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  • Histochemistry
  • Springer  (17)
  • Nature Publishing Group
  • 1995-1999  (15)
  • 1965-1969  (2)
  • 1935-1939
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  • Springer  (17)
  • Nature Publishing Group
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 4 (1969), S. 260-268 
    ISSN: 1432-0827
    Keywords: Cartilage ; Histochemistry ; Staining ; Protein ; Polysaccharide ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des coupes de cartilage épiphysaire frais de jeunes rats, effectuées à la main, sont colorées à pH=4,5 dans des solutions à 0,01% de divers colorants cationiques, appartenant aux groupes de la thiazine, oxazine, azine, triphénylméthane, acridine, et phthallocyanine. Les granules intracellulaires métachromatiques, mises en évidence antérieurement par le bleu de toluidine, sont également identifiées à l'aide de l'azur A, le bleu de méthylène et le bleu de crésyl. Les granules se colorent moins bien à la thionine, le rouge neutre, la safranine O, le bleu de toluylène et l'acridine orange. Dans les conditions utilisées, la matrice de la zone de réserve et la matrice de la zone hypertrophique inférieure (en voie de calcification) se colorent, alors que les matrices des zones prolifératives et hypertrophiques supérieures ne prennent pas les colorants. La gallocyanine, le violet cristal, la fuchsine basique, l'azocarmin B, le bleu de gallamine et la bleu alcian ne se colorent pas ou donnent des réactions colorées différentes de celles décrites ci-dessus. Il semble que le pK et le poids moléculaire des colorants jouent un rôle important, mais ils ne paraissent pas être les seuls facteurs intervenant dans la coloration des granules. Un changement, lié à la calcification, semble intervenir au niveau du matériel métachromatique (probablement des polysaccharides protéiques), aussi bien dans la matrice que les cellules cartilagineuses épiphysaires.
    Abstract: Zusammenfassung Handpräparierte Schnitte von frischem Epiphysenknorpel junger Ratten wurden bei einem pH von 4,5 in 0,01% igen Lösungen verschiedener kationischer Farbstoffe folgender Klassen gefärbt: Thiazin, Oxazin, Azin, Triphenylmethan, Acridin und Phthalocyanin. Die intracellulären β-und γ-metachromatischen Granula, erstmals mit Toluidinblau im frischen Gewebe nachgewiesen, konnten auch gut mit Azur A, Methylenblau und Brillantkresylblau dargestellt werden. Die Granula konnten ebenfalls, aber weniger gut, mit Thionin, Neutralrot, Safranin D, Toluylenblau und Acridinorange gefärbt werden. Unter diesen Färbungsbedingungen werden die inaktive Matrixzone und die untere hypertrophische (verkalkende) Matrixzone angefärbt, während die proliferative und die obere hypertrophische Matrixzone sich nicht färben. Gallocyanin, Kristallviolett, basisches Fuchsin, Azokarmin B, Gallaminblau und Alzianblau färbten entweder gar nicht, oder gaben ein anderes als das obenbeschriebene Färbemuster. Es wird vorgeschlagen, daß das pK und das Molekulargewicht der Farbstoffe wichtig aber nicht unbedingt die einzigen Faktoren sind, die die Färbung der Granula bestimmen. Die Resultate zeigen, daß eine Veränderung im metachromatischen Material (vermutlich Proteinpolysaccharide) vorliegt, und zwar sowohl in der Matrix als in den Zellen des Epiphysenknorpels; diese Veränderung scheint im Zusammenhang mit der Verkalkung zu stehen.
    Notes: Abstract Hand-cut sections of fresh epiphyseal cartilage from young rats were stained at pH 4.5 in 0.01% solutions of various cationic dyes of the thiazine, oxazine, azine, triphenylmethane, acridine, and phthallocyanin classes. The intracellular β-and γ-metachromatic granules, previously demonstrated in fresh tissues with toluidine blue, were also demonstrated well with azure A, methylene blue, and brilliant cresyl blue. The granules were also demonstrated, but not as well, by thionin, neutral red, safranin O, toluylene blue, and acridine orange. Under the conditions of staining, the reserve zone matrix and the lower hypertrophic (calcifying) zone matrix stained, whereas the proliferative and upper hypertrophic zone matrix did not stain. Gallocyanin, crystal violet, basic fuchsin, azocarmine B, gallamine blue, and alcian blue either did not stain, or gave a different pattern of staining from that described above. It is suggested that the pK and molecular weight of the dyes are important, but not necessarily the only factors in determining the staining of the granules. The results indicate that there is a change in the metachromatic material (presumably proteinpolysaccharide) in both the matrix and cells of epiphyseal cartilage, which appears to be related to calcification.
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  • 2
    ISSN: 1432-041X
    Keywords: Development ; Lectin ; Neoglycoprotein Glycoconjugate ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protein-carbohydrate interactions are supposed to play key roles in the mechanisms of cell adhesion, biosignalling and intracellular routing, warranting the analysis of the developmental course of expression of epitopes of this system. Thus, a panel of carrier-immobilized carbohydrate ligands was used as probes, namely lactose,N-acetylgalactosamine,N-acetylglucosamine, mannose, fucose and maltose. Additionally, an antibody to an endogenous β-galactoside-binding lectin (anti-galectin-1), the biotinylated lectin and two further human lectins, namely the macrophage migration inhibitory factor-binding sarcolectin and serum amyloid P component (SAP) that displays selectivity for sulphated sugars and mannose-6-phosphate, were included. They enabled us to assess the extent of the presence of respective binding sites in fixed sections from human lungs (pulmonary epithelial cells), livers (hepatocytes) and hearts (myocard cells) of 10–50 weeks gestation. Invariably, specific binding was detected in the three organ types, at least in certain stages. In most of the cases, the intensity of staining exhibited developmental regulation. The apparent patterns reveal similarities between the different cell types, as seen with immobilizedN-acetylglucosamine as well as with labelled galectin-1 and sarcolectin. However, drastic differences among such patterns with nearly opposite developmental courses do also occur, as detected for carrier-attached mannose and maltose residues. These results point to a potential importance for the detected glycohistochemical features in human development and substantiate the possibility of differential regulation of the presence of binding sites for distinct sugars within a certain organ and between the individual cell types of the monitored organs.
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  • 3
    ISSN: 1432-0878
    Keywords: Lymphatic vessels ; Mesentery ; Ultrastructure ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die ableitenden Lymphgefäße im Mesenterium des Dünndarms männlicher weißer Ratten wurden elektronenmikroskopisch und histochemisch untersucht. Elektronenmikroskopisch findet sich ein lückenloses Endothel mit reichlich pinocytotischen Vesikeln. Eine Basalmembran ist im Klappenbereich durchgehend, sonst nur bruchstückweise vorhanden. Die darunter gelegene Bindegewebsschicht (Lamina propria interna) ist sehr unterschiedlich ausgeprägt. Die lichtoptische Einteilung in muskelstarke und muskelschwache (klappennahe) Abschnitte des Lymphangions läßt sich elektronenmikroskopisch bestätigen. Die zahlreichen membrannahen Vesikel der Muskelfasern sind bei Kontraktion in zahnartigen Fortsätzen angereichert. Auch beim Normaltier treten sog. „aktivierte“ Muskelzellen auf. Osmiophiles Material ist nach Fettfütterung zwischen den Muskelzellen und in der Lamina propria interna der Gefäßwand vorhanden, nicht dagegen in den Klappen. Der Kontakt zwischen den Muskelzellen erfolgt mittels fingerartiger Fortsätze. Endothel und Klappen sind frei von nervösen Elementen. Nervenendigungen und Axone sind zwischen den Muskelfasern nicht zu beobachten, hingegen ganz selten Ganglienzellen. Histochemisch sind im Endothel starke Aktivitäten der sauren Phosphatase und der Monoaminooxydase nachzuweisen, auch unspezifische Esterasen, ATP-ase sowie verschiedene Dehydrogenasen sind vorhanden. Cholinesterase, Cholindehydrogenase und alkalische Phosphatase fehlen. In der Media sind unspezifische Cholinesterase, unspezifische Esterasen, ATPase, Monoaminooxydase, Succinodehydrogenase und NAD-Diaphorase nachweisbar. Im einzelnen kann die Stärke der Aktivität einem bestimmten Abschnitt (muskelstark/muskelschwach) des Lymphangions zugeordnet werden. Damit ist eine Einteilung der Lymphangione in klappentragende und muskelmanschettenhaltige Anteile auch histochemisch möglich. Im Gegensatz zur Adventitia der Blutgefäße gibt die der Lymphgefäße keine Reaktion auf alkalische Phosphatase. Die dort reichlich vorhandenen Mastzellen sind Naphthol-AS-D-chloracetat-esterase positiv.
    Notes: Summary The large lymphatic vessels in the mesentery of male white rats were investigated with ultrastructural and histochemical methods. Electron micrographs show an uninterrupted simple endothelial layer, the individual cells of which are rich in pinocytotic vesicles. A distinct continous basement membrane is to be found only in the region of the valves, otherwise it is lacking over wide parts of the vessel wall. The so-called lamina propria interna differs greatly in its extent. Electron microscopy confirms the subdivision of the lymphangion into segments rich in muscle cells and others (in the area of the valves) which are less rich in them. Muscle cells contain numerous vesicles in close connection with the cell membrane which in contracted cells are to be found predominantly in toothlike projections. Also in the normal rat, so-called “activated” muscle cells are to be found. After a meal rich in fat, osmiophilic material can be seen between the muscle cells and in the lamina propria interna, however, not in the valves. Muscle cells are in contact with each other by finger-like processes. The endothelial layer and the valves are lacking nervous elements. There are no nerve endings and no axons between the muscle cells, but very rarely ganglion cells can be found. Histochemically, strong activities of acid phosphatase and MAO have been demonstrated in the endothelial layer. Unspecific esterases, ATP-ase and various dehydrogenases are also present, while cholinesterase, cholindehydrogenase and alkaline phosphatase are lacking. In the media, unspecific esterases, unspecific cholinesterase, MAO, succinodehydrogenase and Naddiaphorase can be found. The intensity of enzymatic activities can be correlated to specific segments of the lymphangion. It is, therefore, possible to distinguish by histochemical methods the valve areas, which are poor in muscle cells, from the muscle-cuffs. In contrast to the blood vessels, the adventitia of the large lymphatics is alkaline-phosphatase negative. The mast cells, abundant in the adventitial layer, show α-Naphthol-AS-D-chloroacetate-esterase activity.
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  • 4
    ISSN: 1432-0878
    Keywords: Key words Mechanoreceptors ; Synaptic proteins ; Histochemistry ; Ultrastructure ; Slit sensilla ; Hair sensilla ; Cupiennius salei (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Indirect immunocytochemical tests were used at the light- and electron-microscopic levels to investigate peripheral chemical synapses in identified sensory neurons of two types of cuticular mechanosensors in the spider Cupiennius salei Keys.: (1) in the lyriform slit-sense organ VS-3 (comprising 7–8 cuticular slits, each innervated by 2 bipolar sensory neurons) and (2) in tactile hair sensilla (each supplied with 3 bipolar sensory cells). All these neurons are mechanosensitive. Application of a monoclonal antibody against Drosophila synapsin revealed clear punctate immunofluorescence in whole-mount preparations of both mechanoreceptor types. The size and overall distribution of immunoreactive puncta suggested that these were labeled presynaptic sites. Immunofluorescent puncta were 0.5–6.8 μm long and located 0.5–6.6 μm apart from each other. They were concentrated at the initial axon segments of the sensory neurons, while the somata and the dendritic regions showed fewer puncta. Western blot analysis with the same synapsin antibody against samples of spider sensory hypodermis and against samples from the central nervous system revealed a characteristic doublet band at 72 kDa and 75 kDa, corresponding to the apparent molecular mass of synapsin in Drosophila and in mammals. Conventional transmissionelectron-microscopic staining demonstrated that numerous chemical synapses (with at least 2 vesicle types) were present at these mechanosensory neurons and their surrounding glial sheath. The distribution of these synapses corresponded to our immunofluorescence results.Ultrastructural examination of anti-synapsin-stained neurons confirmed that reaction product was associated with synaptic vesicles. We assume that the peripheral synaptic contacts originate from efferents that could exert a complex modulatory influence on mechanosensory activity.
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  • 5
    ISSN: 1432-0878
    Keywords: Key words: Oogenesis ; Oocytes ; Carbohydrates ; Lectins ; Histochemistry ; Branchiostoma belcheri (Acrania)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of carbohydrate moieties in lancelet (Branchiostoma belcheri) oocytes has been studied at different stages of development, using a peroxidase-labeled lectin incubation technique, the PAS-reaction and Alcian Blue staining. Binding sites of 5 lectins, indicating the presence of different sugar moieties (Wheat germ agglutinin (WGA) for N-acetylglucosamine, Concanavalin A (Con A) for glucose/mannose, Helix pomatia agglutinin (HPA) for N-acetyl-D-galactosamine, Ricinus communis agglutinin (RCA-I) for galactose and Ulex europaeus agglutinin (UEA-I) for fucose), were identified and were shown to undergo considerable variation during oocyte development. In the previtellogenic stage, HPA, RCA-I and UEA-I were not identified on the oocyte surface, but WGA and Con A gave strongly positive reactions at this site. In the cytoplasm, 4 lectins (Con A, HPA, RCA-I and UEA-I) gave a weak or moderate reaction, and Con A was also observed in the perinuclear region. In vitellogenic oocytes, these 4 lectins were found to also bind to the nuclear envelope, karyoplasm and nucleolus, and, with the exception of Con A, could also be found in the nuclei of more mature stages. The cytoplasmic yolk granules and Golgi vesicles of the vitellogenic oocyte, were moderately positive for Con A, HPA, RCA-I and UEA-I, but HPA, RCA-I and UEA-I were only weakly bound at the oocyte surface. In mature oocytes, all 5 lectins bound moderately or strongly to yolk granules and cell surface. HPA, RCA-I and UEA-I bound moderately or strongly to various nuclear compartments. Thus, carbohydrate content varied with the development and maturation of the oocytes, and the PAS results were in agreement with the lectin-binding results. Charged carbohydrate residues were observed in the egg envelope and Golgi bodies. These results suggest that the appearence of Con A-, HPA-, RCA-I- and UEA-I-binding glycoconjugates in the nuclei of developing oocytes show a varying pattern indicating different phases of nuclear activity which correlate with different carbohydrate synthetic activities of the oocyte.
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  • 6
    ISSN: 1432-0878
    Keywords: Hyaluronan ; Hyaluronic acid-binding protein ; Microwaye fixation ; Histochemistry ; Urogenital tract ; Reproductive organs ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The histochemical distribution of hyaluronan was analysed in various urogenital organs of male and female (non-pregnant and pregnant) rats by use of a hyaluronan-binding protein and avidin biotin/peroxidase staining. Microwave-aided fixation was used to preserve the extracellular location of hyaluronan. The concentrations of hyaluronan in the different tissues were measured with a highly sensitive radio-assay. Hyaluronan accumulated predominantly in the connective tissue around smooth muscle fibres and in the subepithelial lamina propria. Abundant hyaluronan also occurred in perivascular and perineural connective tissue. In the female urogenital organs, hyaluronan content was high in the vagina and urinary bladder, and highest in the vagina during pregnancy. In the uterus, the surface epithelium of the endometrium stained intensely. In the ovary, the zona pellucida of the oocyte and the theca interna cell layer of the follicles and the follicular fluid of mature follicles exhibited prominent staining. The corpus luteum was devoid of hyaluronan, whereas enlarged corpora lutea of pregnancy exhibited weak, patchy staining. In male urogenital organs, staining for hyaluronan was absent from the testis and epididymis, whereas the erectile connective tissue of the penis stained intensely. The hyaluronan concentrations were high in penile tissue and urinary bladder, while testis, epididymis and the ductus deferens contained only little hyaluronan.
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  • 7
    ISSN: 1432-0878
    Keywords: Key words: Choline acetyltransferase ; Acetylcholinesterase ; Histochemistry ; Serotonin ; Mechanoreceptors ; Lipofuscin ; Cupiennius salei (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Histochemical and indirect immunocytochemical techniques were used to search for neuroactive substances and transmitter candidates in identified sensory neurons of two types of cuticular mechanoreceptors in the spider Cupiennius salei Keys.: (1) in lyriform slit-sense organ VS-3 (comprising 7-8 cuticular slits each innervated by 2 bipolar neurons), and (2) in tactile hairs (each supplied by 3 bipolar sensory cells). All neurons are mechanosensitive. A polyclonal antibody against choline acetyltransferase (ChAT) strongly labeled all cell bodies and afferent fibers of both mechanoreceptor types. Western blot analysis using the same antibody against samples of spider sensory hypodermis and against samples from the central nervous system demonstrated a clear band at 65 kDa, corresponding to the molecular mass of ChAT in insects. Moreover, staining for acetylcholine esterase (AChE) revealed AChE activity in one neuron of each mechanoreceptor type. Incubation with a polyclonal antibody against histamine clearly labeled one neuron in each set of sensilla, whereas activity in the remaining one or two cells was near background. All mechanoreceptor preparations treated with a polyclonal antiserum against serotonin tested negative, whereas sections through the central nervous system of the same spiders were clearly labeled for serotonin. The presence of ChAT-like immunoreactivity and AChE implicates acetylcholine as a transmitter candidate in the two mechanoreceptive organs. We assume that histamine serves as a mechanosensory co-transmitter in the central nervous system and may also act at peripheral synapses that exist in these sensilla.
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  • 8
    ISSN: 1432-0878
    Keywords: Oogenesis ; Oocytes ; Carbohydrates ; Lectins ; Histochemistry ; Branchiostoma belcheri (Acrania)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of carbohydrate moieties in lancelet (Branchiostoma belcheri) oocytes has been studied at different stages of development, using a peroxidase-labeled lectin incubation technique, the PAS-reaction and Alcian Blue staining. Binding sites of 5 lectins, indicating the presence of different sugar moieties (Wheat germ agglutinin (WGA) for N-acetylglucosamine, Concanavalin A (Con A) for glucose/mannose, Helix pomatia agglutinin (HPA) for N-acetyl-D-galactosamine, Ricinus communis agglutinin (RCA-I) for galactose and Ulex europaeus agglutinin (UEA-I) for fucose), were identified and were shown to undergo considerable variation during oocyte development. In the previtellogenic stage, HPA, RCA-I and UEA-I were not identified on the oocyte surface, but WGA and Con A gave strongly positive reactions at this site. In the cytoplasm, 4 lectins (Con A, HPA, RCA-I and UEA-I) gave a weak or moderate reaction, and Con A was also observed in the perinuclear region. In vitellogenic oocytes, these 4 lectins were found to also bind to the nuclear envelope, karyoplasm and nucleolus, and, with the exception of Con A, could also be found in the nuclei of more mature stages. The cytoplasmic yolk granules and Golgi vesicles of the vitellogenic oocyte, were moderately positive for Con A, HPA, RCA-I and UEA-I, but HPA, RCA-I and UEA-I were only weakly bound at the oocyte surface. In mature oocytes, all 5 lectins bound moderately or strongly to yolk granules and cell surface. HPA, RCA-I and UEA-I bound moderately or strongly to various nuclear compartments. Thus, carbohydrate content varied with the development and maturation of the oocytes, and the PAS results were in agreement with the lectin-binding results. Charged carbohydrate residues were observed in the egg envelope and Golgi bodies. These results suggest that the appearence of Con A-, HPA-, RCA-I- and UEA-I-binding glycoconjugates in the nuclei of developing oocytes show a varying pattern indicating different phases of nuclear activity which correlate with different carbohydrate synthetic activities of the oocyte.
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  • 9
    ISSN: 1432-0878
    Keywords: Ultrastructure ; Muscle growth ; Hypertrophy ; Hyperplasia ; Histochemistry ; Fish ; Sparus aurata, Dicentrarchus labrax (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Fibre-type differentiation of the lateral musculature has been studied in Sparus aurata (L.) and Dicentrarchus labrax (L.) during larval development. Histochemical and ultrastructural techniques show two presumptive muscle layers and two germinative zones of presumptive myoblasts. At hatching, myotomal muscle consists of a monolayer of thin undifferentiated cells near the skin (first germinative zone) overlying another mono-layer of small diameter fibres extending hypaxially and epaxially away from the transverse septum. Below this, there is a much thicker, deep layer of fibres, generally large in diameter and polygonal in shape. The presumptive myoblasts are located between these two layers of fibres in the second germinative zone. Initially, the superficial and deep muscle fibres show high and low myosin ATPase activity, respectively. Both layers grow by generating new fibres from the two mentioned germinative zones. At the end of larval life, the superficial layer changes its histochemical profile from high to low myosin ATPase activity and, at the same time, intermediate or pink muscle fibres can be observed by oxidative activity (the NADH-TR reaction). Morphometric analysis shows a significant increase in mean fibre diameter during successive ages, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibre population from the germinative zones (hyperplastic growth).
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  • 10
    ISSN: 1432-0878
    Keywords: Key words: Choroid plexus ; Anterior choroidal artery ; Nitric oxide synthase ; Immunohistochemistry ; NADPH-diaphorase ; Histochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Nitric oxide (NO) has recently been shown to regulate blood flow to choroid plexus, a specialized brain structure responsible for production of most of cerebrospinal fluid. In the present study, we used a specific polyclonal rabbit antibody against the neuronal isoform of NO synthase (NOS), a synthetic enzyme for NO, to determine the localization of NOS in the choroid plexus of adult male Sprague-Dawley rats. NOS-containing nerve fibers were found in the anterior choroidal artery and its branches, and in stromal blood microvessels. Chronic denervation experiments indicated that these nerve fibers originate predominantly from the sphenopalatine ganglion. NOS-immunopositive staining was also detected in the cytoplasm of choroidal epithelial cells. NADPH-diaphorase, a histochemical marker for NOS, was found to colocalize with NOS-immunoreactive product in both nerve fibers and choroidal epithelium. Both neuronal and epithelium-derived NO may regulate secretory function and hemodynamics of choroidal tissue.
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