata expressing salmon GnRH mRNA were first detected in the vicinity of the olfactory epithelium on day 40 and then were seen to be migrating from the olfactory epithelium, along the olfactory nerve, on day 60 and in the transitional area between olfactory nerve and olfactory bulb on day 80. In the brain, these neurons were first detected in the ventral olfactory bulb on day 80, and thereafter they were also detected in the caudal brain regions. The chicken GnRH-II system was detected later than the salmon GnRH system; chicken GnRH-II was first detected by radioimmunoassay on day 57, and chicken GnRH-II-immunoreactive fibers were first detected on day 67. Chicken GnRH-II-immunoreactive neuronal somata were not detected during the observation period. These results suggest that salmon GnRH neurons derive from the olfactory placode and then migrate into the brain and that salmon GnRH is synthesized before chicken GnRH-II.
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  • 115
    ISSN: 1432-0878
    Keywords: Key words Adrenomedullin ; Pancreas ; Development ; Immunocytochemistry ; Colocalizations ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Adrenomedullin is an α-amidated 52-amino acid peptide involved in many physiological actions, among others the regulation of insulin secretion. Using immunohistochemical methods, we found that adrenomedullin immunoreactivity first appears at day 11.5 of embryonic development in the rat, coinciding with the appearance of pancreatic glucagon. The early appearance of adrenomedullin in the developing pancreas may indicate an active involvement in either the morphogenesis of the organ or its endocrine/paracrine/autocrine hormone regulation during intrauterine life. We also investigated the pattern of colocalizations of adrenomedullin with the other pancreatic hormones. At some point during development all the cell types express adrenomedullin, progressively evolving towards the adult pattern where only the pancreatic polypeptide cells contain a strong immunoreactivity for adrenomedullin. At this point the remaining cells of the islet are, in general, weakly stained. This sequential and time-dependent expression of adrenomedullin suggests a tight regulation similar to that observed for other modulatory substances responsible for embryonic morphogenesis.
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  • 116
    ISSN: 1432-0878
    Keywords: Key words Pepsinogen C ; Ontogeny ; Mucous neck cell ; Chief cell ; Intermediate mucopeptic cell ; Immunocytochemistry ; In situ hybridization ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The ontogeny of pepsinogen C-producing cells in rat fundic glands was studied by means of light and electron microscopy using an antiserum raised against a synthetic peptide based on rat pepsinogen C. To confirm the immunocytochemistry results, the expression of rat pepsinogen C messenger RNA (mRNA) in the fundic gland was also examined by in situ hybridization using a digoxigenin-labeled RNA probe. In adult rats, pepsinogen C was produced by chief cells, mucous neck cells, and intermediate mucopeptic cells. Pepsinogen C-producing cells appeared in embryos as early as 18.5 days’ gestation. The development of these cells could be classified into four stages: (1) 18.5 days’ gestation to 0.5 days after birth; (2) 0.5 days to 2 weeks after birth; (3) 3–4 weeks after birth; (4) 4–8 weeks after birth. In embryos and young animals, pepsinogen C-producing cells were mucopeptic cells. By 4 weeks after birth, mucous neck cells could be distinguished morphologically. The maturation stages of the chief cells could be traced by electron microscopy along the longitudinal axis of the rat fundic gland by double-staining with anti-pepsinogen C antibody and periodic acid-thiocarbohydrazide-silver proteinate. Positive reactions for pepsinogen C and pepsinogen C mRNA expression were detected in mucous neck cells. Therefore, we conclude that mucous neck cells are precursor cells of chief cells. Mucous neck cells, intermediate cells, and chief cells are in the same differentiating cell lineage.
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  • 117
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    Cell & tissue research 293 (1998), S. 245-252 
    ISSN: 1432-0878
    Keywords: Key words Synaptic-like microvesicles ; Synaptophysin ; Synaptobrevin ; Immunocytochemistry ; Immunoelectron microscopy ; Meriones unguiculatus (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Mammalian pinealocytes contain several synaptic membrane proteins which probably play a role in the targeting and exocytosis of secretory vesicles, in particular of synaptic-like microvesicles (SLMVs). The latter are considered as the endocrine equivalent of neuronal synaptic vesicles. By means of immunocytochemical techniques and immunoblot analyses, we now show that two further key components of the molecular apparatus regulating neurotransmitter release are present in the gerbil pineal gland, i.e., munc-18–1 and cysteine string protein (csp). In addition to varicosities of nerve fibres, munc-18–1 and csp could be localized to pinealocytes where both proteins were markedly enriched in process swellings. When using antibodies against csp for an immunogold electron-microscopic study of pinealocytes, gold particles consistently decorated profiles of pleomorphic SLMVs. Interestingly, we found that also the cytosolic protein munc-18, which is partially recruited to the plasmalemma in neurons, was associated to a significant extent with SLMVs of pinealocytes and synaptic vesicles of neurons, respectively. This localization implies that munc-18 at least partially exerts its regulatory functions while being bound to secretory vesicle membranes. Our results indicate that in endocrine cells such as pinealocytes the synaptic proteins munc-18–1 and csp play essential roles during the life cycle of SLMVs.
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  • 118
    ISSN: 1432-0878
    Keywords: Key words Enamelin ; Immunocytochemistry ; Amelogenesis ; Tooth development ; Enamel ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Enamelins comprise an important family of the enamel matrix proteins. Porcine tooth germs were investigated immunochemically and immunocytochemically using two antibodies: a polyclonal antibody raised against the porcine 89-kDa enamelin (89 E) and an affinity purified anti-peptide antibody against the porcine enamelin amino-terminus (EN). Immunochemical analysis of layers of immature enamel from the matrix formation stage detected immunopositive protein bands ranging from 10 kDa to 155 kDa in the outer layer enamel sample irrespective of the antibodies used. In contrast, the middle and inner enamel layer mainly contained lower molecular weight enamelins. In immunocytochemical analyses of the differentiation stage, 89 E stained enamel matrix islands around mineralized collagen fibrils of dentin, while EN stained both enamel matrix islands and stippled material. At the matrix formation stage, both antibodies intensely stained enamel prisms located in the outer layer. In the inner layer, 89 E moderately stained enamel matrix homogeneously, while EN primarily stained the prism sheath. The intense immunoreaction over the surface layer of enamel matrix at the matrix formation stage, following staining with 89 E and EN, disappeared by the end of the transition stage and the early maturation stage, respectively. The Golgi apparatus and secretory granules in the ameloblasts from the late differentiation stage to the transition stage were immunostained by both antibodies. These results suggest that expression of enamelin continues from late differentiation to the transition stage and the cleavage of N-terminal region of enamelin occurs soon after secretion. Some enamelin degradation products, which apparently have no affinity for hydroxyapatite crystals, concentrate in the prism sheaths during enamel maturation.
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  • 119
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    Cell & tissue research 293 (1998), S. 349-355 
    ISSN: 1432-0878
    Keywords: Key words Germ cells ; Nucleus ; Ribonucleoproteins ; Immunocytochemistry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The convoluted cords present in the nuclei of rabbit primary spermatocytes and spermatids differ from previously described nuclear bodies. They are composed of proteic strands decorated with granules and, in most cases, are embedded in clusters of interchromatin granules. They are partly sensitive to trypsin and can be visualised with protein-specific stains. The decorating granules are similar in size and aspect to interchromatin granules. However, only the latter are continuously immunolabelled with anti-snRNPs (small ribonucleoproteins) antibodies during spermatogenesis. The complexity and organisation of the convoluted cords are modified specifically during cell differentiation. They might be involved in the storage, transport and release of interchromatin granules in male germ cells.
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  • 120
    ISSN: 1432-0878
    Keywords: Key words Plant lectins ; Epithelial cells ; Immunocytochemistry ; Autoradiography ; Immunoelectron microscopy ; Potato leafhopper ; Empoasca fabae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To identify the means by which phytohemagglutinin (PHA) exerts its toxicity on the potato leafhopper, four different methods (thick and semi-thin sectioning combined with immunofluorescent staining, in vitro receptor autoradiography, and immunoelectron microscopy) were used to elucidate the PHA target tissue, binding site, and its effects on this tissue. Sixteen 1- or 2-day-old female potato leafhoppers were fed for 36 h on each of three treatments: a control, diet or a diet containing either the PHA-E subunit or the PHA-L subunit. The PHA-E subunit, but not PHA-L, had previously been shown to be lethal. The insects were then prepared for both light and confocal microscopy. Analysis of images showed that PHA bound only to the surface of midgut epithelial cells of the potato leafhopper. PHA-E caused severe disruption, disorganization, and elongation of the brush border microvilli, and swelling of the epithelial cells into the lumen of the gut, leading to complete closure of the lumen. Furthermore, PHA-E stimulated the division of midgut epithelial cell nuclei, leading to two nuclei in each cell. Nuclei later elongated and degraded. In contrast, PHA-L had little effect on the epithelial cells of the midgut. It did not strongly bind to the surface of epithelial cells and caused much less disruption of brush-border microvilli, less disorganization of the cells and less elongation of nuclei. Strong binding of PHA occurred solely on the cell membrane of the brush border microvilli of epithelial cells. In contrast, the controls (i.e., midgut tissue, blocking agent, PHA, and antibodies) showed that midgut tissue was not autofluorescent and showed no fluorescent binding signal. Analysis of both bright- and dark-field images obtained by autoradiography and immunoelectron microscopy confirmed these findings.
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  • 121
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    Plant systematics and evolution 212 (1998), S. 159-176 
    ISSN: 1615-6110
    Keywords: Lamiaceae ; Cedronella canariensis ; Apidae ; Bombus canariensis ; Lepidoptera ; Macroglossa stellatarum ; Canary Islands ; cross-pollination ; gynodioecy ; gynomonoecy male sterility ; mixed mating system ; nectar-robbing ; self-pollination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract On the Canary Islands, we studied reproductive ecology of the perennial laurel forest herbCedronella canariensis (Lamiaceae). Flowers contained small quantities of concentrated nectar. Flower visitors were mainly bumblebees (Bombus terrestris subsp.canariensis) and lepidopterans (especiallyMacroglossa stellatarum). Their abundance, pollen load, and behaviour are reported.Cedronella canariensis was facultatively autogamous. The P:O ratio was higher than expected for a plant with a breeding system of this nature. Seed set was not pollen-limited. Selfed and outcrossed seeds differed in weight. Small, lightly coloured flowers with dysfunctional stamens appeared towards the end of the season. This male-sterility might be induced by drought.
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  • 122
    ISSN: 1432-0878
    Keywords: Key words Pancreas ; exocrine ; Hibernation ; Amylase ; Immunocytochemistry ; Muscardinus avellanarius (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Pancreatic acinar cells of euthermic, hibernating and arousing individuals of the hazel dormouse Muscardinus avellanarius (Gliridae) have been observed at the electron-microscopic level and analysed by means of ultrastructural morphometry and immunocytochemistry in order to investigate possible fine structural changes of cellular components during periods of strikingly different degrees of metabolic activity. During hibernation, the cisternae of the rough endoplasmic reticulum (RER) flatten assuming a parallel pattern, the Golgi apparatus is extremely reduced and the mitochondria contain many electron-dense particles. The cell nuclei appear irregularly shaped, with deep indentations containing small zymogen granules. They also contain abundant coiled bodies and unusual constituents, such as amorphous bodies and dense granular bodies. Large numbers of zymogen granules occur in all animals. However, the acinar lumina are open and filled with zymogen only in euthermic animals, whereas, in hibernating and arousing individuals, they appear to be closed. Morphometrical analyses indicate that, in pancreatic acinar cells, nuclei and zymogen granules significantly decrease in size from euthermia to hibernation, probably reflecting a drastic decrease of metabolic activities, mainly protein synthesis and processing. In all the studied animals, immunocytochemistry with specific antibodies has revealed an increasing gradient in α-amylase content along the RER-Golgi-zymogen granule pathway, reflecting the protein concentration along the secretory pathway. Moreover, during deep hibernation, significantly larger amounts of α-amylase accumulate in RER and zymogen granules in comparison to the other seasonal phases analysed. Upon arousal, all cytoplasmic and nuclear constituents restore their euthermic aspect and all morphometrical and immunocytochemical parameters exhibit the euthermic values, thereby indicating a rapid resumption of metabolic activities.
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  • 123
    ISSN: 1432-0878
    Keywords: Key words Electron microscopy ; F-actin ; Freeze-fracture ; Immunocytochemistry ; Water permeability ; Antidiuretic hormone ; Urinary bladder ; Rana temporaria (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Electron and confocal microscopy, using immunocytochemical methods, was employed to assess osmotic water permeability of the frog (Rana temporaria) urinary bladder during transcellular water transport, induced by antidiuretic hormone (ADH) or by wash-out of autacoids from serosal, ADH-free Ringer solution. The increase of osmotic water permeability of the urinary bladder was accompanied by relevant ultrastructural changes, the most remarkable being: (1) the appearance of aggregates of intramembranous particles in the apical membrane of granular cells, and the extent of the membrane area covered by the aggregates proportional to that of the water flow; (2) redistribution of actin filaments in the cytoplasm of granular cells; judging from the anti-actin label density, the number of actin filaments in the apical region of cytoplasm was reduced by 2.5–4 times compared with normal; (3) a decrease in the total electron density of the cytoplasm due to the increased water content of granular cells.
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  • 124
    ISSN: 1432-0878
    Keywords: Key words Enteric nervous system ; Somatostatin ; Immunocytochemistry ; Intestinal motility ; Synaptic connections ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Somatostatin immunoreactivity occurs in a specific subgroup of cholinergic descending interneurons in the myenteric plexus of the guinea-pig small intestine. In the present work, we made light- and electron-microscopic investigations of chemically defined inputs to these neurons, in order that the origins of the connections of other neurons with them could be deduced. Somatostatin-immunoreactive synapses and close contacts were found on the cell bodies and filamentous processes of somatostatin neurons; these were 84% of all inputs. It is thus confirmed that this class of interneuron forms chains that project anally. Descending interneurons with immunoreactivity for nitric oxide synthase provided 14% of inputs to somatostatin-immunoreactive descending interneurons. An antiserum against a calcium-binding protein, calbindin, was used as marker for the majority of intrinsic primary afferent neurons, AH/Dogiel type II neurons; this class of neurons provided only 2.5% of the inputs to somatostatin-immunoreactive descending interneurons. We conclude that somatostatin-immunoreactive descending interneurons are involved in the conduction of impulses distally along the full length of the small intestine, but receive only a minor input from calbindin-immunoreactive primary afferent neurons.
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  • 125
    ISSN: 1432-0878
    Keywords: Key words Insect nervous system ; Immunocytochemistry ; Neural development ; Neuropeptide ; Neurohormone ; Locustatachykinin ; Spodoptera litura (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neuropeptides with similarities to vertebrate tachykinins, designated tachykinin-related peptides (TRPs), have been identified in several insect species. In this investigation we have utilized an antiserum raised to one of the locust TRPs, locustatachykinin-I (LomTK-I), to determine the distribution pattern of LomTK-like immunoreactive (LTKLI) neurons in the developing nervous system of the moth Spodoptera litura. A number of LTKLI neurons could be followed from the larval to the adult nervous system: a set of median neurosecretory cells (MNCs) in the brain, a pair of brain descending neurons and a few sets on neurons in the ventral nerve cord. The distribution of LTKLI neurons in the adult brain is very similar to that seen in other insect species with prominent arborizations in the central body, antennal lobes, mushroom body calyces, optic lobe neuropils and other distinct neuropil areas in the protocerebrum and tritocerebrum. A new finding is the presence of LTKLI neurosecretory cells with axon terminals in the anterior aorta and corpora cardiaca, suggesting for the first time a neurohormonal role of tachykinin-related peptide(s) in insects. During postembryonic development the number of LTKLI neurons in the ventral nerve cord decreases somewhat, whereas the number increases in the brain. Thus the functional roles of TRPs may change to some extent during development.
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  • 126
    ISSN: 1432-0878
    Keywords: Key words Pancreas ; Stomach ; Duodenum ; Ribonuclease protection assay ; Immunocytochemistry ; Protease ; Rat ; (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Prior studies have revealed the presence of chymotrypsinlike protease in peripheral organs, although no definitive evidence for the synthesis of this enzyme in tissue other than the pancreas is available. In an attempt to detect chymotrypsinogen mRNA in peripheral organs, a fragment of the pancreatic chymotrypsin mRNA from rat was amplified using PCR. The sequence was identified as a portion of the rat chymotrypsin B gene overlapping exon 5 through exon 7. It was subcloned into the pGEM-4Z vector and used as a template for the vitro transcription of an antisense riboprobe. Using ribonuclease protection and Northern blot analyses, chymotrypsin mRNA was detected in the rat pancreas, stomach, duodenum, ovary, and spleen. Monoclonal and polyclonal antisera against chymotrypsin detected chymotrypsinlike immunoreactivity in rat and human pancreas, rat stomach, duodenum and jejunum. Electrophoresis and immunoblotting revealed chymotrypsin-chymotrypsinogen bands (25–29 kDa) in the stomach and duodenum. Synthesis of a potent protease such as chymotrypsin in tissue other than pancreas is significant, suggesting a potential physiological and/or pathological role in these tissues.
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  • 127
    ISSN: 1432-0878
    Keywords: Key words Acrosome ; Epididymal maturation ; Monoclonal antibody ; Immunocytochemistry ; Spermatozoa ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have previously shown that a 90-kDa intra-acrosomal antigen, MN7, is restricted to the anterior acrosomal region of mouse, rat, and hamster spermatozoa. The present study has examined the localization and the behavior of MN7 during sperm maturation in the epididymis of the guinea pig by immunoelectron microscopy. MN7 showed not only a specific localization in the apical segment of the guinea pig sperm acrosome, but also a distinct alteration during maturation, as follows. MN7 was exclusively found both at the dorsal matrix and on the outer acrosome membrane (OAM)/matrix-associated materials in the apical segment. MN7 was initially distributed throughout the electron-lucent dorsal matrix in immature sperm but, during maturation, became more restricted to the spherical bodies within the electron-lucent area. MN7 on OAM/matrix-associated materials was first distributed along the ventral margin and the small area posterior to the dorsal matrix but, during maturation, disappeared from the ventral margin and became restricted to the dorsal region. These results indicate that MN7 is a good tool for studying the stepwise maturation of epididymal spermatozoa.
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  • 128
    ISSN: 1432-0878
    Keywords: Key words Mammalian-type thyrotropin ; Pituitary ; Immunocytochemistry ; Australian lungfish ; Neoceratodus forsteri (Dipnoi)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The binding sites of polyclonal antisera raised against the β-subunit of human thyroid-stimulating hormone (hTSHβ), hTSH, and ovine TSH (oTSH) have been localized in the pituitary gland of the Australian lungfish, Neoceratodus forsteri, using light microscopy. Reactivity toward anti-TSH antiserum was demonstrated in a slightly elongated and irregularly-shaped distinct cell type forming clusters in the dorso-central and ventral regions of the distal lobe. Their granules react with alcian blue (AB), and with periodic acid–Schiff (PAS), and after AB-PAS-orange G they stain blue or purple. The specificity of the different antisera was established by liquid-phase absorptions and confirmed in positive and negative tissue control systems. Our observations confirm that dipnoan (Neoceratodus) TSH shares a number of antigenic determinants with those of mammalian TSHβ and support the concept that mammalian TSHβ, or part of it, was established early in evolution, and that dipnoans (Neoceratodus) as living sarcopterygians may have an ancestor in common with the early amphibians. The mapping and detailed description of TSH-like immunoreactive cells may furnish a background to facilitate current and future analysis of the ontogeny and time course of TSH production and release in Neoceratodus in relation to different physiological conditions.
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  • 129
    ISSN: 1432-1939
    Keywords: Key words Insect-plant interactions ; Lepidoptera ; Polyphagy ; Specialization ; Host plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We analyzed geographic differentiation in oviposition preference in the anise swallowtail butterfly, Papilio zelicaon Lucas, which is one of the most widely distributed and polyphagous butterflies in western North America. Among 13 populations that span 1200 km of the range of P. zelicaon in the Pacific Northwest of North America, the overall oviposition preference hierarchy has not diverged significantly, even though these populations differ in the plant species they use in the field. The results indicate that differences in host availability and use have not favored major reorganizations in the preference hierarchy of ovipositing females. Instead, this butterfly has a conserved preference hierarchy that varies within a narrow range among populations. All populations ranked the four test plant species in the same overall relative order, even though these populations differ in the plant species they use in the field.
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  • 130
    ISSN: 1570-7458
    Keywords: in vitro rearing ; Campoletis sonorensis ; Hymenoptera ; Ichneumonidae ; Heliothis virescens ; Lepidoptera ; Noctuidae ; artificial medium ; sugars ; amino acids ; proteins ; luminosity ; medium volume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Campoletis sonorensis is a solitary larval endoparasitoid of several Heliothis spp. pests. This study was carried out to develop media devoid of insect sources for in vitro rearing the parasitoid. Trehalose, lysine, threonine, asparagine, glutamine, hydroxyproline, serine, bovine serum albumin, and lactalbumin were beneficial to C. sonorensis. Addition of fresh chicken egg yolk at a low level improved the artificial media. Addition of 20-hydroxyecdysone increased the molting rate, reduced the critical size at molt and decreased development time. Parasitoid development in vitro was dependent on eggs that had been in the host for at least 22 h. Luminosity was also critical for the development of C. sonorensis in vitro. Optimal development occurred in a L14:D10 photoperiod at 43 Lux light intensity. Utilizing the best media and conditions, 100% of the parasitoid larvae reached the second instar and over 37% molted to the third instar. However, no further development occurred.
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  • 131
    ISSN: 1570-7458
    Keywords: Sesamia nonagrioides ; Lepidoptera ; Noctuidae ; sex pheromone ; pest monitoring ; electrophysiological behaviour ; EAG ; wind tunnel tests ; field tests ; (Z)-11-hexadecen-1-yl acetate ; (Z)-11-hexadecen-1-ol ; (Z)-11-hexadecenal ; dodecyl acetate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The electrophysiological and behavioural responses of male Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae) to several synthetic blends of its sex pheromone components were investigated. Electroantennogram (EAG) recordings showed the greatest response to a 77:8:10:5 blend of the four compounds found in the female glands: (Z)-11-hexadecen-1-yl acetate, (Z)-11-hexadecen-1-ol, (Z)-11-hexadecenal and dodecyl acetate. A logistic equation was fitted to the dose-response data and differences in relative EAG responses were analysed by deriving five parameters of this equation. Flight responses of males were also evaluated in a wind tunnel where the proposed blend (77:8:10:5) also elicited the best response. Field tests confirmed the laboratory findings as the 77:8:10:5 blend captured more S. nonagrioides males than other pheromone mixtures and showed more selectivity for male S. nonagrioides over other noctuid species, Discestra trifolii Rottenberg and Lacanobia oleracea L., but not for Mythimna unipuncta Haworth, a species with a sex pheromone composition almost identical to that of S. nonagrioides.
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  • 132
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    Entomologia experimentalis et applicata 83 (1997), S. 347-360 
    ISSN: 1570-7458
    Keywords: dispersal ; Lepidoptera ; Mamestra brassicae ; mortality ; Noctuidae ; parasitism ; predation ; survival rate ; white cabbage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mortality of eggs, larvae and pupae and larval dispersal of the cabbage moth, Mamestra brassicae (L.) (Lepidoptera: Noctuidae) was investigated in a series of small-scale field experiments in white cabbage, Brassicae oleracea var. capitata (L.), and in the laboratory during 1990–1992 in south–eastern Norway. The highest mortality was found in young larvae and in hibernating pupae. In 1990, larval mortality in the first instar was 80% (range 9–97% for the individual cohorts). Most larvae died within the first 1–3 days after hatching. The dispersal activity during these days was high, and failure to establish feeding sites and predation were probably the main mortality causes. Pupal mortality during winter was 90% on average for 1990–1993 (range 81–100%). The main mortality factor was probably unfavourable weather conditions, and indications of cold stress were found. The impact from parasitoids and diseases was generally low. Trichogramma semblidis (Aurivillius) (Trichogrammatidae) was reared from M. brassicae eggs in very low numbers in 1991. Larval parasitism increased from 〈 1% in 1990 to almost 24% in 1992, and was almost totally caused by the braconids Microplitis mediator (Haliday) and Aleiodes (Aleiodes) sp. Predation of frozen larvae on the soil surface was 75% on average (range 63–96%) during 1990–1992 in first instar larvae and decreased gradually with larval age. The consumption rates of Philonthus atratus (Gravenhorst) (Staphylinidae) and the carabids Bembidion tetracolum (Say), Pterostichus melanarius (Illiger) and Harpalus rufipes (Degeer) on M. brassicae eggs and larvae were investigated in non-choice experiments in the laboratory. A preliminary survival model based on estimates of the mortality factors identified in this study is presented.
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  • 133
    ISSN: 1570-7458
    Keywords: Lepidoptera ; Tortricidae ; codling moth ; mating disruption ; Isomate-C ; tree banding ; organic apples ; biological agriculture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An integrated programme of pheromone-mediated mating disruption using Isomate-C®, post-harvest removal of fruit, and trapping overwintering larvae with cardboard tree bands, was used to control codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), in four commercial ‘organic’ apple orchards in Cawston, British Columbia during 1989–1992. One application of 1000 dispensers − 1 on May 1 delivered estimated seasonal totals of 16.6, 16.5 and 19.9 g of E,E-8,10-dodecadien-1-ol [=codlemone] − 1in 1990, 1991 and 1992, respectively, at median rates of 8.4, 8.3, and 13.3 mg · 〈 ha−1 · ha−1 during dusk flight periods of first brood and 5.3, 4.7 and 4.6 mg · − 1· ha−1 in second brood, respectively. Over this 3-year period damage from codling moth at harvest ranged from 0.08 to 2.4%, and averaged 60.7% in these four organic orchards, while damage in five conventional orchards receiving sprays of azinphosmethyl ranged from 0.02 to 1.85%, and averaged 0.5%. Damage in an experimental orchard that was banded only, ranged from 43.5 to 56.7%, and averaged 48.9%. Between 1990 and 1992 cumulative male catches in Pherocon 1-CP wing traps baited with 10 mg of codlemone declined by 52% and densities of overwintering codling moth larvae declined an average of 49.5% in all organic orchards. Overwintering populations in the banded experimental orchard showed an increase of 57.7% during this study period. We conclude that an integrated programme of pheromone-mediated mating disruption, post-harvest fruit removal and tree banding, controls codling moth effectively enough to make organic apple production viable in British Columbia.
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  • 134
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    Entomologia experimentalis et applicata 83 (1997), S. 147-159 
    ISSN: 1570-7458
    Keywords: Lepidoptera ; Tortricidae ; visual stimuli ; tactile stimuli ; generalist
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A bioassay to test the ovipositional responses of the generalist herbivore, Epiphyas postvittana, was developed, and various non-chemical factors that influenced the laying, and number of eggs laid, by females were tested. In choice experiments using various models, surface texture was found to be an important factor influencing ovipositional behaviour, with females preferring to lay on smoother rather than rougher surfaces. Another surface texture feature that influenced oviposition was a raised wax ridge on a smooth surface. The wax ridge stimulated more eggs to be laid within close proximity to the ridge. Observations of females showed that the increased numbers of eggs laid on models with a wax ridge resulted not from increased landing, time spent, or numbers of eggs laid per ovipositional bout, but rather through an increased probability of ovipositing after landing on models with a wax ridge compared to models without the ridge. Spatial features of the models were also found to influence egglaying by females. Females showed a tendency to lay on the upper rather than the lower surface of models, although this appeared to depend upon how stimulatory the upper surface of the model was. Females also laid more eggs on models with a dense array of leaves than on models with a more spread-out array.
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  • 135
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    Entomologia experimentalis et applicata 83 (1997), S. 171-180 
    ISSN: 1570-7458
    Keywords: within-plant heterogeneity ; tent caterpillars ; temperature ; circular statistics ; Lepidoptera ; Yponomeutidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The caterpillars of Yponomeuta mahalebella Latr. (Lepidoptera: Yponomeutidae) are monophagous on Prunus mahaleb (Rosaceae) leaves. Adult females deposit eggs in batches, which determines larval gregarious behaviour. Coupled with gregarious behaviour, caterpillars spin silk tents within they will feed until pupation. Distribution of tents in the field, their effect in microenvironmental larval growth conditions and the consequences for adult body mass and survival of larvae were studied. There was a significant trend for tents to be placed with a southward orientation. Within-plant tent orientation was related to the sun-shade pattern experienced, which was the main effect determining thermal differences between the inside and the outside of the tent. These differences appeared to affect larval growth and survival, with higher survival of larvae and heavier adults emerging from tents oriented to the south-east and east, respectively.
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  • 136
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    Journal of insect conservation 1 (1997), S. 43-62 
    ISSN: 1572-9753
    Keywords: Lepidoptera ; biogeography ; endemism ; conservation ; butterflies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract The addition of species to the European butterfly list since 1983 has resulted in a number of highly significant changes. Most important are the increases in the number and proportion of endemics and of rare species, and a regional excess of species and endemics for southern Europe compared to northern Europe. There is also a surplus of Lycaenidae and Satyridae compared to other families, and an increase in species per genus associated with the reduction in genera. These additions raise two issues. First, the potential conservation load for European butterflies is inflated at species level. This is especially the case for southern Europe, which has disproportionate increases in rare and endemic species, more particularly if rarity and endemism are found to equate with threat of extinction. Second, the inflation in rarity and endemism suggests that there is a trend to promote ever more local populations (races, subspecies) to species. The taxonomic status of species being added to the list, a quarter of which are regarded as doubtful, is increasingly difficult to determine. Consequently, there is a danger that this may call into question the validity and objectivity of taxonomic practices, and of databases dependent on them, used by conservation. Revision of higher and lower butterfly taxa is urgently required.
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  • 137
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    Molecular genetics and genomics 255 (1997), S. 605-610 
    ISSN: 1617-4623
    Keywords: Key words Transposable element ; Excision ; Recombination ; piggyBac ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The terminal DNA sequence requirements for piggyBac transposable element excision were explored using a plasmid-based assay in transfected, cultured insect cells. A donor plasmid containing duplicate 3′piggyBac terminal inverted repeats was constructed that allowed individual nucleotides or groups of nucleotides within one of the 3′ repeats to be mutated. The relative extent of excision using the mutated end versus the wild-type end was then assayed. Removal of even one of the terminal 3′ G nucleotides from the piggyBac inverted repeat, or removal of the dinucleotide AA from the flanking TTAA target site prevents excision of piggyBac at the mutated terminus. Incorporation of an asymmetric TTAC target site at the 3′ end does not prevent excision from the mutated end. Thus, both piggyBac DNA and flanking host DNA appear to play crucial roles in the excision process.
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  • 138
    ISSN: 1573-1561
    Keywords: Lepidoptera ; Lymantriidae ; Orgyia pseudotsugata ; tussock moth ; (Z)6,(Z)9-heneicosadien-11-one ; (Z)6,(E)8-heneicosadien-11-one ; (Z)6,(Z)9-heneicosadien-11-one ; sex pheromone ; synergism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Three candidate sex pheromone components, (Z)6,(Z)9-, (Z)6,(E)8-, and (Z)6,(E)9-heneicosadien-11-one (Z6Z9, Z6E8, and Z6E9) were identified in pheromone gland extracts of female Douglas-fir tussock moths (DFTM), Orgyia pseudotsugata (McDunnough). Their occurrence in subnanogram quantities in extracts and structural conversion during analytical procedures and bioassays complicated chemical identifications. Complete identification required comparative analyses of stereoselectively synthesized and female-produced dienones by coupled gas chromatographic–electroantennographic detection (GC-EAD), high-performance liquid chromatography (HPLC) and coupled GC–mass spectrometry (MS). Determination of the pheromone component was contingent upon an experimental design that minimized structural rearrangement of dienones before and during the field test. In a 40-min field experiment, acetonitrile solutions of each of the above dienones were carried on Dry Ice to traps and were syringed onto cotton release devices below trap lids. In combination with the previously known sex pheromone component of DFTM, (Z)6-heneicosen-11-one (Z6), Z6E8 was the only synergistic dienone and the mixture was highly attractive. Because Z6 by itself attracts seven species of tussock moths (two sympatric with DFTM), a blend of Z6 and Z6E8 may impart specificity to DFTM pheromone communication. In commercial lures, this binary blend may facilitate species-specific, sensitive monitoring and efficacious control by mating disruption of this important forest defoliator.
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  • 139
    ISSN: 1573-1561
    Keywords: Lepidoptera ; Noctuidae ; Agrotis ipsilon ; Agrotis segetum ; hybridization ; pheromone ; diapause ; migration ; sexual isolation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Hybrids were obtained by crossing males of the turnip moth Agrotis segetum, which has a period of arrested development in the larval stage, with females of the migrant and nondiapausing black cutworm, Agrotis ipsilon. No progeny were obtained by crossing females of A. segetum with males of A. ipsilon. Backcrosses were successful only by crossing hybrids with both sexes of A. ipsilon. No larval oligopause occurred when hybrid larvae were reared under conditions inducing arrest in A. segetum (12L:12D, 21°C). Wind tunnel tests showed interattractiveness of F1 hybrids with A. ipsilon but not with A. segetum. Single sensillum recordings of pheromone-sensitive hairs on the antennae of males of the parent species revealed differences in the distribution of sensilla. F1 and F2 hybrids exhibited the same receptor cell types and distribution as A. ipsilon. Gas chromatographic analysis of female pheromone gland extracts revealed that F1 hybrids were also very close to A. ipsilon.
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  • 140
    ISSN: 1573-1561
    Keywords: Lepidoptera ; Noctuidae ; Agrotis segetum ; (Z)-5-decenyl acetate ; pheromone analogs ; structure–activity ; bioisosteres ; single-sensillum recordings ; receptor interaction ; molecular electrostatic potential ; quantum mechanical calculations ; ab initio calculations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A number of analogs of (Z)-5-decenyl acetate, a pheromone component of the turnip moth,Agrotis segetum, in which the acetate group has been replaced by functional groups that may function as bioisosters, have been synthesized and tested using single-cell electrophysiology. The activities have been interpreted in terms of the molecular electrostatic potentials of the polar functional group as calculated byab initio quantum mechanical calculations. It is concluded that both oxygens of the acetate group in (Z)-5-decenyl acetate contribute to the interactions between the pheromone component and its receptor. Furthermore, the results indicate that the crucial interaction between the carbonyl group and the receptor, which is most probably a hydrogen bonding interaction, takes place in a direction pointing away from the hydrocarbon chain of the pheromone component.
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  • 141
    ISSN: 1573-1561
    Keywords: Sex pheromone ; processionary moth ; Thaumetopoea pityocampa ; Lepidoptera ; Thaumetopoeidae ; minor component ; single cell recording ; behavior
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The female sex pheromone of the processionary moth Thaumetopoea pityocampa has been reinvestigated to look for possible minor components. Examination by GC-MS and GC-EAD of the contents of virgin female glands, after stimulation with PBAN (pheromone-biosynthesis-activating neuropeptide), showed that the major component, (Z)-13-hexadecen-11-ynyl acetate (1), appears to be the only pheromone compound present in the gland. Comparison of female attractivity with that of the natural extract and synthetic (Z)-13-hexadecen-11-ynyl acetate showed that this chemical is able to elicit a similar activity to that displayed by virgin females in a wind tunnel. In single cell recording experiments, two specialist receptor cell types were found in the trichoid sensilla. One cell type was tuned to enyne acetate 1. The other one was tuned to (Z,Z)-11,13- hexadecadienal and (Z)-13-hexadecen-11-ynal, the major components of the pheromone blend of other Thaumetopoea spp., and constitutes a further example of interspecific inhibitor receptor cells. Our results show that the processionary moth may not need minor components for successful mate recognition.
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  • 142
    ISSN: 1573-1561
    Keywords: Polyommatus bellargus ; Lepidoptera ; Lycaenidae ; Coronilla varia ; Fabaceae ; flavonoids ; sequestration ; insect–plant interactions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Larvae of the lycaenid butterfly Polyommatus bellargus were reared on leaves of Coronilla varia that are rich in flavone C-glycosides. Six flavonoids including isovitexin and isoorientin, as well as several of their congeners were isolated and identified by spectroscopic means. Comparative HPLC analysis of the host plant and of larvae, pupae, and imagines of P. bellargus indicated selective uptake of isovitexin versus isoorientin derivatives. Isovitexin-2″-O-xyloside was the major flavonoid detected in pupae and in imagines of P. bellargus. Several minor components were tentatively identified as quercetin- and kaempferol-O-glycosides based on their on line-UV spectra and by comparison with known standards. Since leaves of the host plant accumulate exclusively flavones, the flavonol glycosides are considered to be biotransformation products that are formed by the insects and/ or by symbiotic bacteria. Imagines of P. bellargus caught in the wild exhibited similar flavonoid patterns compared to imagines reared in the laboratory. Within the imagines, approximately 80% of all flavonoids are stored in the wings (especially in the orange submarginal lunules), whereas the remaining 20% resides in the bodies. Female butterflies show a significantly higher flavonoid concentration than males. It is suggested that the sequestered flavonoids are involved in visual mate recognition.
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  • 143
    ISSN: 1573-1561
    Keywords: Chiral HPLC ; sex pheromone ; Lepidoptera ; Geometridae ; epoxydiene ; stereochemistry ; Mosher's method
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Resolution of insect pheromonal cis-epoxydiene racemates derived from (Z,Z,Z)-3,6,9-trienes with a C18–C23 chain was examined utilizing chiral HPLC columns, and the result showed that a Chiralpak AS column was suitable to separate enantiomers of the 3,4-epoxides, and a Chiralpak AD column was indispensable for the resolution of the racemic 6,7- and 9,10-epoxides. The absolute configuration of the enantiomers of the 3,4- and 9,10-epoxides separated by HPLC was studied after methanolysis of their epoxy rings. Examination of the 1H NMR data from esters of the methoxyalcohols produced by a modified Mosher's method with (S)- and (R)-α-methoxy-α-(trifluoromethyl)phenylacetic acid indicated that the dextrorotatory parent epoxides with a shorter R t were 3S,4R and 9S,10R isomers and the levorotatory enantiomers having a longer R t possessed 3R,4S and 9R,10S configuration. Field tests with both enantiomers of (Z,Z)-6,9-cis-3,4-epoxynonadecadiene separated by HPLC with the chiral column revealed new specific attraction of geometrid forest defoliators, Pachyerannis obliquaria, to the 3R,4S isomer and Zethenia albonotaria nesiotis to the 3S,4R isomer.
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  • 144
    ISSN: 1573-1561
    Keywords: Turnip moth ; Agrotis segetum ; Lepidoptera ; Noctuidae ; sex pheromone ; genetics ; population variation ; pheromone production ; behavioral response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The genetic basis of the differences in female pheromone blend ratio and male behavioral response in the turnip moth,Agrotis segetum, was examined by crossing individuals derived from Scandinavian and Zimbabwean populations. These two populations differ both in the ratio of the three major female pheromone components,Z5–10:OAc,Z7–12:OAc, andZ9–14: OAc and in the behavioral response of the males in both wind-tunnel and field-trapping assays. The female pheromone blend in this study is treated as the log ngZ5–10:OAc/ngZ7–12:OAc and log ngZ9–14:OAc/ngZ7–12:OAc for statistical analysis. The mean log ngZ5–10:OAc/ngZ7–12:OAc, is under control by a major autosomal factor or factors, but it is unclear what genetic factor or factors may control the mean log ngZ9–14:OAc/ngZ7–12:OAc. Frequency distributions of the proportions of each component show wide individual variation and also suggest control ofZ5–10:OAc andZ7–12:OAc by major autosomal factors, which forZ5–10:OAc may show partial dominance. Analysis of male behavioral response to synthetic blends in the wind tunnel yields inconclusive results, but suggests thatA. segetum may have a broad window of response that reflects the range of individual variation in female blends.
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  • 145
    ISSN: 1573-1561
    Keywords: Urania fulgens ; Lepidoptera ; Uraniidae ; Omphalea diandra ; Euphorbiaceae ; alkaloidal glycosidase inhibitors ; glycosidases ; sequestration ; generalist/specialist herbivores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Generalist herbivores of the neotropical liana Omphalea diandra (Euphorbiaceae) were compared to the specialist herbivore, larvae of the uraniid moth Urania fulgens, with respect to their ability to accumulate the alkaloidal glycosidase inhibitors (AGIs) produced by the plant and the resistance of their digestive glycosidases to inhibition by these AGIs. The generalist herbivores did not accumulate the AGI aglycones 2R,5R-dihydroxymethyl-3R,4R-dihydroxypyrrolidine (DMDP) and 2,6-dideoxy-2,6-imino-D-glycero-L-gulo-heptitol (HNJ) to the levels detected in larvae of U. fulgens, which contained 0.05–0.11% dry weight DMDP and 0.17–0.35% HNJ. Glucosides of DMDP and HNJ that were synthesized by O. diandra were either absent from both the generalist and the specialist herbivores or present at low levels (less than 0.01%), even though HNJ-glucoside was often the most abundant AGI in the foliage. Analyses of the herbivores' feces indicated that failure to accumulate AGIs was due to the compounds being metabolized rather than excreted. The digestive glycosidases of U. fulgens larvae were more resistant to inhibition by AGI aglycones than those of the generalist herbivores. Similarly, sucrose and maltose hydrolysis in two of the generalist lepidopteran herbivores, larvae of Panthiades ballus and Theope virgilius, was more resistant to inhibition by DMDP than in larvae of Spodoptera littoralis, a lepidopteran which does not encounter O. diandra in nature. There was little difference in the susceptibility to AGIs of glycosidases from the generalist coleopteran Rhabdopterus fulvipes, which naturally feeds on O. diandra, compared with the coleopteran Dermestes maculatus, which does not. The glucoside of HNJ was found to be a very potent inhibitor of trehalase activity in all the insects examined. AGIs are considered to reduce the nutritional value of O. diandra to nonadapted herbivores rather than be acutely toxic. Nevertheless, U. fulgens does appear to be unique among Omphalea-feeding insects in its ability to accumulate AGIs, suggesting that it gains some advantage from storing these compounds.
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  • 146
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    Journal of chemical ecology 23 (1997), S. 2437-2444 
    ISSN: 1573-1561
    Keywords: Parasitoid ; Hymenoptera ; Scelionidae ; corn ; Lepidoptera ; Noctuidae ; semiochemicals ; Y-tube olfactometer ; kairomone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The olfactory cues involved in the host-finding behavior of Telenomus busseolae Gahan, a solitary egg parasitoid of the corn stalk borer, Sesamia nonagrioides (Lefebvre), were studied. The female wasp was attracted to odors from calling virgin females of S. nonagrioides in a Y-tube olfactometer. No response was elicited by noncalling virgin female moths and/or mated female moths. A four-component commercially available sex pheromone of the corn borer was also tested, as were the individual components of the mixture. T. busseolae respond to the whole blend [(Z)-11-hexadecenyl acetate, (Z)-11-hexadecenol, (Z)-11-hexadecenal, and dodecyl acetate in the ratio of 8.5:1:1:2]. Z11–16:OH was the only compound that elicited no significant activity. These results suggest that the sex pheromone released by the host female S. nonagrioides is used by T. busseolae as an indirect cue to locate egg masses of this host.
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  • 147
    ISSN: 1573-1561
    Keywords: Depressaria pastinacella ; parsnip webworm ; Pastinaca sativa ; wild parsnip ; lutein ; UV avoidance ; phototoxicity ; antioxidant ; carotenoid ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The parsnip webwormDepressaria pastinacella, acquires a distinct yellow stripe when it consumes the yellow flowers of its principal host plantPastinaca sativa, the wild parsnip. Caterpillars raised on artificial diet lacking host-plant material lack this yellow coloration. By chemical characterization and comparison of caterpillars raised on parsnip flowers and on artificial diet, we were able to determine that lutein, along with smaller amounts of other xanthophylls from the host plant, is selectively sequestered in the fat body. In bioassays designed to measure avoidance of ultraviolet light, caterpillars raised on parsnip flowers or on artificial diet supplemented with lutein were less likely to avoid exposure to ultraviolet light than caterpillars raised on unaugmented artificial diet and thus lacking sequestered carotenoids. The ability to sequester xanthophylls, which are highly effective antioxidants, may confer a selective advantage on these caterpillars, whose apiaceous host plants produce large quantities of furanocoumarins, natural products that are photoactivated by light wavelengths in the ultraviolet region; such sequestered pigments may reduce not only the oxidative stress associated with ultraviolet light and diurnal foraging but also the photooxidative stress associated with ingestion of photoactive furanocoumarins.
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  • 148
    ISSN: 1573-1561
    Keywords: Oviposition stimulants ; Colias erate ; Lepidoptera ; Pieridae ; Trifolium repens ; Leguminosae ; cyanoglucosides ; linamarin ; lotaustralin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Host-plant chemicals stimulating oviposition by a Leguminosae-feeding pierid butterflyColias erate poliographyswere isolated and identified from one of its primary host plants, white clover (Trifolium repens). Females readily deposited eggs in response to methanolic extracts of the plant, and subsequent partition of the extracts with organic solvents revealed that chemical constituents critical for host recognition reside in the water-soluble fraction. Further fractionation of the hydrosoluble fraction by column chromatography led to the separation of an active fraction and two cyanoglucosides, linamarin and lotaustralin. Conspicuous oviposition response was evoked by unidentified polar compound(s), while these cyanoglucosides exerted no stimulatory activity by themselves. However, ovipositing females preferred samples containing either of the two cyanoglucosides. In dual-choice bioassays, significantly more eggs were laid on samples admixed with the cyanoglucosides, suggesting that the cyanoglucosides serve as synergistic oviposition stimulants and could play an important role in host selection.
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  • 149
    ISSN: 1573-1561
    Keywords: Apanteles carpatus ; Tinea pellionella ; Hymenoptera ; Braconidae ; Lepidoptera ; Tineidae ; semiochemicals ; coupled gas chromatographic–electroantennographic detection ; host-habitat location ; tritrophic interactions ; geranylacetone ; nonanal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In Y-tube olfactometer bioassays, adult Apanteles carpatus (Say), were attracted to beaver or rabbit pelts infested with larvae of the casemaking clothes moth (CCM)Tinea pellionella L. Porapak Q-captured volatiles from a CCM-infested beaver pelt were also very attractive, whereas isolated CCM larvae or larval feces were not. Coupled gas chromatographic–electroantennographic detection (GC-EAD) analysis of the Porapak Q volatile extract revealed two compounds that elicited responses by A. carpatus antennae. Coupled GC–mass spectrometry (MS) in electron impact and chemical ionization modes of these compounds indicated, and GC-MS and GC-EAD of authentic standards confirmed, that they were nonanal and geranylacetone. While each compound singly did not attract A. carpatusa 1:1 blend of both compounds was as attractive as the volatile extract. Because these compounds are host habitat-derivedA. carpatus must be a habitat rather than host specialist, responding to kairomonal indicators of localized and specific habitats such as animal hair or feather. The tritrophic interaction between A. carpatusits clothes moth hosts and their animal-derived habitats is similar to the well-studied relationship between parasitoids of insect herbivores and their host plant habitats.
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  • 150
    ISSN: 1573-1561
    Keywords: Lobesia botrana ; Lepidoptera ; Tortricidae ; pheromone concentration ; mating disruption ; electroantennogram ; vineyard ; plume structure ; fluctuations ; plant canopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Spatial and temporal distributions and relative concentrations of the pheromone of the European grape vine moth Lobesia botrana (Lepidoptera: Tortricidae) were measured in mating disruption trials with electroantennograms. Measurements were carried out over several years during the flight season of the two generations of this pest insect. In three consecutive years significantly higher mean relative pheromone concentrations were measured in summer during the flight of the second generation of L. botrana than in spring during the flight of the first generation (P 〈 0.001). The relative pheromone concentrations in 1989 reached a maximum between late July and early August, when the highest mean daily temperatures were registered. In addition to the differences in mean relative pheromone concentrations, the spatial and temporal distributions of the pheromone differed significantly between spring and summer. EAG recordings taken in summer showed high, uniform pheromone concentrations in the treated plots, whereas in spring strong temporal and spatial fluctuations were recorded. In a vineyard defoliated by a hailstorm, the mean relative pheromone concentrations measured in summer were not significantly different from those measured in spring (P 〉 0.05), but were significantly lower than those of a nearby intact vineyard (P 〈 0.001). The results provided additional evidence that foliage is an important parameter determining mean pheromone concentrations and temporal and spatial distribution of pheromone in mating disruption trials.
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  • 151
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    Journal of chemical ecology 23 (1997), S. 1119-1130 
    ISSN: 1573-1561
    Keywords: Lepidoptera ; Lyonetiidae ; Lyonetia prunifoliella ; Perileucoptera coffeella ; Lyonetia clerkella ; Leucoptera malifoliella ; sex pheromone ; synergism ; 10,14-dimethyloctadec-1-ene ; 5,9-dimethyloctadecane ; 5,9-dimethylheptadecane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Three methylated hydrocarbons, 10,14-dimethyloctadec-1-ene (10Me14Me-1-ene-18Hy = 5Me9Me-17-ene-18Hy), 5,9-dimethyloctadecane (5Me9Me-18Hy), and 5,9-dimethylheptadecane (5Me9Me-17Hy), are synergistic sex pheromone components of the leafminer Lyonetia prunifoliella. Compounds extracted from female pheromone glands were identified by coupled gas chromatographic–electroantennographic detection (GC-EAD), and one compound, 10Me14Me-1-ene-18Hy, also by coupled GC–mass spectrometry. In field trapping experiments, 10Me14Me-1-ene-18Hy, 5Me9Me-18Hy, and 5Me9Me-17Hy singly were unattractive to males but in ternary combination attracted numerous male moths. Attractiveness of the three-component blend significantly exceeded that of two-component blends. No attraction of males to pheromone lures without 10Me14Me-1-ene-18Hy indicates that this compound is essential for pheromone communication of L. prunifoliella. Common C-5 and C-9 methyl branches in lyonetiid pheromone hydrocarbons suggest a common biosynthetic pathway; the presence of 5Me9Me-17Hy and 5Me9Me-18Hy in pheromone blends of L. prunifoliella and Leucoptera malifoliella provides evidence for phylogeny of lyonetiid chemical communication. Determination of the stereoisomeric composition is required to completely describe the pheromone blend of L. prunifoliella and to support the hypothesis of phylogenetically related sex pheromones.
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  • 152
    ISSN: 1573-1561
    Keywords: Oviposition stimulants ; Idea leuconoe ; Lepidoptera ; Danaidae ; Parsonsia laevigata ; Apocynaceae ; pyrrolizidine alkaloids ; parsonsianine ; parsonsianidine ; 17-methylparsonsianidine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A giant danaid butterfly, Idea leuconoe, specializes on apocynaceous plants such as Parsonsia laevigata, which has been reported to contain pyrrolizidine alkaloids. Females of I. leuconoe deposited eggs in response to methanolic extract of P. laevigata, and subsequent bioassay-guided fractionation of the extract revealed that phytochemicals crucial for host recognition by ovipositing females are Parsonsia-specific macrocyclic pyrrolizidine alkaloids including parsonsianine, parsonsianidine, and 17-methylparsonsianidine. Parsonine, another P. laevigata pyrrolizidine component with a keto-dihydropyrrolizine moiety that is closely related in structure to male pheromones of the butterfly, and several nonhost pyrrolizidine alkaloids were entirely inactive. We interpret these data as strong evidence for an ancestral association through herbivory between danaid butterflies and pyrrolizidine alkaloids.
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  • 153
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    Journal of chemical ecology 23 (1997), S. 2145-2159 
    ISSN: 1573-1561
    Keywords: Lepidoptera ; Plutellidae ; (Z)-11-hexadecenyl acetate ; (Z)-11-hexadecenal ; Plutella xylostella L. ; diamondback moth ; sex pheromone ; dispenser ; cabbage ; pheromone evaporation rate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have investigated the effects of pheromone dispenser design, pheromone release rate, and the products of Z11–16:Ald decomposition on the attractiveness of the pheromone blend (Z)-11-hexadecenyl acetate, (Z)-11-hexadecenal, and (Z)-11-hexadecen-1-ol) in traps to Plutella xylostella males. Rubber minidispensers (K-50) were shown to have an active exposure time of at least two months and in delta traps to be capable of monitoring a population of P. xylostella throughout the summer in Estonia. Pheromone release rates between 8 and 17 ng/hr are recommended for maximum trap catches. The attractive blend contained 15–35% of Z11–16:Ac. Decomposition products of Z11–16:Ald inhibited the activity of the pheromone blend when more than 50% of the aldehyde had decomposed.
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  • 154
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    Journal of chemical ecology 23 (1997), S. 51-70 
    ISSN: 1573-1561
    Keywords: Pieris rapae ; Lepidoptera ; Pieridae ; wheat germ diet ; feeding deterrent ; habituation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Rejection of wheat germ diet by cabbage-reared Pieris rapae larvae is explained by the fact that the diet contains strong feeding deterrents to the larvae. These deterrents were present mainly in hexane and butanol (BuOH) fractions of diet extracts, but the post-BuOH water fraction also showed some deterrent activity. Although minor diet components such as sorbic acid had a weak deterrent effect, wheat germ was primarily responsible for the activity. Seven compounds isolated from the BuOH extract of wheat germ were deterrent to the larvae. UV spectra suggested that most of these compounds are apigenin-based flavones. Fractionation of the hexane extracts showed that most of the active compounds were methanol-soluble, and HPLC of the MeOH fractions revealed the presence of active compounds that were absent in the BuOH extract of wheat germ, as well as other compounds that were common to both fractions. Acceptance of nasturtium by P. rapae larvae reared on wheat germ diet is explained by a type of “cross habituation” of the larvae to feeding deterrents. When larvae were reared on cabbage leaves treated with chemical fractions from wheat germ diet, they readily accepted nasturtium, which is normally refused by cabbage-reared P. rapae larvae, due to the presence of strong feeding deterrents in the plant. A high percentage of diet-reared larvae also fed on Erysimum cheiranthoides and Iberis amara, which are normally rejected. The chemical constituents encountered in wheat germ can almost completely account for the larval acceptance of nasturtium. The suitability of wheat germ diet for rearing phytophagous insects for behavioral assays must be questioned.
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  • 155
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    Journal of chemical ecology 23 (1997), S. 473-485 
    ISSN: 1573-1561
    Keywords: Leucania anteoclara ; Leucania commoides ; Lepidoptera ; Noctuidae ; sex pheromone ; (Z)-11-hexadecenyl acetate ; (Z)-9-tetradecenyl acetate ; (Z)-11-hexadecenyl aldehyde ; (Z)-11-hexadecenyl alcohol ; seasonal flight period ; abundance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The sex pheromone components of two species of Leucania that occur sympatrically in western Canada were identified in abdomen-tip extracts from calling female moths. (Z)-11-hexadecenyl acetate was the main component and (Z)-9-tetradecenyl acetate the second component in both species. The third component necessary for specific attractancy was (Z)-11-hexadecenyl aldehyde for L. anteoclara and (Z)-11-hexadecenyl alcohol for L. commoides. The third component for each species was an attractant inhibitor when added as a fourth component to the attractant blend for the reciprocal species. The most effective synthetic blend for the attraction of males in the field was Z9–14:Ac/Z11–16:Ac/Z11–16:Ald in a ratio of 1:10:4 for L. anteoclara and Z9–14:Ac/Z11–16:Ac/Z11–16:OH in a ratio of 5:4:1 for L. commoides.
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  • 156
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    Journal of chemical ecology 23 (1997), S. 1833-1850 
    ISSN: 1573-1561
    Keywords: Agrotis segetum ; Lepidoptera ; (Z)-5-decenyl acetate ; (Z)-7-dodecenyl acetate ; (Z)-9-tetradecenyl acetate ; sex pheromone ; individual variation ; repeatability ; effluvia entrainment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Individual variation of the female sex pheromone signal in Agrotis segetum was determined by gas chromatographic analysis. The individuals varied extensively, both in absolute and relative amounts of the three major pheromone components; Z5–10:OAc, Z7–12:OAc and Z9–14:OAc. Within individuals there was a strong correspondence between the sex pheromone gland content and the signal actually emitted. Both pheromone amounts and proportions of components were correlated with effluvia and extracts; with coefficients of 0.55 for total amounts and 0.66 for proportions. In addition, the repeatability of emitted pheromone components by individual females over three calling periods was high, with correlation coefficients between 0.43 and 0.72. Variation among individual females, however, was observed. There were no significant differences between wild-collected and laboratory-reared females in average ratios of the major pheromone components in pheromone gland extracts, only in total amounts. The biological significance of the inter-individual variation among females is discussed in the context of population dynamics and sexual selection.
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  • 157
    ISSN: 1573-1561
    Keywords: Spodoptera exigua ; Persea americana ; avocado ; idioblast cells ; persin ; (12Z,15Z)-1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15-diene ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We isolated and identified (12Z,15Z)-1-acetoxy-2-hydroxy-4-oxoheneicosa-12,15-diene (persin) from avocado as a compound toxic to Spodoptera exigua larvae. Persin was obtained from oil extracted from the specialized idioblast cells of avocado fruit. When tested for activity using neonates in no-choice artificial diet bioassays, persin at concentrations of 200 μg/g inhibited growth; larvae gained only ∼50% of the weight attained by control insects after seven days. At concentrations of 400 μg/g or above, larval growth was reduced by 〉70% as measured by weight gain, and significant mortality occurred. In choice tests, persin at concentrations of 400 μg/g or higher deterred feeding, with significantly more larvae being found on control diet than on persin-treated diet. Possible applications of persin as a natural insecticide are discussed.
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  • 158
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    Journal of chemical ecology 23 (1997), S. 2187-2196 
    ISSN: 1573-1561
    Keywords: Setothosea asigna ; nettle caterpillar ; Limacodidae ; Lepidoptera ; sex pheromone ; (E)-9-dodecenal ; (E)-9,11-dodecadienal ; oil palm ; Elaeis guineensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Gas chromatographic–electroantennographic detection (GC-EAD) analyses of female nettle caterpillar, Setothosea asigna, pheromone gland extracts revealed seven antennally active compounds. Based on their retention indices on three fused silica columns (DB-5, DB-23, and DB-210), these compounds were hypothesized and, through comparative GC, GC-EAD and GC-mass spectrometry with authentic standards, confirmed to be Δ10-undecenal, dodecanal, (E)-9-dodecenal (E9–12:Ald), (Z)-9-dodecenal, (E)-9-dodecen-1-ol, (E)-9,11-dodecadienal (E9,11–12:Ald), and (E)-9,11-dodecadienol. E9–12:Ald and E9,11–12:Ald were most abundant in female S. asigna pheromone extracts. In field trapping experiments in Palembang, Indonesia, synthetic E9–12: Ald and E9,11–12:Ald at a 1:1 ratio, but not singly, attracted S. asigna males. Attractiveness of these two aldehydes could not be enhanced further through the addition of their corresponding alcohols and/or other aldehydic candidate pheromone components. Use of E9–12:Ald and E9,11–12:Ald for pheromone-based monitoring of S. asigna populations will require lure formulations that minimize pheromone degradation by ultraviolet radiation and atmospheric oxidation.
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  • 159
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    Entomologia experimentalis et applicata 85 (1997), S. 169-175 
    ISSN: 1570-7458
    Keywords: Cydia pomonella ; Lepidoptera ; Tortricidae ; flight distance ; dispersal ; flight mill ; heritability ; colonisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cydia pomonella L. (Lepidoptera: Tortricidae) is considered to be rather sedentary, but some individuals undertake flights of several kilometres in the field. This paper investigates the genetic influence on this variability. The flight capacity was measured in the laboratory by a flight mill and its heritability was estimated for two different strains. The laboratory strain was kept for more than 45 generations and the field strain from Embrach (northern Switzerland) was recently collected in the field. The multiple-trait-restricted-maximum-likelihood method was used for the estimation of genetic variances and covariances. A mixed full-sib/half-sib design was applied for the field strain and a full-sib design for the laboratory strain. The heritability of total distance was 0.57 for the field strain and 0.37 for the laboratory strain (both sexes). In addition, a heritability of 0.38 for total distance was estimated by parent-offspring regression for the laboratory strain. All three values were significantly different from zero P〈0.05 and show that there is a significant additive genetic influence on flight capacity. The genetic correlations between total distance and other flight traits (total duration, flight velocity, longest flight) were between 0.84 and 1.00 for both strains and suggest that these traits actually belong to a single one. High genetic correlations were also found between total distance and the morphological traits body weight and wing length for the field strain, whereas a negative correlation was found between total flight distance and body weight for the laboratory strain. This difference between the two strains was interpreted as a possible trade-off between flight capacity and fecundity.
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  • 160
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    Entomologia experimentalis et applicata 84 (1997), S. 207-219 
    ISSN: 1570-7458
    Keywords: behavior ; feeding ; movement ; walking ; endotoxin ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Behavioral responses of individual lightbrown apple moth, Epiphyas postvittana (Walker), neonate larvae to Dipel or the Bacillus thuringiensis delta endotoxins CryIAc and CryIBa incorporated into artificial diets were measured in choice and no-choice treatments. Frass production and the spinning of silken shelters were delayed when larvae were given diets containing Dipel or Bt endotoxins. In choice treatments larvae did not exhibit an initial preference for control diets over diets with Dipel or Bt endotoxins but accumulated on control diets over a period of days, apparently because larvae that first settled on Dipel or endotoxin diets abandoned diets while larvae that fed on control diets remained on diets throughout the experiment. Abandonment of diets containing Dipel or endotoxins was also apparent in no-choice treatments. Larvae exposed to such diets were more likely to be observed walking or at different sites from one observation to the next, and produced frass and silken feeding shelters at multiple sites rather than a single site. The implications of these findings for the use of B. thuringiensis as a commercial spray or for the use of B. thuringiensis endotoxin genes as resistance factors to be expressed in transgenic apples are discussed.
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  • 161
    ISSN: 1615-6102
    Keywords: Antibodies ; Exine ; Immunocytochemistry ; Phenols ; Pollen ; Sporopollenin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two different antibodies against bovine serum albumin (BSA)-p-coumaric acid-conjugates were produced and used to localize phenolic compounds in exines of pollen from different species,p-Coumaric acid (pC) was coupled to BSA either via the carboxy group (BSA-pC) or directly to the aromatic ring system (BSA-azopC). The polyclonal antibodies raised in rabbits were characterized by ELISA with homologous and heterologous antigens using turkey ovalbumin as carrier protein. The results showed that the two immune sera directed against BSA-pC and BSA-azo-pC, respectively, were specific forp-coumaric acid and structurally similar compounds. Only a very poor binding by acetic acid-ovalbumin-conjugates and no binding by turkey ovalbumin was detectable. The antibodies reacted with partially purified pollen walls and with highly purified exines. The intensity of the immune reaction was proved to be dependent upon the pollen source and the preparation of the pollen walls. Using light and electron microscopy, it was shown for the first time that, in the exines ofCucurbita maxima, antibody binding was predominantly observed in the region of the germ pore apertures, the outer foot layers, and in the micro- and macrospines. We conclude from this and other earlier published data that phenols are important structural compounds of sporopollenin.
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  • 162
    ISSN: 1432-0878
    Keywords: Key words: Placental stem villi ; Perivascular contractile sheath ; Molecules of adhesion plaques ; Extracellular matrix molecules ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In previous studies, we have shown that smooth muscle cells and myofibroblast subpopulations of the perivascular stem villous sheath of the human placenta contain focal adhesion plaques and talin immunoreactivity. The close association of these cells to elastic and collagen fibres have led to the assumption of a functional myofibroelastic unit within the perivascular stem villous sheath. Interactions between the extracellular matrix and smooth muscle cells depend on a variety of structural protein assemblies. In the present study, we examined, by immunocytochemistry, whether the molecular assembly of extracellular matrix proteins and molecules of focal adhesions, known to be essential for signal transduction in smooth muscle cells, are also found in smooth muscle cells of the perivascular stem villous sheath of the human placenta. Vascular and extravascular smooth muscle cells were immunoreactive for α-actinin, vinculin, paxillin and tensin, the integrin chains α1 and β1, and the basement membrane components laminin and heparan/-chondroitin sulfate proteoglycan perlecan. pp125FAK did not react. In the extracellular matrix of blood vessel walls and the perivascular stem villous sheath, we found immunoreactivity of fibronectin and collagen types I, VI and undulin (collagen type XIV). From our data we conclude that within the perivascular stem villous sheath, there exists a system of signal transduction molecules, indicating a cross talk between the smooth muscle cells of this sheath and their surrounding extracellular matrix.
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  • 163
    ISSN: 1432-0878
    Keywords: Key words: Quinolinic acid ; Interferon-γ ; Kynurenine ; Electron microscopy ; Immunocytochemistry ; Excitotoxicity ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract.  Quinolinate (QUIN), a tryptophan-derived excitotoxin, was localized ultrastructurally in human peripheral blood monocytes/macrophages (MØ) by immuno-electron microscopy. A combined carbodiimide/glutaraldehyde/paraformaldehyde-based fixation procedure was developed for optimal retention of QUIN in the cell as well as minimal loss of ultrastructure; a silver-enhanced colloidal gold detection system was used for electron-microscopic analysis. Gold particles representing QUIN immunoreactivity were associated with the inner side of the plasma membrane in normal MØ. The number of gold particles increased significantly when QUIN levels were elevated by treatment with its precursor kynurenine, but location of the gold particles remained essentially the same under this condition. Treatment with interferon-γ increased the number of Golgi bodies, vacuoles and pseudopodia, reflecting the activated state of the cell. Significantly increased numbers of gold particles representing QUIN were detectable in approximately the same location as in the case of kynurenine treatment. Combined treatment with kynurenine and interferon-γ maximally increased the number of gold particles at the periphery of the cell. The pseudopodia were intensely stained with gold particles, while they were not detectable in the inner part of the cytoplasm or in any other organelle even under this activated condition. The significance of the specific location of QUIN revealed in the present study and its relation to the release and subsequent actions of QUIN are discussed.
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  • 164
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    Cell & tissue research 288 (1997), S. 449-469 
    ISSN: 1432-0878
    Keywords: Key words: Neuronal ultrastructure ; Catecholamines ; Dopamine ; Immunocytochemistry ; Connections ; Quantitation ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Interrelations of tyrosine-hydroxylase-immunoreactive afferent fibres with neuronal elements were studied in central, basal and intercalated nuclei of the rat amygdaloid complex. Comparison with dopamine-β-hydroxylase-immunoreacted and phenylethanolamine-N-methyltransferase-immunoreacted parallel sections indicated that the tyrosine-hydroxylase immunoreaction labelled preferentially dopaminergic axons. At the elec- tron-microscopic level, the majority of tyrosine-hydroxylase-immunoreactive axons possessed small boutons containing small clear vesicles and contacting dendrites, spines or somata of amygdala neurons, forming mostly symmetric synapses. They were often directly apposed to or in the vicinity of unlabelled terminals synapsing on the same structure. Synaptic density was highest in the central lateral part of the central nucleus. In the central and basal nuclei labelled axons synapsed preferentially on small dendrites and dendritic spines, and on somata of a few neurons. A detailed study of the neuronal ultrastructure showed that innervated somata possessed the differential characteristics displayed by the predominant neuron types in the medial and central lateral central nucleus and resembled the typical projection neurons in the basal nuclei. In the paracapsular intercalated cell groups the majority of neurons possessed intense perisomatic innervation by immunoreactive terminals. The results suggest that tyrosine-hydroxylase-immunoreactive, predominantly dopaminergic amygdaloid afferent fibres preferentially modulate the effect of extrinsic inputs into neurons of the central and basal nuclei, while a nonselective regulation is exerted upon the output of paracapsular intercalated neurons. It is suggested that this innervation pattern may be important for the coordinated integration of extrinsic and intraamygdaloid connections and thus for balanced output of the structure.
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  • 165
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    Cell & tissue research 288 (1997), S. 471-483 
    ISSN: 1432-0878
    Keywords: Key words: Insulin receptor ; Insulin-like growth factor-1 receptor ; Immunocytochemistry ; Beta-endorphin ; Phosphotyrosine ; Pituitary ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Previous investigations have demonstrated that peripheral insulin has a significant influence on brain function and that the interaction of insulin with neuropeptides in neuroendocrine regions may be pivotal for the regulation of body metabolism and energy balance. Since the various levels of interactions are only incompletely known, the focus of the present study has been the adenohypophysis of the rat, in which the presence and localization of insulin receptors and the structurally and functionally closely related insulin-like growth factor-1 (IGF-1) receptor has been investigated by light- and electron-microsopic immunocytochemistry. The two receptors are found on separate subpopulations of secretory cells of the pars distalis with a preponderance of IGF-1 receptors in a postero-lateral portion of large endocrine cells, insulin receptors being more widely dispersed throughout the pars distalis in a population of smaller, irregularly shaped cells. Insulin receptors, but not IGF-1 receptors, are also located in a subpopulation of secretory cells in the intermediate lobe. Phosphotyrosine, a marker for substrates of receptor tyrosine kinases, has been detected in numerous cells throughout the anterior and intermediate lobe, including the cell populations containing insulin or IGF-1 receptors, indicating their ability to transduce biological signals in the pituitary in vivo. Almost 90% of cells containing insulin receptors are also immunoreactive for beta-endorphin. In contrast, IGF-1 receptors are almost exclusively located on cells secreting follicle-stimulating hormone, suggesting a regulatory role of IGF-1 in the pituitary gonadotropin system. The relationship between β-endorphin and insulin receptors provides further evidence for the hypothesis that peripheral insulin acts as a regulatory hormone in the control of body energy homeostasis via various steps of the neuroendocrine axis, including opioid peptides in the hypothalamus and pituitary known to play an important role in the regulation of feeding behaviour.
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  • 166
    ISSN: 1432-0878
    Keywords: Key words: Prohormone convertase ; PC1 ; PC2 ; Pituitary gland ; Corticotrope cell ; Melanotrope cell ; Immunocytochemistry ; Rana catesbeiana ; Buto japonicus ; Xeriopus laevis ; Rana brevipoda ; Buergeria japonica (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Specific antisera against mammalian prohormone convertases PC1 and PC2 have been used to examine, light-immunocytochemically, the distribution of these enzymes in the pituitary gland of five different species of anuran amphibians (Rana catesbeiana, Bufo japonicus formosus, Xenopus laevis, Rana brevipoda porosa, and Buergeria japonica). A differential pattern of immunoreactivity of PC1 and PC2 was found among these species. Only PC1 was found in the corticotrope cells of the pars distalis in R. catesbeiana, B. japonicus formosus, and X. laevis. Only PC2 was observed in these cells in B. japonica, whereas both PC1 and PC2 were present in these cells in R. brevipoda porosa. PC2 immunoreactivity was always observed in melanotrope cells in the pars intermedia of all of the species, but it coexisted with PC1 immunoreactivity only in R. catesbeiana and X. laevis. The nerve fibers and terminals in the pars nervosa in all of the species were intensely immunopositive with both PC1 and PC2 antibodies. Immunoelectron microscopy on B. japonicus formosus and B. japonica, by means of double-labeling with gold particles of different sizes, revealed that almost all the adrenocorticotropin-positive secretory granules in the corticotrope cells and α-melanophore-stimulating-hormone-positive secretory granules in the melanotrope cells were also labeled with either PC1 or PC2 antibodies. This study suggests that PC1 and PC2 are involved in the intracellular proteolytic cleavage of proopiomelanocortin in amphibian pituitary glands, a situation similar to that proposed for mammals.
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  • 167
    ISSN: 1432-0878
    Keywords: Key words: Hypothalamus ; GABA ; Neuropeptide Y ; Immunocytochemistry ; Development ; ontogenetic ; Oncorhynchus mykiss (Teleostei) ; Salmo trutta fario (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The neuronal system of the saccus vasculosus of two species of trout was studied with immunocytochemical methods and carboindocyanine-dye (DiI) tract-tracing. The cerebrospinal-fluid-contacting neurons of the saccus were immunoreactive for gamma-aminobutyric acid (GABA), glutamic acid decarboxylase (GAD), and neuropeptide Y (NPY). Immunostaining of alternate sections of the saccus vasculosus of fry with anti-GAD and anti-NPY indicated that these substances were colocalized. The tractus sacci vasculosi and the neuropil of the nucleus sacci vasculosi were also immunoreactive to these substances. The GABA, GAD, and neuropeptide Y immunoreactivity of the saccus vasculosus system appeared early in trout ontogeny. After applying DiI to various levels of the tractus sacci vasculosi of adult trout, we observed massive bilateral saccular projections to the nucleus sacci vasculosi and could follow the course of the sacco-thalamic tract. This tract extended in the subependymal region of the thalamus rostral to the nucleus sacci vasculosi and split into two small tracts that reached the subhabenular-preoptic region. Sacco-thalamic fibers formed extensive periependymal plexuses along their trajectory. Interestingly, no clear evidence of the existence of a saccopetal system was obtained. On the basis of these results, we postulate that the saccus vasculosus system modulates the function of centers of the posterior tubercle and periventricular thalamus.
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  • 168
    ISSN: 1432-0878
    Keywords: Key words: FMRFamide ; Immunocytochemistry ; Central nervous system ; Lumbricus terrestris (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of FMRFamide-like immunoreactive cell bodies and fibers in the nervous system of the earthworm Lumbricus terrestris has been studied by means of immunocytochemistry. The cerebral ganglion contains 150–170 immunoreactive nerve cells that are organized into six major groups in the rostral and five major groups in the caudal part of the ganglion; 160–180 immunoreactive nerve cells are present in the subesophageal ganglion, and 80–90 in the ventral cord ganglia. Immunoreactive neurons of the subesophageal and the ventral cord ganglia show similar distributions, in that FMRFamide-like immunoreactive cells form a ventromedial and a lateral cell group. Neuropil in all parts of the central nervous system shows intensively stained varicose and non-varicose fibers. Each segmental nerve contains FMRFamide-like immunoreactive fibers that can partly be traced to the two muscle layers of the body wall, and a fine immunoreactive network lies among the muscle fibers. A similar network is found in the wall of the alimentary canal. Immunopositive perikarya and fibers have been detected in the prostomial nerves, in the stomatogastric system. Some epithelial cells of the body wall are also immunopositive. The morphological characteristics and localization of FMRFamide immunoreactive neurons suggest that they may be involved in: (1) central integratory processes; (2) neuromuscular regulation in both the body wall and enteric system; (3) sensory processes.
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  • 169
    ISSN: 1432-0878
    Keywords: Key words: Leu-callatostatins ; Callatostatins ; Allatostatins ; Neuropeptides ; Peptide purification ; Immunocytochemistry ; Myoinhibition ; Codling moth ; Cydia pomonella (Tortricidae: Lepidoptera) (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A neuroendocrine peptide of the Leu-callatostatin family, LPVYNFGL-NH2, has been isolated from tissue extracts of 5th instar larvae of the codling moth, Cydia pomonella (Lepidoptera). It is identical to a peptide previously isolated from the blowfly, Calliphora vomitoria (Diptera). The distribution of this peptide within the tissues of C. pomonella has been mapped by immunocytochemistry using antisera raised against LPVYNFGL-NH2. Midgut endocrine cells contain Leu-callatostatin immunoreactivity, as do several paired Leu-callatostatin neurones in the brain and ventral nerve cord. Within the visceral nervous system, the frontal ganglion contains four Leu-callatostatin neurones. Axons from these cells combine with others originating from neurones in the brain and project within the nervi cardiostomatogastrici to innervate the tissues of the foregut. In particular, the oesophageal valve has a prominent ring of Leu-callatostatin-immunoreactive fibres. The synthetic peptide, LPVYNFGL-NH2, has a potent reversible inhibitory effect in vitro on all visible forms of spontaneous contractile activity of the foregut, including closure of the oesophageal valve. Complete myoinhibition is observed at peptide concentrations from 10−10 to 10−16 M. These results, in conjunction with the results of similar studies on cockroaches, crickets and flies, suggest that the Leu-callatostatins are a ubiquitous family of insect neuroendocrine peptides with an important role in the control of gut motility.
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  • 170
    ISSN: 1432-0878
    Keywords: Key words: Plasma membrane ; Goblet cell ; Gap junction ; 16-kDa proteolipid ; Lepidoptera ; Larvae ; V-ATPase ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. We previously cloned from Heliothis virescens a 16-kDa protein that is homologous to other ductin sequences. We also reported its immunolocalization with a specific affinity-purified anti-peptide antibody in the midgut and Malpighian tubule of feeding larvae, and concluded that the cloned proteolipid encodes the V-ATPase proton-transporting subunit c from the V0 sector. We now present the immunolocalization of this protein in the midgut during the L4-L5 larval molt and early post-ecdysis into the fifth instar in H. virescens. The results show that the spatial expression of the 16-kDa protein is developmentally regulated. Labeling by anti-peptide antibody varies during the molt in the midgut goblet cell apical plasma membrane and the goblet cell apical valve. Epifluorescence and confocal microscopy revealed strong anti-ductin labeling in areas of cell-to-cell contact during the molt, and during early post-ecdysis into the fifth larval instar. The characteristic labeling pattern observed in areas of cell-to-cell contact is consistent with the claimed involvement of ductins in gap junctions. Conclusive evidence for the presence of the 16-kDa protein in areas of cell-to-cell contact in the midgut of feeding larvae is, however, lacking. V-ATPase regulation during the molt was also investigated by simultaneous immunohistochemistry with an anti-B subunit antiserum, a probe for the V1 sector.
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  • 171
    ISSN: 1573-6857
    Keywords: allozyme frequencies ; genetic drift ; heterozygote deficiency ; Lepidoptera ; Parnassius mnemosyne
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Allozyme polymorphism was studied in 11 Parnassius mnemosyne (Linnaeus, 1758) populations in North-East Hungary. Significant departures from Hardy-Weinberg equilibrium were observed in several cases due to heterozygote deficiency. Genetic variability did not display geographical pattern; the level of genetic differentiation was similar between adjacent populations and between populations originating from different geographical regions. Even a completely isolated population was not differentiated markedly from the others. Thus, genetic drift can be expected as the main evolutionary force acting in the populations.
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  • 172
    ISSN: 1615-6102
    Keywords: Apical growth ; Brefeldin A ; Immunocytochemistry ; Fungus ; Golgi body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Candida albicans, a dimorphic yeast, has the abililty to switch its growth form between budding growth and hyphal growth. Since fungal growth involves secretory processes, spatial control of secretion should play a crucial role in such a morphogenetic transition. Brefeldin A (BFA), an inhibitor of the membrane trafficking system of eukaryotes, increases the occurrence of Golgi-like cisternae in the yeast. In the present study, BFA was used to obtain further insights into the spatial organization of secretory processes in hyphal growth ofC. albicans. BFA completely inhibited the formation and growth of germ tubes at a concentration of 35 μM or higher. Electron microscopy of BFA-untreated germinated cells revealed many vesicles in the apical region and Golgi-like cisternae in the cytoplasm. In cells treated with 35 μM BFA, the vesicles disappeared from the apical region, and, instead, stacked membrane cisternae and membrane-enclosed spherical dense bodies accumulated in the subapical region. These accumulated structures were positive for both polysaccharide staining and immunocytochemical staining with antibodies raised against cell surface antigens ofC. albicans, as were Golgi cisternae in BFA-untreated cells. In cells treated with a higher concentration of BFA (140 μM), the structures that appeared in cells treated with 35 μM BFA were no longer observed and the endoplasmic reticulum was extended and positive for polysaccharide staining. These results suggested that BFA affects different steps of membrane trafficking in a concentration-dependent manner. The accumulated structures induced by 35 μM BFA seemed to be the altered forms of Golgi cisternae. Their accumulation in the subapical region of the germ tube might indicate that the step(s) in membrane trafficking that are associated with the Golgi pathway are vectorially organized in hyphal growth ofC. albicans.
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  • 173
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    Protoplasma 198 (1997), S. 130-134 
    ISSN: 1615-6102
    Keywords: Casuarina ; Frankia ; Nodules ; Nitrogen-fixation ; Hemoglobin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Immunocytochemical localisation of hemoglobin on sections ofFrankia-infecledCasuarina glauca nodule tissue confirmed its presence in nitrogen-fixing infected cells. Using colloidal gold as the marker, hemoglobin was shown to be restricted to the cytoplasm and nucleus of infected cells. None was associated with endophyte hyphae or uninfected cells. As infection developed, with its associated thickening and modification of host cell walls, the level of label, and by implication, the level of hemoglobin increased.
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  • 174
    ISSN: 1615-6102
    Keywords: Electron microscopy ; UV-induced cell damage ; Photoreceptor ; Eye ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A technique that allows the specific inactivation of short-wavelength-sensitive photoreceptive cells has been needed for a long time. Such a technique could be useful in studies on the role(s) of UV-receptors in circadian rhythmicity, recognition of floral patterns, homing behaviour, and mate selection in arthropods. We provide ultrastructural evidence that short-wavelength receptor cells can be selectively damaged without affecting other spectral-cell types. Since the method does not require the killing of the experimental animal, the latter can be used in behavioural or other follow-up tests.
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  • 175
    ISSN: 1432-0878
    Keywords: Key words Estrogen receptor ; Progesterone receptor ; Gonadotropin-releasing hormone ; Preoptic area ; Hypothalamus ; Immunocytochemistry ; Mink (Mustela vison)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The distribution of gonadal steroid (estrogen, progesterone) receptors in the brain of the adult female mink was mapped by immunocytochemistry. Using a monoclonal rat antibody raised against human estrogen receptor (ER), the most dense collections of ER-immunoreactive (IR) cells were found in the preoptic/anterior hypothalamic area, the mediobasal hypothalamus (arcuate and ventromedial nuclei), and the limbic nuclei (amygdala, bed nucleus of the stria terminalis, lateral septum). Immunoreactivity was mainly observed in the cell nucleus and a marked heterogeneity of staining appeared from one region to another. A monoclonal mouse antibody raised against rabbit uterine progesterone receptor (PR) was used to identify the PR-IR cells in the preoptic/anterior hypothalamic area and the mediobasal hypothalamus (arcuate and ventromedial nuclei). This study also focused on the relationship between cells containing sex-steroid receptors and gonadotropin-releasing hormone (GnRH) neurons on the same sections of the mink brain using a sequential double-staining immunocytochemistry procedure. Although preoptic and hypothalamic GnRH neurons were frequently in close proximity to perikarya containing ER or PR, they did not themselves possess receptor immunoreactivity. The present study provides neuroanatomical evidence that GnRH cells are not the major direct targets for gonadal steroids and confirms for the first time in mustelids the results previously obtained in other mammalian species.
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  • 176
    ISSN: 1432-0878
    Keywords: Key words Immunochemistry ; Chromatography ; Immunocytochemistry ; PACAP ; VIP ; CGRP ; NOS ; GRP ; Gastointestinal tract ; Capsaicin ; Denervation ; Rat (Sprague Dawley ; Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The expression of pituitary adenylate cyclase activating polypeptide (PACAP) was studied in the gastrointestinal tract (GI-tract) of normal rats using radioimmunoassay, chromatography, immunocytochemistry, and in situ hybridization. PACAP-38, PACAP-27, and PACAP-related peptide were demonstrated in all parts of the GI-tract, PACAP-38 being the predominant form confirmed by chromatography. PACAP-immunoreactive nerve fibers and nerve cell bodies were found in the myenteric ganglia throughout the GI-tract. PACAP-containing nerve cell bodies were also demonstrated in the submucous ganglia of the small and large intestine. The synthesis of PACAP in intrinsic neurons was confirmed by in situ hybridization. Double immunostaining showed that PACAP is present in calcitonin gene-related peptide-containing sensory nerve fibers as well as in vasoactive intestinal polypeptide (VIP)- or VIP/gastrin-releasing peptide (GRP)-containing (intramural) nerve fibers in the upper GI-tract and in anally projecting, intrinsic VIP-and VIP/nitric oxide syntase-containing nerve cell bodies and nerve fibers in the small and large intestine. Neonatal treatment with capsaicin significantly reduced the concentration of PACAP-38 in the esophagus, stomach, and colon. Extrinsic denervation decreased the PACAP-38 concentration in the stomach, while no change was observed in the small intestine. These results indicate that PACAP- immunoreactive nerve fibers in the GI-tract originate from both intrinsic (enteric) and extrinsic (presumably sensory) sources suggesting that PACAP may have diverse gastrointestinal functions.
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  • 177
    ISSN: 1432-0878
    Keywords: Key words: Oxytocin ; Immunocytochemistry ; Paraventricular nucleus ; Superior cervical ganglion ; Spinal cord ; Sympathetic nervous system ; Retrograde tracing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The paraventricular nucleus of the hypothalamus is a major integrative nucleus for relaying information from the suprachiasmatic nucleus to the autonomic system. The precise pathway by which this information can influence autonomic functions, such as melatonin synthesis in the pineal gland, is not clear. In the present study, we used a retrograde tracer injected in the superior cervical ganglion to identify spinal preganglionic neurons. One of the main neurotransmitters present in descending projections of the paraventricular nucleus of the hypothalamus, oxytocin, was detected with immunocytochemistry to visualise possible contacts with the neurons located in the intermediolateral column of the spinal cord and projecting to the superior cervical ganglion. Although many appositions could be seen at the light-microscopic level, this abundance could not be confirmed at the electron-microscopic level. The implications of these observations for the overall timing message received by the spinal preganglionic neurons are discussed.
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  • 178
    ISSN: 1432-0878
    Keywords: Key words: NPY ; VIP ; Pelvic ganglia ; Immunocytochemistry ; In situ hybridization ; Retrograde tracing (Fluoro-Gold) ; Genital tract ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The present study investigates the distribution and coexpression of neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) in neurones of the accessory ganglion (AG), hypogastric plexus (HP) and paracervical ganglion (PCG), which compose the pelvic plexus in the female rat. Nerve cell bodies immunoreactive for NPY and VIP represent 84% and 46% of the neurone population in the PCG, respectively, while immunoreactivity for each peptide is observed in about 90% of the AG and HP neurones. Adjacent sections immunostained for NPY and VIP, as well as the use of immunocytochemistry combined with in situ hybridization show that 92% of the VIP-containing neurones in the pelvic plexus also contain NPY. In addition, a retrograde tracing study performed in combination with immunocytochemistry demonstrates that pelvic plexus neurones project preferentially to the lower part of the uterus and to the cervix, and that about 95% of these projecting neurones contain VIP. Taken together, our findings indicate that in the female rat, neurones of the pelvic plexus projecting to the lower genital tract mainly coexpress VIP and NPY, and supply nerve fibres to the vascular and nonvascular smooth muscle in the uterocervical region. Since NPY and VIP exert distinct effects according to the target tissue, our results suggest that neurones coexpressing these peptides play important roles in the local regulation of the vascular bed and motor activity of the lower genital tract.
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  • 179
    ISSN: 1432-0878
    Keywords: Key words: Arginine ; Analogues ; Immunocytochemistry ; Nitric oxide ; Pituitary ; Retina ; Transport ; Rat (Wistar) ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Nitric oxide is formed by the action of nitric oxide synthase upon l-arginine. The efficacy of some exogenously applied arginine analogues in inhibiting nitric oxide synthase and thus nitrergic transmission indicates that neurons producing nitric oxide may possess an arginine transport system. To investigate whether arginine analogues are preferentially transported into nitric oxide-utilising cells or into cells making other neurochemicals, we have raised highly specific antisera against a number of arginine analogues including NG-methyl arginine, d-arginine, NGnitro-l-arginine, NG-nitro-l-arginine methyl ester and canavanine. Retinae were incubated in physiological media containing these analogues and rats were given intraperitoneal injections of the analogues to study the pituitary. Immunocytochemistry and NADPH-diaphorase histochemistry revealed that many of these analogues could be transported preferentially, but not exclusively, into nitric oxide-generating cells. However, some nitric oxide-producing cells apparently lacked the ability to take up some arginine analogues. We conclude that nitric oxide-generating cells in the retina and pituitary possess one or more arginine transporters. Other subsets of neurons that use GABA or glutamate as a neurotransmitter may also accumulate arginine analogues, possibly as a substrate for formation of these neurochemicals.
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  • 180
    ISSN: 1432-0878
    Keywords: Key words: Parietal eye ; Pineal organ ; Retina ; Glutamate ; Aspartate ; Immunocytochemistry ; Electron microscopy ; Lacerta muralis ; Lacerta agilis ; Lacerta viridis (Lacertilia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The fine structure of the organ and the localization of the excitatory amino acids glutamate and aspartate were studied in the parietal eye of lizards by postembedding immunoelectron microscopy. The parietal eye contains cone photoreceptor cells, secondary neurons, and ependymal and lens cells. The photoreceptors form long inner and outer segments, some of them being paired as ”twin-photoreceptors” by zonulae adherentes. Perikarya of neurons bear sensory cilia (containing 9×2+0 pairs of tubules) extending into the intercellular space. No neurohormonal terminals are present in the parietal eye. A higher immunoreactivity to glutamate than to aspartate is found in the photoreceptors and in the secondary neurons of the parietal eye. Glutamate immunogold labeling is more intense in the axonal processes of photoreceptors and neurons and in most of the nerve fibers of the parietal nerve running to the brain stem. Weak aspartate and glutamate immunoreactivity can be detected in the ependymal and lens cells. A similar distribution of immunoreactive amino acids is found in the photoreceptors, secondary neurons, and ependymal glial elements of the pineal organ, and retina of the lateral eye of the same animals. Immunoreactive glutamate accumulates in the axons of photoreceptors and secondary neurons of the parietal eye suggesting that this excitatory amino acid acts as a synaptic mediator in the neural efferentation of the organ. Thus, the efferent light-conducting pathway of the parietal organ is similar to that of the pineal organ and lateral eye retina. As the Mullerian cells of the retina, the ependymal and lens cells of the parietal eye and the ependymal-glial cells of the pineal organ may play a role in the metabolism and/or elimination of excitatory amino acids released by photoreceptors.
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  • 181
    ISSN: 1432-0878
    Keywords: Key words: Apoptosis ; Neuronal plasticity ; Hypothalamus ; Astrocytes ; Immunocytochemistry ; Western blot ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A polyclonal antibody raised against a peptide corresponding to the (2–19) amino-terminal sequence of the Bcl-xL/S protein was used to localize Bcl-x immunostaining in the central nervous system of rats at various postnatal ages. Whereas Bcl-x immunostaining was present in virtually all neurons of young animals (4 days postnatal), this staining became progressively restricted during the course of postnatal development. In adults, Bcl-x immunostaining was particularly strong in certain neurons present in a few hypothalamic nuclei, such as the supraoptic or the arcuate nuclei. Moderate staining was observed in some discrete brain regions, such as the olfactory bulb, the hippocampus, some catecholaminergic nuclei of the brainstem, and the cerebellum. Strong Bcl-x immunostaining was also exhibited in axon-like fibers located in the pyriform cortex, the median eminence, the dorsal medulla oblongata, and spinal cord. Bcl-x immunostaining was also present in astrocytes scattered throughout the white matter in the brain and the spinal cord, but was absent from those located in gray matter. Staining was particularly strongly expressed in reactive astrocytes densely packed along the borders of a central lesion or surrounding them, and in a large number of reactive astrocytes detected at a distance from the lesion. Our data suggest that, in addition to the possible stimulating effects on cell survival generally ascribed to Bcl-x, its maintained expression throughout adulthood or its re-expression following injury characterizes those neuronal or non-neuronal cells of the adult central nervous system that synthesize a range of molecules enabling them to adapt rapidly and successfully to a changing environment.
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  • 182
    ISSN: 1432-0878
    Keywords: Key words: Pineal organ ; Pinealocytes ; Pineal photoreceptors ; Sensory structures ; Photopigment ; Pinopsin ; Immunocytochemistry ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Pinopsin is a photoreceptive molecule cloned from the chicken pineal organ. An antibody highly specific for pinopsin was applied in light- and electron-microscopic immunocytochemical studies of the pineal organ of 1 to 2-month-old chickens. Intense immunoreactivity was found in the follicular lumen at the light-microscopic level. In addition, small immunoreactive spherical or fibrous structures were diffusely distributed at the parafollicular aspect of the pineal organ. To identify immunoreactive elements precisely, we used pre-embedding immunoelectron microscopy. These studies revealed immunoreactive outer segments of pinealocytes arranged closely side by side in the follicular lumina. The thin initial portion of the outer segment arose from a basal body located in the inner segment. Immunoreactive pear-shaped outer segments occupied small lumina. Follicular lumina displayed immunonegative arrays of whorl-like lamellar membranes. Occasionally, these immunonegative structures were surrounded by immunoreactive concentric lamellar complexes. In the parafollicular pineal parenchyma, long slender cilium-like structures or enlarged cilia and concentric lamellar arrays showed intense immunoreactivity. All immunoreactive structures observed in this study were considered to represent outer segments of pinealocytes of the chicken pineal organ.
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  • 183
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    Cell & tissue research 289 (1997), S. 265-274 
    ISSN: 1432-0878
    Keywords: Key words: Retina ; Amacrine cells ; Neurotransmitters ; Immunocytochemistry ; Tyrosine hydroxylase ; Dopamine ; Ichthyophis kohtaoensis (Gymnophiona)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ichthyophis kohtaoensis, a member of the limbless Gymnophiona, has a specialized subterranean burrowing mode of life and a predominantly olfactory-guided orientation. The only visually guided behavior seems to be negative phototaxis. As these animals possess extremely small eyes (only 540 μm in diameter in adults), functional investigations of single retinal cells by electrophysiological methods have so far failed. Therefore, the content and distribution of retinal transmitters have been investigated as indications for a functioning sense organ in an animal that is supposed to be blind. In this study, the organization and development of the dopaminergic system have been examined in the retinae of embryonic, larval, and adult I. kohtaoensis, by using an antiserum against tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine synthetic pathway, and an antiserum against dopamine itself. Labeled somata are situated in the inner nuclear layer and in the ganglion cell layer. Dopamine-positive fibers form a dense diffuse plexus, that covers the whole inner plexiform layer, whereas tyrosine hydroxylase-immunoreactive processes show a tendency to arborize in a stratified manner. Tyrosine-hydroxylase-immunolabeled fibers can occasionally be observed in the optic nerve head of larval stages. During ontogenesis and larval development, the distribution of transmitter-expressing cells changes and their number decreases, but no general degeneration of the visual system is detectable. Adult Ichthyophis still have retinal transmitters, indicating that the eyes, although obviously playing a minor role in a subterranean ecological niche, retain all the elements of functioning sense organs.
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  • 184
    ISSN: 1432-0878
    Keywords: Key words: Intracardiac neurones ; Innervation ; Heart ; Neuropeptide Y ; Immunocytochemistry ; Electron microscopy ; Rat (Sprague Dawley) ; Guinea-pig (Dunkin Hartley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Intrinsic neuropeptide Y-containing neurones in rat and guinea-pig hearts were studied at the ultrastructural level by the pre-embedding peroxidase-antiperoxidase immunocytochemical technique. Intracardiac neuronal cell bodies were often weakly or moderately immunostained, and the labelling was usually pronounced in the Golgi complex, multivesicular bodies, some cisterns of granular endoplasmic reticulum and large granular vesicles. Neuropeptide Y-immunoreactive nerve fibres were also observed in association with intracardiac neurones. A subpopulation of neuropeptide Y-immunoreactive granule-containing cells in the rat heart are described for the first time and were very heavily labelled; other granule-containing cells were non-immunoreactive, but were contacted by neuropeptide Y-containing nerves. Preterminal regions of nerve fibres that were located in nerve bundles were only weakly neuropeptide Y-immunoreactive, in contrast to the heavy labelling observed in varicosities that contained many synaptic vesicles. Many neuropeptide Y-immunoreactive nerve fibres were associated with the coronary vasculature and were particularly prominent in the walls of small arteries and arterioles where labelled nerve varicosities were present close to the smooth muscle cells. Immunoreactive nerves were also seen in the myocardium, usually near to capillaries. In axonal varicosities, the central core of large granular vesicles was immunolabelled, and electron-dense immunoreactive material outlined the membranes of small and large clear vesicles. The significance of neuropeptide Y-immunoreactive intracardiac neurones and granule-containing cells and the origin of associated labelled nerve fibres in the heart are discussed.
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  • 185
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    Biodiversity and conservation 6 (1997), S. 817-836 
    ISSN: 1572-9710
    Keywords: Lepidoptera ; host plants ; cerrado ; Brazil ; diversity ; herbivores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cerrado biome is rich in caterpillar species although the general biology and ecology of most tropical species remains unknown. Three host plant genera (Byrsonima, Erythroxylum and Qualea) were examined for caterpillars in four cerrado sensu stricto areas near Brasi´lia (DF, Brazil), from 1991 to 1995. Altogether, more than 16000 host plants were sampled and less than 20% of them possessed caterpillars. All the caterpillars found were reared under laboratory conditions. We successfully reared 137 species of 24 lepidopteran families. The average number of caterpillar species per host species was 28.3. The faunal similarity among plant genera, as well as among study sites, was low, and not related to the distances between them. The low proportion of host plants with caterpillars and the high incidence of rare species appears to be a general pattern for the cerrado. A large proportion (74%) of the caterpillar species occurred on only one host plant family. The most frequent species were either specialists (restricted to one host plant genus) or generalists. We provide a brief outline of some taxonomic problems, some observations on general biology, and temporal and local patterns of some specific caterpillars species. The methodology used, which included weekly data collection, in restricted study areas of the same habitat, with the same sampling method for recording the caterpillar species on the same host plant species, can be used as a tool to explore biodiversity and to discuss some aspects of the conservation of cerrado insects.
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  • 186
    ISSN: 1572-9788
    Keywords: GNA ; insect resistance ; lectins ; Lepidoptera ; potato ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Insecticidal effects of three plant-derived genes, those encoding snowdrop lectin (GNA), bean (Phaseolus vulgaris) chitinase (BCH) and wheat α-amylase (WAI), were investigated and compared with effects of the cowpea trypsin inhibitor gene (CpTI). Transgenic potato plants containing each of the three genes singly, and in pairwise combinations were produced. All the introduced genes were driven by the CaMV 35S promoter; expression was readily detectable at the RNA level in transformants, but not detectable accumulation of WAI could be detected in transgenic potatoes containing its encoding gene. GNA and BCH were accumulated at levels up to 2.0% of total soluble protein; both proteins were expressed in a functional form, and GNA was shown to undergo 'correct' N-terminal processing. Accumulation levels of individual proteins were higher in plants containing a single foreign gene than in plants containing two foreign genes. Resistance of the transgenic plants to insect attack was assayed by exposing the plants to larvae of the tomato moth, Lacanobia oleracea. All the plants tested which were expressing GNA showed an enhanced level of resistance. Leaf damage was reduced by more than 50% compared to controls; total insect biomass per plant was reduced by 45-65%, but larval survival was only slightly reduced (20%). These results support the hypothesis that GNA has a significant antifeedant effect on insects. Expression of BCH had no protective effect against this insect. Expression of CpTI in transgenic potatoes had similar effects to expression of GNA on total insect biomass and survival, but did not afford protection against insect damage to the plant.
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  • 187
    ISSN: 1432-0878
    Keywords: Key words: Urothelium ; Tartrate-resistant acid phosphatase ; Nitric oxide synthase I ; Superoxide dismutase ; Immunocytochemistry ; Free radicals ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three enzymes, viz., tartrate-resistant acid phosphatase (TRAP), nitric oxide synthase I (NOS-I), and superoxide dismutase (SOD), involved in the production and metabolism of free radicals or radical equivalents, were demonstrated by immunocytochemistry in the urothelium of the ureters of six patients of various ages. Two of these enzymes (TRAP and NOS-I) were colocalized in the most apical and lateral border of the superficial cells of the urothelium. In contrast, SOD showed a patchy or granular distribution within the supranuclear region of these cells. Intra- and subepithelial macrophages exhibited a weak TRAP, but no NOS-I or SOD, immune reaction. On the basis of the immunocytochemical findings, arguments in favor of a cytotoxic function of the superficial cells of the human urothelium are presented.
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  • 188
    ISSN: 1432-0878
    Keywords: Key words: Crustacean hyperglycaemic hormone ; Immunocytochemistry ; Neurohaemal organ ; Daphnia magna ; Artemia salina (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Crustacean hyperglycaemic hormone-immunoreactive neuronal systems are detected in the central and peripheral nervous systems of two entomostracan crustaceans, Daphnia magna and Artemia salina, by immunocytochemistry using specific antisera against crustacean hyperglycaemic hormones of the decapod crustaceans Orconectes limosus and Carcinus maenas. In D. magna, four small putative interneurones are detected in the brain. In the thorax, ten bipolar peripheral neurones are stained by both antisera. They are obviously segmental homologues with centrally projecting axons that form interdigitating varicose fibres and terminals in putative neurohaemal areas next to the surface of the anterior part of the thoracic ganglia. Similar immunopositive neurones occur both in the central and peripheral nervous systems of A. salina. A total of five groups of neurones occur in the protocerebrum, the deutocerebrum and the mandibular ganglion. Some of the protocerebral neurones are bipolar and project to the dorsal frontal organ. A single pair of peripheral multipolar neurones in the maxillary segment projects centrally into the ventral nerve cord and innervates unidentified somatic muscles and tissues in the maxillary and the first appendage segments. None of the brain neurones in both species show similarities to decapod X-organ sinus gland neurosecretory neurones. Chromatography of brain extracts of D. magna combined with immunodot blotting revealed two strongly immunoreactive fractions at retention times close to that of the crustacean hyperglycaemic hormone of crayfish. Moreover, preabsorption controls suggest that the cross-reacting peptides of D. magna and A. salina are structurally closely related to those of decapods.
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  • 189
    ISSN: 1432-0878
    Keywords: Key words: Allatostatin ; Immunocytochemistry ; In situ hybridization ; Hemocytes ; Inhibition of corpora allata ; Diploptera punctata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Allatostatins are neuropeptides that inhibit the production, by the corpora allata, of a major insect hormone, juvenile hormone. These peptides are produced by cells of the brain and ganglia as well as by midgut endocrine cells. Transport from these sites may contribute to the allatostatin content in the hemolymph (insect blood). Using a monoclonal antibody against Diploptera punctata allatostatin I (A-P-S-G-A-Q-R-L-Y-G-F-G-L-NH2) and in situ hybridization with a digoxigenin-labeled cRNA probe generated from a portion of the allatostatin gene, it is demonstrated that allatostatin is present in and synthesized by granular hemocytes of D. punctata. About 5% of the hemocytes react with anti-allatostatin antibody and a similar number hybridize with a cRNA probe that detects allatostatin-specific mRNA. Electron micrographs showed that allatostatin-immunoreactive material occurs in membrane-bound, uniformly dense granules that frequently fill fusiform-shaped cells. Allatostatin in cell and plasma fractions of hemolymph quantified by enzyme-linked immunosorbent assay and by bioassay for inhibition of juvenile hormone synthesis in vitro indicated that about equal quantities (0.1–0.2 fmol/μl) are present in cell and plasma fractions. The production of allatostatin by hemocytes suggests that allatostatins may function as regulatory peptides in hemolymph activities in addition to their other known functions.
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  • 190
    ISSN: 1432-0878
    Keywords: Key words: Aquaporins ; AQPcic ; Filter chamber ; Homopteran insects ; Immunocytochemistry ; Cicadella viridis ; Euscelidius variegatus ; Scaphoideus titanus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In the Homopteran order of insects, the plant xylem feeders exhibit a highly differentiated part of their digestive tract known as the filter chamber. In this tissue, water crosses plasma membranes through a transepithelial osmotic gradient. In previous studies on the filter chamber of Cicadella viridis, we purified and characterized from the plasma membranes a 25 kDa protein that we demonstrated to be an aquaporin (or water channel, member of the major intrinsic protein family, a group of membrane channels for small solutes). We called this protein AQPcic for Cicadella aquaporin. In the present study, we used polyclonal antibody anti-AQPcic in Western blotting and immunocytochemical analysis of the intestinal tract of Cercopis sanguinolenta, Philaenus spumarius, Aphrophora alni (Cercopidae), Euscelidius variegatus, and Scaphoideus titanus (Jassidae). Western blotting experiments revealed that immunologically related AQPcic proteins are found in those species. The molecular weight of these proteins is 15–26 kDa. Immunocytochemical studies on ultrathin filter-chamber sections revealed that the anti-AQPcic antibody systematically labelled the membrane microvilli of epithelial cells. A good correlation thus exists between the physiology of these cells and the presence of aquaporin-related proteins in their membranes.
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  • 191
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    Oecologia 108 (1996), S. 643-651 
    ISSN: 1432-1939
    Keywords: Migration rate ; Lepidoptera ; Mark-recapture study ; Population density ; Movement behaviour
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rapidly increasing fragmentation of natural landscapes decreases the ability of many species to reach the smaller and more isolated patches of habitat in a metapopulation. The densities of local populations of several moth species and the butterfly Hipparchia semele in a network of small islands, and the rates of inter-island movement and movement patterns, were investigated, to determine the factors affecting the rate and pattern of movements. The estimated population densities ranged from 0.001 to 0.2 individuals/m2. The observed emigration and immigration rates depended on island isolation and various traits of the species, with great variability in migration rates among species. Thin-bodied, slow-flying species did not move among the islands, whereas many robust, fast-flying species moved among the islands relatively frequently. Migration rate increased significantly with body size and was significantly higher in oligophagous than in polyphagous species, suggesting that these factors are important determinants of the migration rate of the species. Migration rate was low when the surface temperature of the sea was low, and a greater proportion of individuals emigrated from small than large patches of habitat. The migration distances of female noctuids were shorter than those of males and those of both sexes of the butterfly H. semele. The observed movement patterns are consistent with a metapopulation structure in most of the moth species.
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  • 192
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    Journal of comparative physiology 178 (1996), S. 55-61 
    ISSN: 1432-1351
    Keywords: Electrophysiology ; Lepidoptera ; Photoreceptor ; Spectral sensitivity ; Vision
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract 1. The ommatidia of the butterfly Papilio have a fused and tiered rhabdom. The distal tier of the rhabdom is made up of four distal photoreceptors (R1–4), whereas the proximal tier is made up of four proximal (R5–8) and one basal photoreceptor cell (R9). 2. We first confirmed by light microscopy that the ommatidia of Papilio are not twisted, i.e. have the same spatial organization all about the longitudinal axis. The polarization method, previously applied to the distal tier, hence is applicable to identify the photoreceptor location from the peak angle of the polarization sensitivity. 3. We determined the polarization and spectral sensitivity of in total 109 proximal and basal photoreceptors in the lateral looking eye region. All of the photoreceptors were either green or red type, most of which fall into three classes as judged by the peak angles of the polarization sensitivity: around 40°, 150°, and 180° (= 0°) with respect to the dorso-ventral axis. The first two classes are formed by the proximal photoreceptors with straight microvilli oriented at the average angle of 39° (R6, 8) and 144° (R5, 7) respectively, and the third is formed by the basal photoreceptors R9 with straight microvilli oriented at 180° (= 0°). The mean polarization sensitivity (PS = maximal sensitivity/minimal sensitivity) was about 2. 4. 75% of the proximal and 48% of the basal photoreceptors were of the red type. 5. A single ommatidium of Papilio appears to contain two to four types of spectral receptors.
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  • 193
    ISSN: 1432-1351
    Keywords: Heliothis ; Noctuidae ; Lepidoptera ; Plume structure ; Behavior
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract 1) Male Heliothis virescens moths flew upwind to pulsed pheromone plumes. Upon truncation of the pulsed plume males flew into clean air, turning their tracks crosswind (〉 60° relative to directly upwind direction at 0°) within an average of 0.27 s, and were casting, perpendicular to the wind-line (90°), within 0.43 s. 2) The characteristic casting flight in clean air consisted of left-right crosswind reversals, continuing for many seconds without further pheromonal stimulation. Males intercepting a single strand of pheromone during casting flight responded by surging upwind (track angles 〈 60°). The phasic surge lasted only 0.38 s before reverting to crosswind flight (〉 60°). 3) Average templates of responses in two and three dimensions were created. Males controlled their vertical deviations very tightly when in contact with pheromone but upon entering clean air, lateral and vertical excursions became much greater. 4) Males failed to sustain upwind flight to repetitively pulsed plumes generated at 〈 4 filaments/s. At the threshold frequency of 4 pulses/s we show that upwind flights were composed of reiterated surges followed by crosswind casting. As the pulse frequency increased, the tracks became straighter and the single filament cast-surge-cast template could be viewed only sporadically when, for example, a male apparently failed to intercept filaments.
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  • 194
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    Cellular and molecular life sciences 52 (1996), S. 259-263 
    ISSN: 1420-9071
    Keywords: Senecio cannabifolius ; Compositae ; foliar surface chemistry ; tobacco cutworm ; Spodoptera litura ; Lepidoptera ; Noctuidae growth inhibition ; ethyl (1-hydroxy-4-oxocyclohexa-2,5-dien-1-yl) acetate ; methyl (1-hydroxy-4-oxocyclohexa-2,5-dien-1-yl) acetate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Dewaxed leaf surface extracts of 12 plants from Hokkaido, prepared by dipping fresh leaves in chloroform for 3 min, were used in a choice leaf-disk bioassay against larvae of the tobacco cutwormSpodoptera litura. Activity was found only in the extract ofSenecio cannabifolius, a very successful weed in Hokkaido. Individual fractions of the extract, however, were not active. Incorporation of the individual fractions of the surface extracts as well as fractions of the methanolic extracts of the leaf residue into an artificial diet fed to neonateS. litura led to the isolation of ethyl (1-hydroxy-4-oxocyclohexa-2,5-dien-1-yl) acetate, the major surface compound, as the active principle. This compound was also present in the methanolic extract of the leaf residue together with methyl (1-hydroxy-4-oxocyclohexa-2,5-dien-1-yl) acetate, which had the same growth inhibitory effect on the larvae. The presence of these compounds in the foliar surface and tissue suggests a defensive role against herbivores.
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  • 195
    ISSN: 1420-9071
    Keywords: Mating ; pheromone ; epoxyheneicosadiene ; enantiomer ; Arctiidae ; Callimorpha ; Tyria ; Lepidoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract It has been suggested that a common sex pheromone composition may account for interspecific sexual interactions observed with certain moths in the Arctiidae. In this study, it is demonstrated that the sex pheromones released by females of the Scarlet Tiger Moth,Callimorpha dominula L., and the Cinnabar Moth,Tyria jacobaeae L., have similar activities and elute at the same retention time on analysis by coupled gas chromatography (GC)-electrophysiology with males from each species. Peak enhancement on GC, chiral GC and coupled GC-mass spectrometry using authentic compounds show that the sex pheromone for bothC. dominula andT. jacobaeae is (3Z,6Z,9S,10R)-9,10-epoxyheneicosa-3,6-diene.
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  • 196
    ISSN: 1423-0445
    Keywords: sex pheromone ; synergist ; antagonist ; mate recognition ; reproductive isolation ; chemotaxonomoy ; phylogeny ; evolution ; Lepidoptera ; Tortricidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The geometric isomers (E,E)-, (E,Z)-, (Z,E)-, and (Z,Z)-8,10-dodecadien-1-yl acetate were identified as sex pheromone components or sex attractants in the tribes Eucosmini and Grapholitini of the tortricid subfamily Olethreutinae. Species belonging to the more ancestral Tortricinae were not attracted. Each one isomer was behaviourally active in males ofCydia andGrapholita (Grapholitini), either as main pheromone compound, attraction synergist or attraction inhibitor. Their reciprocal attractive/antagonistic activity in a number of species enables specific communication with these four compounds.Pammene, as well as otherGrapholita andCydia responded to the monoenic 8- or 10-dodecen-1-yl acetates. Of the tribes Olethreutini and Eucosmini,Hedya, Epiblema, Eucosma, andNotocelia trimaculana were also attracted to 8,10-dodecadien-1-yl acetates, but several otherNotocelia to 10,12-tetradecadien-1-yl acetates. The female sex pheromones ofC. fagiglandana, C. pyrivora, C. splendana, Epiblema foenella andNotocelia roborana were identified. (E,E)- and (E,Z)-8,10-dodecadien-1-yl acetate are producedvia a commonE9 desaturation pathway inC. splendana. CallingC. nigricana andC. fagiglandana females are attracted to wingfanning males.
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    Histochemistry and cell biology 106 (1996), S. 9-17 
    ISSN: 1432-119X
    Keywords: Key words Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
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    Histochemistry and cell biology 106 (1996), S. 9-17 
    ISSN: 1432-119X
    Keywords: Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
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  • 199
    ISSN: 1423-0445
    Keywords: protein solubility ; insect yolk protein ; buffering capacity ; phenolics ; Lepidoptera ; gypsy moth ; Lymantria dispar ; Malacosoma disstria ; Choristoneura rosaceana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A standard buffer (5 mM phosphate at pH 7) which is used to extract protein from insect eggs provided complete protein solubility for eggs from three of four tree-feeding lepidopteran species: obliquebanded leaf roller (Choristoneura rosaceana), forest tent caterpillar (Malacosoma disstria), and the eastern tent caterpillar (Malacosoma americanum). Under the same extraction protocol, egg proteins from the gypsy moth (Lymantria dispar), remained nearly insoluble. An array of methods typically used to solubilize insect egg proteins were tried and all but the most denaturing (2% SDS) were ineffective. Extraction buffers with typically high pH values were then evaluated. The results indicated that 1) solubility of gypsy moth egg proteins was pH dependent, and full solubility of most egg proteins required the extraction buffer to have a pH of 12 or more prior to the addition of eggs. We also determined that 2) the gypsy moth egg has a buffering capacity which must be surpassed for complete protein extraction, 3) low salt/high pH buffers gave slightly higher total protein values than did high salt/high pH buffers, 4) parental nutritional history (host species utilized) and egg developmental state (pre-embryonatedvs postembryonated/pre-hatch) were unrelated to the requirements for complete egg protein solubilization, and 5) the presence of soluble phenolics, compounds that have the potential to bind to protein and cause insolubility, was confirmed for the gypsy moth egg with 2-D paper chromatography and several other tests. Based on these results, we present a hypothesis about the cause of egg protein insolubility in the gypsy moth.
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  • 200
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 78 (1996), S. 1-8 
    ISSN: 1570-7458
    Keywords: induction ; imprinting ; learning ; caterpillar ; Lepidoptera ; experience ; diet breadth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This review discusses the ambiguity of results obtained from dual-choice feeding tests, especially in relation to the study of effects of experience. Induced preference experiments done this way can be caused by up to nine potential changes in caterpillar behavior. In addition different physiological mechanisms that could lead to the process of induced preference are discussed. These include habituation to deterrents, the increased ability to process xenobiotics, the acquisition of specific positive responses, sensitization, and associative learning. These different processes might relate to function in different ways so that distinguishing them is important to progress in understanding the phenomenon. Suggestions for experimental designs that provide details on processes involved are indicated.
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