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  • Immunocytochemistry  (143)
  • nitrogen
  • Springer  (217)
  • 1995-1999  (135)
  • 1980-1984  (42)
  • 1975-1979  (40)
  • 1925-1929
  • 1995  (135)
  • 1983  (42)
  • 1978  (17)
  • 1977  (18)
  • 1975  (5)
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  • 1995-1999  (135)
  • 1980-1984  (42)
  • 1975-1979  (40)
  • 1925-1929
Year
  • 1
    ISSN: 1570-7458
    Keywords: Brassica rapa ; B. juncea ; consumption rate ; glucosinolates ; growth rate ; indolyl ; isothiocyanate ; Mamestra configurata ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mamestra configurata (Walker) (Lep., Noctuidae) larvae were fed excisedBrassica juncea (commercial brown mustard) orB. rapa cv. Tobin (Canola) foliage of three plant growth stages-rosette (stage 2), stem elongation (stage 3) and flowering (stage 4). Relative consumption rates (RCRi) were not significantly different between the plant species. Within theB. juncea treatments, there were no significant growth stage differences in RCRi. However, withinB. rapa, RCRi increased with advancing plant growth stage. Larvae fedB. juncea foliage had significantly reduced relative growth rates (RGRi) compared to larvae fedB. rapa foliage. Within theB. juncea treatments, RGRi decreased with advancing plant growth stage. There were no significant growth stage differences in RGRi in theB. rapa treatments. RGRi was inversely proportional to the levels of isothiocyanate-releasing glucosinolates in theB. juncea treatments. RCRi was inversely proportional to the levels of indolyl glucosinolates in theB. rapa treatments. Levels of total phenols and catechols inB. juncea did not show any trend which could be related to growth stage effects in the insect nutritional indices. InB. rapa, levels of phenols and catechols in stage 3 and 4 foliage were lower than that of stage 2 foliage. Analyses of total nitrogen in field-grown plants showed reductions in percent nitrogen from rosette to flowering stage foliage. The response ofM. configurata to different growth stages of its host plants are discussed in relation to changing levels of allelochemicals and nitrogen.
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  • 2
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    Entomologia experimentalis et applicata 77 (1995), S. 205-210 
    ISSN: 1570-7458
    Keywords: nitrogen ; maize ; lepidopterous stem and cob borers ; Sesamia calamistis ; Eldana saccharina ; Mussidia nigrivenella ; Cryptophlebia leucotreta ; dead hearts ; stem tunneling ; yields
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of four nitrogen levels (0, 60, 90 and 120 kg N/ha) on growth of maize and development of lepidopterous pests was investigatdd in a field trial. Nitrogen had a positive effect on both plant growth variables (plant height, stem diameter and yield), and development and survival ofSesamia calamistis andEldana saccharina, and thereby increased the incidence of dead hearts and stem tunneling. However, the percent yield loss due to artificial infestation decreased with increasing N application rate from 20% to 11% in the in the 0kg/ha and 120kg/ha treatment, respectively. Using a multiple regression analysis, plant height, plant diameter and stem tunneling were found to be the most important variables explaining the variability in maize yield.
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  • 3
    ISSN: 1572-8927
    Keywords: Supersaturation ; nitrogen ; cosolvent ; cetyltrimethylammonium bromide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The decomposition of ammonium nitrite in water creates a supersaturated solution of nitrogen. The same process occurs in water-organic solvent mixtures. Acetone, dioxane, dimethylsulfoxide (DMSO) and dimethylformamide (DMF) are the cosolvents used in this study. The limits of supersaturation of nitrogen (C SL /mol L−1) were determined in all of these solvent mixtures by releasing the dissolved gas sonicationally and measuring the volume of released gas. C SL was generally increased in the presence of cosolvents. The effectiveness sequence of organic solvents was found to be as DMF〈DMSO〈Dioxanet≅Acetone. Transportation period of small bubbles formed during sonication is changed by compositions of solvent mixtures. These periods may depend on the viscosity of the solution. Effects of the presence of cetyltrimethylammonium bromide were also studied. It was concluded that there may be a relation between the strength of the watercosolvent H-bonds and C SL and all of the measured quantities of this study were generally affected by micelle formation.
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  • 4
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    Journal of solution chemistry 4 (1975), S. 523-534 
    ISSN: 1572-8927
    Keywords: Salting coefficient ; scaled-particle theory ; gas solubilities ; seawater ; helium ; neon ; argon ; oxygen ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Experimental values of the salting coefficients for He, Ne, Ar, O2, and N2 in seawater are compared with values calculated from scaled-particle theory. The agreement is reasonably good; the average difference between calculated and observed values at 25°C is 0.007. Scaled-particle theory predicts correctly thatk s should decrease as the temperature increases and that this effect should be most pronounced at low temperatures. However, the predicted magnitude ofdK s/dt is only about half of that observed.
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  • 5
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
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  • 6
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    Nutrient cycling in agroecosystems 42 (1995), S. 1-11 
    ISSN: 1573-0867
    Keywords: nitrogen ; 15N humic substances ; acid hydrolysis ; analytical pyrolysis ; NMR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Most of the N in surface soils occurs in organic forms. The organic N in soil plays a key role in plant nutrition and soil fertility through its effects on microbial activity and nutrient availability. Typically, about one-third of the fertilizer N applied to temperate-zone soils is immobilized and retained in organic forms at the end of the growing season. A significant portion of this newly immobilized N is no more available to microorganisms and plants than the native humus N. Stabilization processes, probably involving polymerization of amino compounds and polyphenols, result in incorporation of N into humic substances with a concurrent reduction in N availability. This paper presents an account of the forms and nature of organic N in soil, emphasizing possible formation pathways, chemical characterization of humic substances through conventional and solid-state techniques, and the fate and composition of newly immobilized N in soil.
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  • 7
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    Nutrient cycling in agroecosystems 42 (1995), S. 139-148 
    ISSN: 1573-0867
    Keywords: denitrification ; methods ; 15N ; nitrate ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This paper attempts to answer the question: is denitrification a major route of N loss from tropical agro-ecosystems? This question turns out to be very difficult to answer due to a severe shortage of data on this process for tropical agro-ecosystems other than rice. Given this lack of data, I approach this question by analyzing data on denitrification and nitrous oxide flux in tropical native forest and pasture soils and attempt to make some conclusions and pose some hypotheses about the significance of denitrification in tropical agricultural soils. I also briefly review methods for measuring denitrification. The data analysis suggests that denitrification in tropical forest soils is strongly influenced by the nature and amount of soil C and N turnover. Studies to examine differences in denitrification in different tropical agricultural systems should focus on the effects of system management on C and N turnover. The data analysis also suggests that, just as in temperate regions, denitrification may not be a significant route of N loss from most tropical agricultural systems. However, field studies are necessary to determine if this is actually the case.
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  • 8
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    Nutrient cycling in agroecosystems 45 (1995), S. 221-233 
    ISSN: 1573-0867
    Keywords: fertilizer recovery ; modelling ; nitrogen ; nutrient efficiency ; nutrient surplus ; phosphorus ; Poland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Research on nutrient losses from agricultural systems should try to relate these losses to farm characteristics. This was done for private farms in two districts in Poland. Using data from a farm survey, nutrient surpluses and Nutrient Use Efficiency (NUE, defined as the ratio of outgoing and incoming nutrients) were calculated for nitrogen and phosphorus. Both nutrient surplus and NUE are relatively high. A model was developed to estimate surpluses and NUE from farm characteristics like location, farm size, fertilizer application level, animal density, grass production and sugar beet or potato area. The results of the model are satisfying for nutrient surplus (R2=0.9) and nitrogen NUE (R2=0.4). Estimation of phosphorus NUE was not satisfactory. High surpluses are associated with high fertilizer applications, high animal density and high grass production while an increasing share of sugar beets leads to lower surpluses. A high nitrogen NUE is associated with low fertilizer applications, low animal density and little grass production, and with a high sugar beet area share. Results suggest that, with exception of sugar beet, fertilizer recovery in Poland is very low. Sugar beet, however, combines high fertilizer applications with low surpluses and high NUE. The outcome of the model can be used in the design of environmental policies. The paper ends with some remarks on the type of measures that can be taken, and the effects these will have on private farms in Poland.
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  • 9
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    Nutrient cycling in agroecosystems 42 (1995), S. 185-192 
    ISSN: 1573-0867
    Keywords: 15N ; nitrogen ; soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The use of15N as a tracer in soil/plant research is examined. The limitations of the so-called Ndff approach are discussed to show the need to consider not just the fate of the added label but also the path that was followed and the rate of the transformation. The development of15N isotope dilution techniques to determine gross rates of nitrogen transformation in soil is reviewed with some indications as to the further development of the approach.
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  • 10
    ISSN: 1573-0867
    Keywords: Phosphorus ; nitrogen ; iron ; VAM ; Rhizobium ; faba-bean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect of rhizobial inoculation, vesicular arbuscular mycorrhizal (VAM) fungi on the growth, P, N and Fe uptake by faba-bean plants (Vicia faba L.V. Giza 2) grown in virgin sandy soil, treated with super or rock-P were studied under green-house conditions. The earthern pots received a half of the recommended rate of P either as single super phosphate or rock-P in the rate of 20 mg P/kg soil, and calcium ammonium nitrate was added in the rate of 10 mg N/kg soil. Iron was applied in two levels 0 and 5 mg Fe/kg soil, in the form of iron sulphate (FeSO4·7H2O). Dry matter yield, as well as P, N and Fe-uptake were determined. Nodule numbers and their dry weights, spore numbers and mycorrhizal root infection were determined. Results indicated that, fungal infection and rhizobial inoculation either alone or in combination increased dry matter yield as compared to uninoculated plants, whereas the percentages increase in dry matter were 34, 26 and 57% in case of super-P application, while they were 56, 47 and 89% in case of rock-P when inoculated withRhizobium, VAM and dual inoculation respectively. Also P, N and Fe uptake were significantly increased due to inoculation, and dual inoculation resulted the highest effect. Generally, inoculation withRhizobium and/or mycorrhizae can remove the deficient effect of P and Fe on N2-fixation and plant growth in the soil of low nutrients content.
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  • 11
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    Landscape ecology 10 (1995), S. 337-348 
    ISSN: 1572-9761
    Keywords: fire ; nitrogen ; NDVI ; transect ; watershed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transects across watersheds with varying fire histories and remotely-sensed data were used to study vegetation-resource interactions in a tallgrass prairie in Kansas. Paired plots (fertilized, control) were established along these transects and sampled for grass and forb biomass during the 1989 and 1990 growing seasons. Fertilization resulted in significant production responses in grass and total biomass on the west slopes of the annually burned (ID) and infrequently burned (N4) watersheds for both years (p = 0.05). In 1989, fertilization also produced a significant increase in grass biomass on the west slope of the unburned transect (p = 0.05), however, total production was not significantly increased. East slopes were insensitive to nitrogen additions. Differences in production response along these transects were assessed by testing the interaction between fertilization response and slope position. Significant interactions occurred on both 1D and N4, but only in 1990. Production data for both years were also compared to Normalized Difference Vegetation Index (NDVI) values derived from thematic mapper (TM) images for 1989 and 1990. When differences among transects or watersheds were statistically different, a positive relationship between NDVI and biomass was observed. NDVI values accurately reflected the spatial patterns of production along these transects for both years although not necessarily the magnitude.
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  • 12
    ISSN: 1432-041X
    Keywords: Vitellin ; Yolk granule ; Yolk protein ; Silkworm ; Embryogenesis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Vitellin was purified from eggs of the silkworm,Bombyx mori, by a new method in which vitellin was extracted from isolated yolk granules. The purified vitellin had a molecular weight of 540,000. An antibody against purified vitellin was prepared in rabbits. It reacted with the hemolymph vitellogenin as well as with purified vitellin, but not with other proteins in the hemolymph or in the extract from yolk granules. The anti-vitellin IgG was used to immunocytochemically locate vitellin in theBombyx non-diapause egg during early developmental stages. In the egg, just after oviposition, vitellin was located in internal yolk granules and in small yolk granules of the periplasm. During the early developmental stages studied, vitellin was not metabolized uniformly throughout the egg. The vitellin of the internal yolk granules located at the posterior-dorsal part and of the small peripheral yolk granules was utilized in 16 h and 2 days, respectively, after oviposition. A thin, very vitellin-poor layer was located between the periplasm and the vitellin-rich interior in the newly laid egg. it was always in close contact with the periphery where blastoderm and germ-band cells developed.
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  • 13
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    Calcified tissue international 24 (1977), S. 223-229 
    ISSN: 1432-0827
    Keywords: Enamel-cementum-morphology ; Immunocytochemistry ; Biochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The continuously erupting rabbit incisor tooth is normally thought of as having an enamel covered “crown” on its labial surface and a cementum covered “root” on its lingual surface. We have examined both surfaces of continuously erupting rabbit incisor teeth taken from near term embryos by a variety of means, including transmission and scanning electron microscopy, biochemical fractionation, and immunohistochemistry. In all cases, we could detect no qualitative difference in the early extracellular matrices taken from the labial and lingual surfaces of the teeth. Both matrices were shown to be composed of dentin and enamel, although the thickness and geometry of the enamel matrix on the lingual surface was somewhat different from that on the labial surface.
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  • 14
    ISSN: 1432-2048
    Keywords: Avena ; Immunocytochemistry ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytochrome of oat (Avena sativa L., cv. Garry) coleoptile cells in the red-light-absorbing form, Pr, is diffusely distributed while after conversion to the far-red-light-absorbing form, Pfr, it is observed only in very small areas within the cell. Comparison of phytochrome photoversibility measurements to the distribution of the pigment within the cell indicates that the spectral assay is not influenced by the observed compartmentalization of the chromoprotein. However, the observed compartmentalization of phytochrome is correlated with a loss in spectrophotometrically detectable Pr.
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  • 15
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    Plasma chemistry and plasma processing 15 (1995), S. 279-307 
    ISSN: 1572-8986
    Keywords: Transport coefficients ; transport properties ; viscosity ; thermal conductivity ; electrical conductivity ; diffusion coefficient ; Chapman-Etskog method ; air ; argon ; nitrogen ; oxygen ; plasma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Technology
    Notes: Abstract Calculated values of the viscosity, thermal conductivity and electrical conductivity of air and mixtures of air and argon, air and nitrogen, and air and oxygen at high temperatures are presented. In addition, combined ordinary, pressure, and thermal diffusion coefficients are given for the gas mixtures. The calculations, which assione local thermodynamic equilibrium, are performed for atmospheric pressure plasmas in the temperature range from 300 to 30,000 K. The results for air plasmas are compared with those of published theoretical and experimental studies. Significant discrepancies are found with the other theoretical studies; these are attributed to differences in the collision integrals used in calculating the transport coefficients. A number of the collision integrals used here are significantly more accurate than values used previously, resulting in more reliable values of the transport coefficients.
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  • 16
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    Water, air & soil pollution 85 (1995), S. 677-682 
    ISSN: 1573-2932
    Keywords: soils ; acidification ; ancient woodland ; nitrogen ; environmental change
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract This study outlines the results of analysis of soil samples collected from fixed quadrats located in a regular pattern across Wytham Wood. Oxfordshire, England. The site contains plots of mixed deciduous ancient woodland and more recent plantations. A previous soil study was undertaken in 1974 and samples archived. Soils were resampled in 1991 and some re-analysis of the 1974 samples was undertaken. Soils were of a wide range in types from sands to gravels with a pH range of 3.0–7.0. Results showed some decline in pH in lower horizons, but most striking was a large increase in soil nitrogen for all horizons and soil types.
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  • 17
    ISSN: 1573-2932
    Keywords: air pollution ; sulphur ; nitrogen ; base cations ; throughfall ; Scots pine ; needle elements ; soil leachate ; N.-W. Russia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Bulk precipitation and throughfall analyses in 50–100-year-old Scots pine stands revealed decreasing sulphur, nitrogen, calcium and magnesium deposition gradients, which extend from the St. Petersburg-Leningrad region and N.-E. Estonia to S.-E. Finland. The Ca and Mg deposition alleviate the acidifying effect of sulphur and nitrogen. The Scots pine canopies acted as a sink for ammonium and nitrate, while the canopy interactions increased sulphur, calcium and magnesium content in throughfall. Foliar S, N and Ca concentrations correlated positively with the corresponding deposition loads. In contrast, low foliar magnesium concentrations were detected in the vicinity of St. Petersburg. The results indicate that the sulphur and calcium deposition may have increased soil leachate S and Ca concentrations in the most polluted Scots pine stands.
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  • 18
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    Water, air & soil pollution 85 (1995), S. 623-628 
    ISSN: 1573-2932
    Keywords: monitoring ; deposition ; sulphur ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The total deposition of sulphur (S) and nitrogen (N) components in Norway during the period 1988–1992 has been estimated on the basis of measurement data of air- and precipitation chemistry from the national monitoring network. There are large regional variations in depositions with highest values in the southwestern part of Norway. Time series analysis of annual mean concentrations of sulphur dioxide (SO2) and sulphate (SO4 −−) in air, non marine SO4 −−, nitrate (NO3 −) and ammonium (NH4 +) in precipitation, shows a significant reduction in the S concentrations both in air and precipitation. In precipitation the concentrations are reduced by 30–45 percent in Southern Norway and 45–55 percent in Central and Northern Norway. Even larger reductions are observed in air concentrations with 50–65 percent reduction in Southern Norway and 65–88 percent reduction further north. For N components there are generally no significant trends in concentration levels nor in precipitation or air. The observed trends are comparable with reported trends in emission.
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  • 19
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    Water, air & soil pollution 85 (1995), S. 1613-1622 
    ISSN: 1573-2932
    Keywords: Deposition ; ecosystem ; nitrogen ; Norway spruce ; nutrient cycling ; production ; root function ; sulphur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract In this paper we try to interpret results from different investigations where an ecosystem with Norway spruce was manipulated with increased N and S deposition via the soil system. The site, in Skogaby in Southwest Sweden, had 1989–93 an annual deposition of 9 kg NH4-N; 7 kg NO3-N and 20 kg SO4-S ha−1. The stand was treated during 6 years with 100 kg N and 114 kg S ha− y−1 in the form of ammonium sulphate (NS treatment). The stand reacted with increased above ground production of 31% after 3 years of treatment. The uptake above ground of N was 155 kg ha−1 higher than in the control. Those trends were even stronger after 6 years of treatment. There were no decreases in the uptake of P, K, Ca or Mg (but for B) after 3 or 6 years of NS-treatment. Needle macro nutrient concentrations in relation to N decreased for several nutrients due to dilution effects. As result of the NS treatment pH increased markedly in the litter layer, and less, but significantly, in the humus layer. A decrease in pH value by about 0.3 units was found in the rest of the soil profile down to 50 cm. Dry mass of needle litter fall and litter layer both increased as a result of 6 years of NS-treatment. After three years of treatment 77–80% of all living fine roots in both control and NS treatment were found in the humus layer and the upper 10 cm of the mineral soil. The amount of living fine roots in the humus layer of NS-treated trees decreased to about one third of the control, and the amount of dead fine roots increased by 150% compared with untreated trees after 6 years of treatment. It is argued that the decreased amount of living and increased amount of dead fine roots not necessarily are indications of decreased root vitality. It can also be explained by increased root turnover rate and decreased decomposition rates of N rich new and old fine root litter. No inorganic N was leached from the control plots whereas the NS treated plots started to leach NO3 the second year of treatment. During 1989–1993 a total of 44 kg NO3-N and 30 kg NH4-N per ha was lost from the system which means that 88% of the N supplied was retained by the ecosystem. At first SO4 was adsorbed in the soil, but after five years of treatment the output was almost equal to the input.
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  • 20
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    Water, air & soil pollution 85 (1995), S. 15-24 
    ISSN: 1573-2932
    Keywords: Acid deposition ; global change ; Asia ; fertilizer ; nitrogen ; sulfur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Most acid-deposition investigations have been concerned with the impact of nitrogen oxides (NOx) and sulfur dioxide (SO2) emissions on Europe and North America. This paper examines three issues beyond this central focus. Major conclusions are 1) ammonia (NH3) emissions and subsequent nitrogen (N) accumulation in terrestrial ecosystems have the potential to generate significant acidification in terrestrial and aquatic ecosystems; 2) sulfur (S) and N accumulation in environmental reservoirs will not only result in significant and extensive acidification but will also impact the earth's radiation balance, tropospheric oxidizing capacity, ecosystem nutrient balance and groundwater quality; and 3) future emissions will substantially increase in the developing world, especially in Asia. By 2020, Asian emissions of SO2, NOx and NH3 will be equal to or greater than the combined emissions from Europe and North America.
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  • 21
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    Water, air & soil pollution 85 (1995), S. 101-110 
    ISSN: 1573-2932
    Keywords: carbon ; nitrogen ; sulfur ; biogeochemistry ; mitigation ; global change
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Forest systems cover more than 4.1×109 ha of the Earth's land area. The future response and feedbacks of forest systems to atmospheric pollutants and projected climate change may be significant. Boreal, temperate and tropical forest systems play a prominent role in carbon (C), nitrogen (N) and sulfur (S) biogeochemical cycles at regional and global scales. The timing and magnitude of future changes in forest systems will depend on environmental factors such as a changing global climate, an accumulation of CO2 in the atmosphere, and increase global mineralization of nutrients such as N and S. The interactive effects of all these factors on the world's forest regions are complex and not intuitively obvious and are likely to differ among geographic regions. Although the potential effects of some atmospheric pollutants on forest systems have been observed or simulated, large uncertainty exists in our ability to project future forest distribution, composition and productivity under transient or nontransient global climate change scenarios. The potential to manage and adapt forests to future global environmental conditions varies widely among nations. Mitigation practices, such as liming or fertilization to ameliorate excess NOx or SOx or forest management to sequester CO2 are now being applied in selected nations worldwide.
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  • 22
    ISSN: 1573-2932
    Keywords: forest soil ; leaching ; lysimeter ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Three years of N application to a Cambic arenosol (Typic Udorthent) in two lysimeter series, one with and one without young saplings of Pinus sylvestris L. have produced significant changes in soil solution and leachate chemistry. An application of 30 kg N/ha*yr−1 significantly increased NO3 − leaching from the soil. This N load was also sufficient to significantly increase the mobility of the phyto-toxic elements Al3+ and Mn2+, likewise to increase leaching of the important plant nutrients Ca2+, Mg2+ and K+. At a N load of 90 kg N/ha*yr−1 significant increase in NH4 + leaching was observed, but total leaching of NH4 + was still very low compared to NO3 − leaching. No significant treatment effects were found for SO4 2−, Fe2+ and Cl− in the leachate. Trees grown in the lysimeters buffered the acidifying effect of N application and increased the leachate pH by 0.2 pH units compared to lysimeters without trees.
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  • 23
    ISSN: 1573-2932
    Keywords: sitka spruce ; nitrogen ; deposition ; leaching ; proton production ; green spruce aphid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Studies of biogeochemical cycling and soil acidification have been carried out in even aged stands of Norway spruce, sitka spruce, Douglas fir, beech and oak under the frame of “The Element Cycling Project”. Deposition of excess nitrogen to forests is important as a potential acidifying input. In Denmark, reduced vitality in Norway spruce has promoted extensive planting of sitka spruce. However, several spruce aphid infestations have caused defoliation in many sitka spruce stands. The objectives of this study were to evaluate the effects of deposition and increased litterfall due to spruce aphid infestations on nitrogen transformations in the forest floor in sitka spruce stands on different soil types. The deposition of throughfall nitrogen range from 19 to 35 kg/ha/year. Fluxes of nitrogen in litterfall ranged from 21 to 77 kg/ha/year, whereas nitrogen leaching range from 1 to 57 kg/ha/year. Leaching was lowest at the infertile sites, but increased with magnitude of deposition and aphid infestations. Proton production according to the nitrogen transformations was largest at the fertile site most often affected by infestations. Huge amounts of bird droppings, honey dew and input of easily available nutrients by canopy leaching probably induced litter decomposition and formation of NO 3 − in the soil water.
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  • 24
    ISSN: 1573-2932
    Keywords: atmospheric deposition ; high elevation forests ; foliar uptake ; cloudwater ; nitrogen ; sulphur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract From 1986–1989, a team of scientists measured atmospheric concentrations and fluxes in precipitation and throughfall, and modeled dry and cloudwater deposition in a spruce-fir forest of the Great Smoky Mountains National Park which is located in the Southern Appalachian Region of the United States. The work was part of the Integrated Forest Study (IFS) conducted at 12 forests in N. America and Europe. The spruce-fir forest at 1740 m consistently received the highest total deposition rates (∼2200, 1200, and 700 eq ha−1 yr−1 for SO4 2−, NO3 −, and NH4 +). During the summers of 1989 and 1990 we used multiple samplers to measure hydrologie, SO4 2−, and NO3 − fluxes in rain and throughfall events beneath spruce forests above (1940 m) and below (1720 m) cloud base. Throughfall was used to estimate total deposition using relationships determined during the IFS. Although the SO4 2− fluxes increased with elevation by a factor of ∼2 due to higher cloudwater interception at 1940 m, the NO3 − fluxes decreased with elevation by ∼30%. To investigate further, we began year round measurements of fluxes of all major ions in throughfall below spruce-fir forests at 1740 m and at 1920 m in 1993–1994. The fluxes of most ions showed a 10–50% increase with elevation due to the ∼70 cm yr−1 cloudwater input at 1920 m. However, total inorganic nitrogen exhibited a 40% lower flux in throughfall at 1920 m than at 1740 m suggesting either higher dry deposition to trees at 1740 m or much higher canopy uptake of nitrogen by trees at 1920 m. Differential canopy absorption of N by trees at different elevations would have significant consequences for the use of throughfall N fluxes to estimate deposition. We used artificial trees to understand the foliar interactions of N.
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  • 25
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    Water, air & soil pollution 85 (1995), S. 1765-1770 
    ISSN: 1573-2932
    Keywords: nitrogen ; sulphur ; input ; forested catchment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The study covers 1991–1994 concentrations of SO2 and NO2 in the air, concentrations of sulphur and nitrogen in bulk precipitation, throughfall and stemflow as well as input of S and N to the Ratanica forested catchment (S. Poland), which is exposed to moderate anthropogenic pollution are presented. There was high input of sulphur (26 kg ha−1) and nitrogen (24 kg ha−1) to the catchment, mainly in NH4+ (18 kg ha−1). The significant contribution of NH4 + connected with intensive agriculture in surrounding fields has led to eutrophication of the ecosystem.
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  • 26
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
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  • 27
    ISSN: 1432-0878
    Keywords: Pituitary ; Dexamethasone ; ACTH ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 3H-Dexamethasone (10 μg/kg) was injected intravenously in adrenalectomized rats and after survival times of 5, 30, 60, and 180 min its uptake within the pituitary was studied by autoradiography. Radioactivity was concentrated in cell nuclei in the pars nervosa and pars distalis. Within the pars intermedia, only cells of the marginal zone were labeled. In the pars distalis, some cells showed a weak nuclear accumulation of radioactivity as early as 5 min after injection. The tissue radioactivity was nearly maximal at 5 min, and the proportion of radioactivity in nuclei reached a maximum of 60–70% by 30 min. In competition experiments, non-radioactive steroids (1 mg/kg) were injected 5 min before 3H-dexamethasone and sacrifice was 30 min later. Dexamethasone markedly diminished the nuclear accumulation in the pars distalis, but corticosterone and progesterone did not. In the pars nervosa, corticosterone and progesterone competed for nuclear uptake of 3H-dexamethasone, although less effectively than dexamethasone itself. Different cell types in the pars distalis were characterized by treating autoradiograms with an immuno-peroxidase bridge procedure. Cells treated with anti-ACTH 17–39 had the greatest nuclear concentration of radioactivity, and those stained with anti-TSH were least heavily labeled. Cells treated with antisera to GH, PRL, and hCG were moderately labeled.
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  • 28
    ISSN: 1432-0878
    Keywords: Growth hormone-release inhibiting hormone ; Somatostatin ; Hypothalamus ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Utilizing the unlabeled antibody enzyme method, we investigated the distribution of hypothalamic elements immunoreactive with antibodies to growth hormonerelease inhibiting hormone (GH-RIH). Immunostained elements, resembling neural processes, are distributed along a pathway corresponding to a portion of the tuberoinfundibular tract. However, GH-RIH fibers are caudal, dorsal and medial to LH-RH fibers detected by the same technique. Similar topographic arrangements are noted in coronal and sagittal sections. Comparable results were obtained with two different preparations of antisera to GH-RIH. No cell bodies specifically stained by anti-GH-RIH were detected. Our data agree with those of other investigators using immunohistochemical techniques.
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  • 29
    ISSN: 1432-0878
    Keywords: Neurophysin ; Vasopressin ; Median eminence ; Electron microscopy ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With the use of tissue prepared by freeze-substitution and the unlabelled antibody enzyme technique, neurophysin and vasopressin were localized at the ultrastructural level in the posterior pituitary and median eminence of the guinea pig. In the posterior pituitary neurophysin was found in the large neurosecretory granules (1300–1500 Å) of axons, Herring bodies, and nerve terminals. In some of these axons immunoreactive neurophysin was found outside of granules in the axoplasm. By light microscopy neurophysin was found in both the zona interna and zona externa of the median eminence; this was confirmed by electron microscopy. In the zona interna as in the posterior pituitary, neurophysin was localized both inside and outside the large neurosecretory granules. In the zona externa, immunoreactive deposit was primarily located in granules with a diameter of 900–1100 Å in nerve terminals abutting on the primary portal plexus. The distribution of vasopressin paralleled that of neurophysin except that the hormone was rarely extragranular. These results demonstrate for the first time that both neurophysin and vasopressin are present in granules of axons that are in contact with the hypophysial portal vasculature.
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  • 30
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    Cell & tissue research 164 (1975), S. 543-557 
    ISSN: 1432-0878
    Keywords: Magnocellular hypothalamic nuclei ; Fetal and neonatal pigs ; Anti-porcine neurophysin serum ; Immunocytochemistry ; Slab gradient polyacrylamide gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The immunoperoxidase cytochemical reaction was applied to the localization of neurophysin-containing elements in the fetal and adult pig hypothalamus. In the 60 day fetal pig, cells of the supraoptic nucleus (SON) were the only structures in the hypothalamus in which neurophysin was detected. However, by 87 days the cell bodies in both the SON and paraventricular nucleus (PVN) contained neurophysin-like material. The distribution of immunoreactive material in the 111 day fetal animal was similar to that found in the adult pig. In transverse section of the mature pig the SON exists in two discrete components; an antero-lateral group of cells connected by scattered cells to a smaller postero-medial group. Anteriorly, the PVN appears as a line of cells bordering the third ventricle but as we proceed posteriorly the dorsal aspect expands laterally to give a wedge-shaped group of cells. In mid-sagittal sections, the cells of the PVN are distributed over a wide area of the anterior hypothalamus in a triangular profile. The borders between the SON and PVN became more difficult to define in medial sections than in lateral sections. Continuous gradient polyacrylamide gel electrophoresis was carried out on the neural lobe extracts from fetal, newborn and adult pigs. Proteins with an electrophoretic mobility similar to that of porcine neurophysins-I,-II and -III were present in the newborn and 98 day fetal pig. It is concluded that material immunoreactive with anti-neurophysin serum is present in the hypothalamus of the 60 day fetal pig. Furthermore, at late fetal development and during the postnatal period it is tentatively suggested that the neurophysin present in the pituitaries of these animals is chemically identical with that of adult neurophysin.
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  • 31
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    Cell & tissue research 178 (1977), S. 17-38 
    ISSN: 1432-0878
    Keywords: Myosin ; Immunocytochemistry ; Adrenal medulla ; Exocytosis ; Secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Myosin was isolated in high purity from the bovine adrenal medulla by gel filtration and ion exchange chromatography. The purified myosin was analyzed by electrophoresis in gels containing SDS and found to contain a 200,000 molecular weight heavy chain and major light chains of molecular weights 20,000 and 17,000 in a 1∶1∶1 molar ratio. At high ionic strength the myosin had high Ca-ATPase and K-EDTA-ATPase activities and low Mg-ATPase activity. At low ionic strength, the Mg-ATPase was activated to a low level by rabbit muscle actin. The myosin was found to decorate F-actin in the absence, but not the presence of ATP. In low ionic strength solutions, the myosin assembled into characteristic bipolar filaments. The distribution of this myosin in the adrenal medulla and of cross-reacting myosin in several other bovine tissues was determined with the use of antimedullary myosin immunoglobulin G as a specific stain that was detected by direct and indirect immunofluorescence. In the medulla strong staining was seen between the chords of chromaffin cells indicating the presence of a highly muscular vasculature that may perform functions analogous to those of the myoepithelium of exocrine glands. The chromaffin cells showed weak positive staining around the nuclei and in a pattern radiating toward adjacent blood vessels. Cells of the inner zone of the adrenal cortex showed strong staining in the peripheral cytoplasm while cells in the intermediate and outer zones did not stain. In a blood smear, platelets and the cytoplasm of leukocytes stained strongly while erythrocytes did not stain. In striated muscle and the gray and white matter of the cerebrum only the capillaries and larger vessels stained. In the liver the phagocytic cells bordering vascular sinuses stained strongly while the hepatocytes were separated from one another by a 2 micron trilaminar band possibly representing the microfilament web surrounding the bile canaliculi and associated with junctional complexes. The results suggest that myosin is present in several highly differentiated, non-motile tissue cells where it may play a role in secretion or other specialized functions.
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  • 32
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Cytology ; Pars tuberalis ; Immunocytochemistry ; Luteinizing hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The objective was to acquire evidence regarding the secretory capacity of cells in the pars tuberalis of the rat pituitary by the application of immunocytochemical staining. For this purpose the conjugated antibody and immunoglobulin-enzyme bridge techniques were utilized with antisera to the following hormones of the pars distalis: human somatotropin, human thyrotropin, human β-melanotropin, ovine luteinizing hormone (LH), porcine β17–39-corticotropin, and β1–24-corticotropin. Only LH-containing cells were demonstrated. They were exceedingly rare in the cephalic pars tuberalis beneath the median eminence. The frequency of LH-cells was greater in the pars tuberalis associated with the infundibulum and increased distally. LH-cells were most common ventrolateral to the infundibular stem and occurred singly and in clusters. These results indicate that following hypophysectomy the portion of the pars tuberalis that remains in situ has the capacity to secrete only LH of all the pars distalis hormones.
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  • 33
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    Cell & tissue research 186 (1978), S. 399-412 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Dog ; Pars distalis ; Thyrotropin (TSH) ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the immunoperoxidase technique and antisera to the specific beta (β) subunits of bovine and rat TSH1, selective immunocytochemical staining was localized in a specific cell population in the pars distalis of the dog pituitary gland. These TSH cells were found to be positive to aldehyde fuchsin, alcian blue, periodic acid-Schiff (PAS) and aniline blue. With the performic acidalcian blue (pH 0.2) -PAS-orange G procedure these cells stained blue-purple, demonstrating FSH/LH cells (blue or turquoise), ACTH/MSH cells (redpurple) and PRL cells (orange-red). The TSH cells were further differentiated from other functional cell types of the pars distalis on the basis of their typical cytological features, intraglandular distribution and by immunocytochemical double staining. In the pars distalis of adult male dogs the TSH cells were mostly shown to be smaller in size and less numerous than in bitches in the anestrous phase of the sexual cycle. Moreover, cytological alterations in the immunoreactive thyrotrophs in the pituitary of male and female dogs generally paralleled the spontaneous changes in thyroid function associated with thyroid atrophy and/or pituitary insufficiency, and thyroid hyperplasia or goiter. In conclusion, because of their specificity and high potency, the antisera to the β-subunits of bovine and rat TSH represent an effective tool for the selective immunocytochemical localization of TSH in the dog pituitary. This allows the study of the morphology and function of TSH cells under different physiological, pathological and experimental conditions.
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  • 34
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    Cell & tissue research 188 (1978), S. 99-106 
    ISSN: 1432-0878
    Keywords: LHRH-neurosecretion ; Avian hypothalamus ; Vasotocin neurosecretion ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A fluorescent technique applying specific LHRH and vasotocin antisera was used for the immunocytochemical localization of the respective neurosecretory systems in the hypothalamus of gonadectomized, testosteronetreated and/or serotonin injected male domestic ducks. An immunoreactive (IR) LHRH-producing system, with perikarya located in the preoptic nucleus, could be traced through the ventral hypothalamus down to the external layer of the rostral and caudal ME, in close vicinity to the hypophysial portal system. An IR-vasotocin system originating in the paraventricular and supraoptic nuclei ran through the ventral hypothalamus, but terminated in (i) the external layer of the rostral ME, and (ii) in the posterior lobe of the hypophysis.
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  • 35
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    Cell & tissue research 188 (1978), S. 119-132 
    ISSN: 1432-0878
    Keywords: Neurophysin ; Paraventricular nucleus ; Supraoptic nucleus ; Sheep ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum cross-reactive against ovine neurophysins-I-II and -III has been used in conjunction with the immunoperoxidase histochemical procedure to localize the cells of the sheep paraventricular (PVN) and supraoptic nuclei (SON). In order to describe the topographical distribution of the SON and PVN a study was made on the serial sections cut (a) transversely from rostral to caudal positions and (b) sagittally from lateral to medial positions of the hypothalamus. The cells of the SON, when examined in the transverse aspect, extended approximately 1900 μ caudally and when examined in the sagittal plane were contained within a lateral-medial distance of 4830 μ. In each case the SON cells lay adjacent to the optic chiasm. As sections were cut transversely, the cells of the PVN first appeared in a rostral position defined as 0 μ and close to the ventral lining of the third ventricle. This general ventral and ventro-lateral distribution of cells maintained up to a caudal distance of approximately 840 μ. From positions 1260–2310 μ there was a dramatic dorsal shift of the PVN cells which by this time had also extended laterally. The total rostral-caudal distance occupied by the PVN cells was 3150 μ. That the lateral-medial distance occupied by the PVN was small (1050 μ) was determined on examining the magnocellular nuclei in sagittal section.
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  • 36
    ISSN: 1432-0878
    Keywords: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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  • 37
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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  • 38
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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  • 39
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    Cell & tissue research 280 (1995), S. 541-548 
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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  • 40
    ISSN: 1432-0878
    Keywords: Key words: Melanin-concentrating hormone ; Immunocytochemistry ; Development ; ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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  • 41
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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  • 42
    ISSN: 1432-0878
    Keywords: Key words: Esophagus ; Epithelial cells ; Intestinal lectin ; L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells form a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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  • 43
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    Cell & tissue research 234 (1983), S. 427-437 
    ISSN: 1432-0878
    Keywords: Corticotropes ; Rat fetus ; Ultrastructure ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Corticotropes of rat fetuses aged 16, 18 and 21 days were localized by the indirect antibody-enzyme method on semithin sections of the pituitary. The development of the ultrastructure of these cells was observed on consecutive ultrathin sections. In comparison with previous data our present results show that identification of a fetal cell type cannot be based entirely on morphological criteria. The structural peculiarities of corticotropes obtained from studies in vivo are compared with those observed in cells maintained in vitro.
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  • 44
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    Cell & tissue research 234 (1983), S. 439-450 
    ISSN: 1432-0878
    Keywords: Avidin ; Avidin-biotin interaction ; Biotin affinity histochemistry ; Biotin hydrazide ; Immunocytochemistry ; Magnum gland ; Secretion ; Oviduct
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of endogenous avidin was studied cytochemically in the magnum tissue of the oviduct of laying hens. Two methods, based on an interaction of avidin-biotin with biotin hydrazide-peroxidase (B-HRP) as an affinity reagent, and on an immunoperoxidase technique, were tested by morphological analysis. The data obtained by both methods showed that in the magnum B-HRP is a strictly substitutive reagent for endogenous avidin. Avidin was clearly demonstrated in large amounts in the secretory granules of some epithelial cells and tubular gland cells, but was absent from mucous cells, the goblet cells, which had been believed to be the location of avidin production, and from ciliated cells. These granules had previously been demonstrated by both electron-microscopic cytochemical techniques. Especially in acinar cells, they were nonhomogeneous with a speckled core and a dense peripheral part. They ranged in size from 500 to 2200nm in diameter in the gland and 180 to 720 nm in the epithelium. Columnar epithelial cells containing avidin granules had a strong resemblance to those of the protodifferentiated tubular gland cells in the magnum of chicks pretreated with daily estrogen or estrogen plus progesterone, and might have migrated towards the acinus as substitutional secretory cells. Therefore, the acinar cells of the magnum, considered to be composed of several secretory protein-producing systems, are dependent on estrogen and/or progesterone in the oviduct of the laying hen.
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  • 45
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    Cell & tissue research 234 (1983), S. 547-559 
    ISSN: 1432-0878
    Keywords: Myelin proteolipids ; Oligodendrocytes ; Golgi apparatus ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Purified antibodies directed against myelin proteolipids were isolated by affinity chromatography of whole serum obtained from rabbits inoculated with myelin. These antibodies were specific for light, medium and dark oligodendrocytes. Astrocytes, neurons and their processes were not reactive. Immunocytochemical investigations showed that the membranes of the Golgi complex are highly labeled by these antibodies. Diffuse cytoplasmic labeling was only observed on the light and medium oligodendrocytes and was absent from the dark types. Vesicles possessing a punctate staining were detected in the vicinity of the Golgi complex and the oligodendroglial membrane. A discontinuous labeling of the plasmalemma appears to be characteristic of the actively myelinating light and medium oligodendrocytes. In compact myelin sheaths positive immunostaining was only detected at the dense line. The immunocytochemical localization of the myelin proteolipids in the oligodendrocytes is in accordance with previously published biochemical data.
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  • 46
    ISSN: 1432-0878
    Keywords: Key words: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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  • 47
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor cells ; Ion pumps ; Polarity ; Immunocytochemistry ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.
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  • 48
    ISSN: 1432-0878
    Keywords: Neurohemal areas ; Neuropeptides ; Monoamines ; Immunocytochemistry ; Nervous system, insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.
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  • 49
    ISSN: 1432-0878
    Keywords: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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  • 50
    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 51
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 52
    ISSN: 1432-0878
    Keywords: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their associated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in their encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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  • 53
    ISSN: 1432-0878
    Keywords: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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  • 54
    ISSN: 1432-0878
    Keywords: Esophagus ; Epithelial cells ; Intestinal lectin, L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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  • 55
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    Cell & tissue research 281 (1995), S. 101-108 
    ISSN: 1432-0878
    Keywords: Somite ; Intergin ; Extracellular matrix, structures ; Embryo ; Laminin ; Immunocytochemistry ; Macaca fascicularis (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of integrin subunits α6 and β1, and the α6β1 integrin ligand, laminin, was examined during somitogenesis in developmental stages 11, 13, and 16 in the long-tailed macaque, using peroxidase immunocytochemistry. Within differentiating somites in stage 11, α6 expression was observed in the sclerotome, basal surface of dermamyotomal cells adjacent to the basal lamina and on scattered cells throughout the dermamyotome. In further advanced somites in stages 13 and 16, α6 immunoreactivity become restricted to the myotome, α6 was expressed on mesenchymal core cells within the myocele of undifferentiated epitheliod somites and the ventromedial wall of somites commencing differentiation at each stage. β1 distribution resembled that of α6 in stage 11 somitic tissue, however, it remained present on myotome and sclerotome cells in the later stages, and was also expressed on dermatomal cells in stage 16. Laminin immunoreactivity, while more intense and prevalent than α6 and β1 in each stage examined, occurred on the same somite cell populations as the 2 integrin subunits. These results show a defined distribution of α6 on somitic tissue, and suggest this integrin is involved in somite differentiation. They also support a possible role for α6 in myoblast formation and migration. Overlapping of β1 and laminin immunoreactivity with that of α6 further suggests that α6 paris with β1 as a functional heterodimer for laminin in defined somitic regions.
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  • 56
    ISSN: 1432-0878
    Keywords: Key words: Gap junction ; Intercellular junction ; Insect ; Arthropod ; Immunocytochemistry ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the ∼10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an ∼40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an ∼30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.
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  • 57
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    Cell & tissue research 281 (1995), S. 507-515 
    ISSN: 1432-0878
    Keywords: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vitro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α-tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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  • 58
    ISSN: 1432-0878
    Keywords: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS), invertebrate ; Optic lobe ; Pigment-dispersing hormone, insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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  • 59
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development, ontogenetic ; Vitellointestinal duct ; Pancreas, exocrine ; Pancreas, endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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  • 60
    ISSN: 1432-0878
    Keywords: Key words: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS) ; invertebrate ; Optic lobe ; Pigment-dispersing hormone ; insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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  • 61
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    Cell & tissue research 233 (1983), S. 69-80 
    ISSN: 1432-0878
    Keywords: Somatostatin ; Hypothalamus ; Immunocytochemistry ; Human ; Rhesus monkey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical methods were used to compare the localization of somatostatin (SRIF) in the human and rhesus monkey hypothalamus. The distribution of SRIF-containing cell bodies and fibers is similar in the two species. Perikarya are located predominantly in the periventricular region and to a lesser extent in the ventromedial nucleus. Fibers occur in dense clusters within the periventricular region, ventromedial nucleus, arcuate nucleus, median eminence, and pericommissural area of both species. Analysis of serial sections suggests that fibers originate from cells in the periventricular region, extend ventrally through the ventromedial and arcuate nuclei to terminate around the portal vessels of the infundibular stalk, and thereby participate in the regulation of anterior pituitary function. Somatostatinergic fibers are also found surrounding non-immunoreactive perikarya in the ventromedial nucleus and periventricular region of both primates. This arrangement may support somatostatin's postulated role as a neurotransmitter or neuromodulator. The strong similarity between the localization of hypothalamic SRIF in the human and rhesus monkey supports the use of the rhesus monkey as a model for the study of somatostatin as a neuroendocrine regulator in the human.
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  • 62
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasopressin ; Dorsomedial hypothalamus ; Amygdala ; Locus coeruleus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Recently, the existence of a vasopressin-immunoreactive cell group was described in the bed nucleus of the stria terminalis (van Leeuwen and Caffé 1983). In the present investigation additional nuclei containing vasopressin-immunoreactive cells were found, after colchicine pretreatment, in the dorsomedial hypothalamus, medial amygdaloid nucleus and the locus coeruleus. Vasopressin-immunoreactive cells in the dorsomedial hypothalamus and medial amygdaloid nucleus are small (8–14 μm and 10–14 μm, respectively), while those in the locus coeruleus are medium-sized (20–25 μm). Incubation with anti-bovine neurophysin II and anti-rat neurophysin revealed staining of the same cell group in the above-mentioned areas. None of these cell groups show stained cells after incubation with anti-oxytocin and anti-bovine neurophysin I. When sections of the homozygous Brattleboro rat, which shows a deficiency in vasopressin synthesis, are incubated with anti-vasopressin, anti-bovine neurophysin II, or anti-rat neurophysin, no immunoreactivity can be observed in these brain regions. The above-mentioned cell groups may contribute to the vasopressinergic innervation of brain sites that have been reported to persist after lesioning of the suprachiasmatic, paraventricular and bed nuclei of the stria terminalis.
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  • 63
    ISSN: 1432-0878
    Keywords: α-Endorphin ; Immunocytochemistry ; Median neurosecretory cells ; Peptidergic neurones ; Calliphora vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A group of the 24–26 paraldehyde fuchsin-positive median neurosecretory cells (MNC) in the pars intercerebralis of the brain of the blowfly, Calliphora vomitoria, has shown immunoreactivity towards three different antibodies to α-endorphin, a peptide that corresponds to the amino acid sequence present between residues 61 and 76 of the precursor molecule, β-lipotropin (β-LPH). The immunoreactive material could be followed in axons within the median bundle, the tract through which neurosecretory material from the MNC is passed down to the corpus cardiacum (CC). The α-endorphin-immunoreactive material was observed leaving the CC in the cardiac-recurrent nerve, dorsal to the proventriculus, in the direction of the abdomen. The cells that contain the α-endorphin-like material are different from those of the MNC that contain insulin-, pancreatic polypeptide-, and gastrin/CCK-like peptides. This finding demonstrates the considerable complexity and peptidergic nature of the MNC and constitutes further evidence that morphinomimetic-like peptides are present in the nervous system of invertebrates.
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  • 64
    ISSN: 1432-0878
    Keywords: Angiotensinogen ; Liver ; Kidney ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The renin substrate, angiotensinogen, was localized by immunocytochemistry in liver and kidney of normal rats by the use of an antiserum directed against pure rat angiotensinogen. This substrate was also examined in rats after bilateral nephrectomy, which is known to increase plasma angiotensinogen, and in rats treated with colchicine, which inhibits serum protein secretion. In normal rat liver, light microscopy showed the presence of immunoreactive material in a very few cells. The number of stained hepatocytes rose in rats treated with colchicine or after bilateral nephrectomy. Immuno-staining increased further when rats were both nephrectomized and colchicine treated. In the kidney, angiotensinogen was specifically located as granular formations in nephrocytes of the proximal tubule but never in the granular cells of the juxtaglomerular apparatus. The localization of these granular formations under the brush border suggests that angiotensinogen is reabsorbed from the glomerular ultrafiltrate rather than synthesized in the kidney.
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  • 65
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    Cell & tissue research 233 (1983), S. 471-474 
    ISSN: 1432-0878
    Keywords: Prolactinergic neurons ; Immunocytochemistry ; Cerebral ganglion ; Styela plicata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peptidergic neurons have been detected in the cerebral ganglion of the ascidian Styela plicata by means of cytochemical methods. After incubation with a mammalian antibody to human prolactin the perikarya show a strong immunoreactivity. The possible function of prolactin-like peptides in the nervous system of protochordates is discussed.
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  • 66
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    Cell & tissue research 234 (1983), S. 125-134 
    ISSN: 1432-0878
    Keywords: Vasopressin ; Immunocytochemistry ; Electron microscopy ; Hypothalamus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vasopressin-containing neurons, identified by immunocytochemistry, are located predominantly in the posterior magnocellular division of the paraventricular nucleus of the rat hypothalamus. By electron microscopy, the immunoreaction product is seen within the cell bodies and neuronal processes. In the perikarya and dendritic processes, the immunoreactive material is associated primarily with neurosecretory granules. Axonal processes, identified by their content of microtubules and accumulation of neurosecretory granules, show the immunoreaction product in association with both of these organelles. Afferent axo-dendritic, axo-somatic and putative axo-axonic synapses with immunostained vasopressinergic neurons can be identified. The presynaptic profiles do not contain immunoreactive material. This study contributes to the ultrastructural characterization of vasopressinergic neurons in the paraventricular nucleus and of their afferent synaptic input.
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  • 67
    ISSN: 1432-0878
    Keywords: Fibronectin ; Lung ; Development ; Ultrastructure ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In a previous study changes in the macrodistribution of fibronectin during rat-lung development were examined. Using the peroxidase-antiperoxidase immunocytochemical technique, we have demonstrated the presence of fibronectin in embryonic, neonatal, and adult rat lung at the ultrastructural level. In the embryo, fibronectin is found both in an intra-and extracellular association with isolated pneumoblasts, and in a periodic distribution along the basal lamina. The neonate displays fibronectin in an intracellular association with early type-I cells and on their basal and luminal surfaces, but not in association with type-II cells. Neonatal basal lamina is diffusely labeled by anti-fibronectin antiserum. Fibronectin in adult tissue is found both intracellularly and on the basal and luminal surfaces of type-I cells but not in type-II cells. The basal lamina and interstitial connective tissue are slightly or non-reactive. These observations confirm and extend our initial suggestion that fibronectin is involved in rat-lung development.
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  • 68
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    Cell & tissue research 188 (1978), S. 259-264 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Human fetus ; Oxytocin ; Neurophysin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The use of antibodies against oxytocin or neurophysin enabled the detection by immunocytochemistry of oxytocin-neurophysin neurons in the hypothalamus in the human fetus. The perikarya of these neurons are located in the paraventricular and supraoptic nuclei. Immunoreactive neurons occur in the median eminence. The neurophysin immunoreactive neurons were more numerous than the oxytocin immunoreactive neurons. The specificity of the immunocytological reaction was controlled. The first oxytocin-neurophysin neurons are seen as early as the 14th week of gestation.
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  • 69
    ISSN: 1432-0878
    Keywords: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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  • 70
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    Cell & tissue research 280 (1995), S. 541-548 
    ISSN: 1432-0878
    Keywords: Musle, striated, skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 ν), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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  • 71
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    Cell & tissue research 280 (1995), S. 549-560 
    ISSN: 1432-0878
    Keywords: Enteric nervous system ; Immunocytochemistry ; Calretinin ; Calbindin ; Bombesin ; Small intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guineapig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin-and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.
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  • 72
    ISSN: 1432-0878
    Keywords: Laminin ; Nerve tracts ; Ontogenetic development ; Brain ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Laminin, a large glycoprotein of the basement membrane that promotes the growth of nerve cell processes in vitro has also been detected in the brains of developing embryos in situ where it is postulated to promote or guide neural outgrowth. We have investigated the histological and developmental patterns of a receptor to a specific pentapeptide sequence in the A chain of the laminin molecule (PA22-2 or IKVAV) that has been identified as a neuron growth-promoting sequence. Standard immunocytochemical procedures were used to localize the receptor by means of a polyclonal antibody to affinity-purified receptor (MR=110 kDa) from mouse brains. Results for postnatal stages (P) stages (P 1,7,8,25,30,and adult) show that the 110 kDa receptor is localized in fibers in the cortex and hippocampus, in astroglial cells at the surface of the cortex, and in neuronal cell bodies in the hippocampus. In contrast, the A-chain ligand is localized in cell bodies in the same regions at P stages. For embryonic stages (E) (E 14 and E 16) the receptor is localized in bundles of fibers in the superficial and deep cortical layers, and in cell bodies in these regions at E 14 only. Staining for the A chain ligand of the receptor was first seen postnatally. We speculate that the inverse histological pattern of receptor and ligand with respect to cell bodies and fibers may reflect a role in controlling axon guidance during development or repair during regeneration.
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  • 73
    ISSN: 1432-0878
    Keywords: Dopamine ; Serotonin ; Tyrosine hydroxylase ; Immunocytochemistry ; Nervous system, insect-Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine-(n〈-70) and serotonin-immunoreactive (n〈-120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine-and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.
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  • 74
    ISSN: 1432-0878
    Keywords: Cathepsin E ; Aspartic proteinase ; Osteoclasts ; Immunocytochemistry ; Rat (WKA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunocytochemical localization of cathepsin E, a non-lysosomal aspartic proteinase, was investigated in rat osteoclasts using the monospecific antibody to this protein. At the light-microscopic level, the preferential immunoreactivity for cathepsin E was found at high levels in active osteoclasts in the physiological bone modeling process. Neighboring osteoblastic cells were devoid of its immunoreactivity. At the electron-microscopic level, cathepsin E was exclusively confined to the apical plasma membrane at the ruffled border of active osteoclasts and the eroded bone surface. Cathepsin E was also concentrated in some endocytotic vacuoles of various sizes in the vicinity of the ruffled border membrane, some of which appeared to be secondary lysosomes containing the phagocytosed materials. These results strongly suggest that this enzyme is involved both in the extracellular degradation of the bone organic matrix and in the intracellular breakdown of the ingested substances in osteoclasts.
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  • 75
    ISSN: 1432-0878
    Keywords: Key words: Laminin ; Nerve tracts ; Ontogenetic development ; Brain ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Laminin, a large glycoprotein of the basement membrane that promotes the growth of nerve cell processes in vitro has also been detected in the brains of developing embryos in situ where it is postulated to promote or guide neural outgrowth. We have investigated the histological and developmental patterns of a receptor to a specific pentapeptide sequence in the A chain of the laminin molecule (PA22-2 or IKVAV) that has been identified as a neuron growth-promoting sequence. Standard immunocytochemical procedures were used to localize the receptor by means of a polyclonal antibody to affinity-purified receptor (MR=110 kDa) from mouse brains. Results for postnatal stages (P) stages (P 1,7,8,25,30,and adult) show that the 110 kDa receptor is localized in fibers in the cortex and hippocampus, in astroglial cells at the surface of the cortex, and in neuronal cell bodies in the hippocampus. In contrast, the A-chain ligand is localized in cell bodies in the same regions at P stages. For embryonic stages (E) (E 14 and E 16) the receptor is localized in bundles of fibers in the superficial and deep cortical layers, and in cell bodies in these regions at E 14 only. Staining for the A chain ligand of the receptor was first seen postnatally. We speculate that the inverse histological pattern of receptor and ligand with respect to cell bodies and fibers may reflect a role in controlling axon guidance during development or repair during regeneration.
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  • 76
    ISSN: 1432-0878
    Keywords: Key words: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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  • 77
    ISSN: 1432-0878
    Keywords: Key words: Choline acetyltransferase ; Cholinergic neuron ; Visual system ; Bolwig’s organ ; Immunocytochemistry ; In situ hybridization ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Choline acetyltransferase (ChAT) is the enzyme catalyzing the biosynthesis of acetylcholine and is considered to be a phenotypically specific marker for cholinergic neurons. We have examined the distribution of ChAT-expressing neurons in the larval nervous system of Drosophila melanogaster by three different but complementary techniques: in situ hybridization with a cRNA probe to ChAT messenger RNA, immunocytochemistry using a monoclonal anti-ChAT antibody, and X-gal staining of transformed animals carrying a reporter gene composed of 7.4  kb of 5′ flanking DNA from the ChAT gene fused to a lacZ reporter gene. All three techniques demonstrated ChAT-expressing neurons in the larval visual system. In embryos, the photoreceptor organ (Bolwig’s organ) exhibited strong cRNA hybridization signals. The optic lobe of late third-instar larvae displayed ChAT immunoreactivity in Bolwig’s nerve and a neuron close to the insertion site of the optic stalk. This neuron’s axon ran in parallel with Bolwig’s nerve to the larval optic neuropil. This neuron is likely to be a first-order interneuron of the larval visual system. Expression of the lacZ reporter gene was also detected in Bolwig’s organ and the neuron stained by anti-ChAT antibody. Our observations indicate that acetylcholine may be a neurotransmitter in the larval photoreceptor cells as well as in a first-order interneuron in the larval visual system of Drosophila melanogaster.
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  • 78
    ISSN: 1432-0878
    Keywords: Pituitary ; Gonadotrops ; Crinophagy ; Electron microscopy ; Enzyme cytochemistry ; Immunocytochemistry ; Autoradiography ; Catfish, Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The possible function of globules and irregular membrane-bound masses in the gonadotropin cells of the pituitary of Clarias gariepinus was studied. Strong secretory stimulation led to the disappearance of the secretory granules from gonadotropin cells but globules and irregular masses remained present. Acid phosphatase was detected enzyme-cytochemically in both globules and irregular masses. Radiolabelling with tritiated amino acids followed by autoradiography demonstrated that globules received radioactive material after secretory granules. The latter received radioactive material within 75 min of administration of radioactive amino acids but globules and irregular masses did not. Although some globules became radioactively labelled within 24 h of the administration of radioactive amino acids, irregular masses remained unlabelled during this period. Secretory granules reacted positively with antisera against α and β gonadotropin subunits, whereas globules and irregular masses only reacted with the antiserum against the β subunit. A moderate anti-7B2 immunoreactivity was demonstrated in secretory granules and globules, whereas irregular masses labelled strongly. The combined cytological results indicate that globules and irregular masses are degradative, possibly crinophagic structures which develop by fusional events from secretory granules to globules and then to irregular masses.
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  • 79
    ISSN: 1432-0878
    Keywords: Key words: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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  • 80
    ISSN: 1432-0878
    Keywords: Key words: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their ass ociated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in t heir encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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  • 81
    ISSN: 1432-0878
    Keywords: Key words: Pituitary ; Gonadotrops ; Crinophagy ; Electron microscopy ; Enzyme cytochemistry ; Immunocytochemistry ; Autoradiography ; Catfish ; Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The possible function of globules and irregular membrane-bound masses in the gonadotropin cells of the pituitary of Clarias gariepinus was studied. Strong secretory stimulation led to the disappearance of the secretory granules from gonadotropin cells but globules and irregular masses remained present. Acid phosphatase was detected enzyme-cytochemically in both globules and irregular masses. Radiolabelling with tritiated amino acids followed by autoradiography demons trated that globules received radioactive material after secretory granules. The latter received radioactive material within 75 min of administration of radioactive amino acids but globules and irregular masses did not. Although some globules became radioactively labelled within 24 h of the administration of radioactive amino acids, irregular masses remained unlabelled during this period. Secretory granules reacted positively with antisera against α and β gonadotropin subunits, whereas globules and irregular masses only reacted with the antiserum against the β subunit. A moderate anti-7B2 immunoreactivity was demonstrated in secretory granules and globules, whereas irregular masses labelled strongly. The combined cytological results indicate that globules and irregular masses are degradative, possibly crinophagic structures which develop by fusional events from secretory granules to globules and then to irregular masses.
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  • 82
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone ; Immunocytochemistry ; Development, ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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  • 83
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    Cell & tissue research 280 (1995), S. 123-131 
    ISSN: 1432-0878
    Keywords: Differentiation ; 5′-Nucleotidase ; Immunocytochemistry ; PC12 cells ; Synaptophysin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract 5′-Nucleotidase hydrolyzes 5′-mononucleotides to their nucleosides but is also thought to have a function in neuronal differentiation and synapse formation. The distribution of the enzyme, a glycosyl-phosphatidylinositol-anchored sialoglycoprotein, was investigated in PC12 cells using immunofluorescence microscopy. 5′-Nucleotidase was located both in intracellular compartments and at the cell surface. There was no principal difference in the cellular distribution between undifferentiated cells and after neuritogenic differentiation by nerve growth factor. Intracellularly, 5′-nucleotidase often revealed a sickle-shaped perinuclear distribution and a dotted pattern throughout the cytoplasm, including that of neurites and growth cones. The intracellular distribution was clearly different from that of the synaptic vesicle protein synaptophysin. However, the dotted fluorescence resembled that obtained after uptake of the endosomal marker acridine orange. 5′-Nucleotidase was present on the entire cell surface including all neurites formed after differentiation. There was no increase in 5′-nucleotidase fluorescence at synapse-like contacts between the tips of neurites and other PC12 cells. Surfacelocated 5′-nucleotidase could no longer be detected after the application of glycosyl-phosphatidylinositol-specific phospholipase C to cultured cells. This treatment did not affect PC12 cell differentiation. Our results thus reveal 5′-nucleotidase both at the surface and within organelles and suggest that PC12 cells may be used as a model system for the study of the physiological function of 5′-nucleotidase in neural cells.
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  • 84
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    Cell & tissue research 280 (1995), S. 123-131 
    ISSN: 1432-0878
    Keywords: Key words: Differentiation ; 5′-Nucleotidase ; Immunocytochemistry ; PC12 cells ; Synaptophysin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. 5′-Nucleotidase hydrolyzes 5′-mononucleotides to their nucleosides but is also thought to have a function in neuronal differentiation and synapse formation. The distribution of the enzyme, a glycosyl-phosphatidylinositol-anchored sialoglycoprotein, was investigated in PC12 cells using immunofluorescence microscopy. 5′-Nucleotidase was located both in intracellular compartments and at the cell surface. There was no principal difference in the cellular distrib ution between undifferentiated cells and after neuritogenic differentiation by nerve growth factor. Intracellularly, 5′-nucleotidase often revealed a sickle-shaped perinuclear distribution and a dotted pattern throughout the cytoplasm, including that of neurites and growth cones. The intracellular distribution was clearly different from that of the synaptic vesicle protein synaptophysin. However, the dotted fluorescence resembled that obtained after uptake of the endosomal marker acridine orange. 5′-Nucleotidase was present on the entire cell surface including all neurites formed after differentiation. There was no increase in 5′-nucleotidase fluorescence at synapse-like contacts between the tips of neurites and other PC12 cells. Surface-located 5′-nucleotidase could no longer be detected after the application of glycosyl-phosphatidylinositol-specific phospholipase C to cultured cells. This treatment did not affect PC12 cell differentiation. Our results thus reveal 5′ -nucleotidase both at the surface and within organelles and suggest that PC12 cells may be used as a model system for the study of the physiological function of 5′-nucleotidase in neural cells.
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  • 85
    ISSN: 1432-0878
    Keywords: Key words: Subcommissural organ ; Secretory glycoproteins ; Antibodies ; Immunochemistry ; Immunocytochemistry ; Dogfish ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The subcommissural organ of the dogfish, Scyliorhinus canicula (L), has been investigated by use of antibodies and lectins applied to blots and tissue sections processed for light and electron microscopy. Antibodies have been raised against each of the bands that have previously been identified in immunoblots by the use of antisera raised against secretory glycoproteins extracted from the dogfish subcommissural organ, viz., the 600-kDa band and two gel regions including the 475 to 400-kDa and the 145-kDa bands obtained from preparative gels; they are referred to as Ab-600, Ab-475/400, and Ab-145. These antisera and the lectins concanavalin A and wheat germ agglutinin have been used for the staining of: (1) blots of extracts of the dogfish subcommissural organ and optic tectum; (2) tissue sections of the dogfish brain. The findings indicate that the bands of 600, 475 and 400 kDa contain compounds that should be regarded as secretory glycoproteins of the dogfish subcommissural organ. The 600-kDa and 400-kDa bands are labeled by concanavalin A; wheat germ agglutinin labels the 475-kDa band strongly and the other two weakly. Ab-600 reacts with the bands at 600, 475 and 400 kDa and stains materials stored in the rough endoplasmic reticulum and secretory granules of 200–600 nm in diameter. The 600-kDa compound is probably a precursor form. Ab-475/400 stains the same three bands revealed by Ab-600; immunocytochemically, it reacts with two types of secretory granules (200–600 and 800–1200 nm in diameter) but it does not label the rough endoplasmic reticulum. Ab-145 reveals the bands at 600, 475 and 400 kDa and a diffuse zone in the region of 145 kDa; in light-microscopic immunocytochemistry, it behaves as Ab-475/400. The 475-kDa and 400-kDa glycoproteins, and a compound of approximately 145 kDa thus probably correspond to processed forms. Ab-475/400 stains granules present in cell processes ending on local blood vessels and at the leptomeninges. Since this antiserum selectively labels secretory granules, this finding may be taken as evidence for a basal route of secretion.
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  • 86
    ISSN: 1432-0878
    Keywords: Key words: Sauvagine ; Corticotropin-releasing factor ; Immunocytochemistry ; Interrenal gland ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunocytochemistry was used to investigate the presence of corticotropin-releasing factor-like peptides in the interrenal (adrenal) glands of the bullfrog Rana catesbeiana by using specific antisera raised against synthetic nonconjugated rat/human corticotropin-releasing factor, urotensin I, and sauvagine. From these three antisera, covering a broad range of corticotropin-releasing factor-like immunoreactivities, only the sauvagine antiserum gave positive immunoreactivity. Sauvagine immunoreactivity was found in cortical cells grouped into cords in the renal zone of the interrenal gland. The central and subcapsular cords were less stained. Tyrosine hydroxylase-positive chromaffin cells were not sauvagine-immunoreactive. The immunoreactivity was abolished, in all cases, by previous immunoabsorption of the sauvagine antiserum with synthetic sauvagine (0.1 μM), but it was not eliminated by sucker (Catostomus commersoni) urotensin I, sole (Hippoglossoides elassodon) urotensin I, sucker corticotropin-releasing factor, rat/human corticotropin-releasing factor, or ovine corticotropin-releasing factor (0.1–10 μM). In a sauvagine radioimmunoassay, interrenal extracts displaced 125I-sauvagine from antiserum only partially, and not in parallel with the sauvagine standard curve. The results suggest that the sauvagine immunoreactivity in the R. catesbeiana interrenal gland may represent a novel sauvagine-like peptide.
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  • 87
    ISSN: 1432-0878
    Keywords: Key words: Pituitary hormones ; Immunocytochemistry ; Morphometry ; Metamorphosis ; Bufo arenarum (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development and dynamics of thyrotropin (TSH), adrenocorticotropic hormone (ACTH), prolactin (PRL), and growth hormone (GH) cells have been studied using immunocytochemical techniques and rabbit antisera, raised against the relevant human hormone, in the pars distalis of Bufo arenarum larvae at different stages of development. The four types of cells studied were identified in different zones of the pars distalis: TSH cells occurred mainly in the centro-ventral zone, ACTH cells in the rostral and dorsal zones, GH cells in the central and caudal zones, and PRL cells in the anterior two-thirds of the gland. This distribution pattern does not show significant changes with development. Morphometry and stereology were used to evaluate the changes observed in the volume of the pars distalis and the immunoreactive cells during development. The former increased during larval growth and decreased throughout the metamorphic climax. The results obtained on cell number, volume density, and total volume suggest that, during larval growth (pre-prometamorphosis) of B. arenarum, TSH, PRL, GH and ACTH cells show a proliferative period with storage of their hormones; a second period involving hormone release occurs at the metamorphic climax.
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  • 88
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    Cell & tissue research 281 (1995), S. 367-374 
    ISSN: 1432-0878
    Keywords: Placenta ; Amniochorion ; Cytotrophoblast cells ; Immunoglobulin G ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Maternal immunoglobulin-G (IgG) is known to be transported across the placental syncytiotrophoblast during the period when the human fetus is incapable of manufacturing these defensive molecules. In this study we investigated the possible role of the amniochorion, that surrounds the amniotic cavity in which the fetus lies, in the transfer of immunoglobulin. Endogenous IgG was localised in the amniochorion by confocal immunofluorescence microscopy and by ultrastructural labelling of ultrathin frozen tissue sections using the protein A-gold technique. Immunoreactivity was identified in the extracellular matrix tissues and necrotic amniotic epithelial cells. Healthy amniotic epithelial cells and cytotrophoblast cells of the chorion laeve were devoid o endogenous IgG. These results suggest a possible non-specific paracellular transport pathway between cytotrophoblast cells, which may conceivably contribute to the acquisition of passive immunity by the fetus, and offer a rational explanation for the presence of small quantities of maternal IgG in the amniotic fluid.
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  • 89
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    Cell & tissue research 281 (1995), S. 507-515 
    ISSN: 1432-0878
    Keywords: Key words: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vi- tro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α−tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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  • 90
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    Cell & tissue research 185 (1977), S. 465-479 
    ISSN: 1432-0878
    Keywords: Somatostatin cells ; Pancreas ; Gut ; Immunocytochemistry ; Comparative study
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Somatostatin cells are numerous in the pancreas and digestive tract of mammals as well as birds. In the pancreas of chicken, cat and dog they occur in both the exocrine parenchyma and in the islets. In the rat and rabbit, somatostatin cells have a peripheral location in the islets, whereas in the cat, dog and man the cells are usually more randomly distributed. In the stomach of rabbits and pigs, somatostatin cells are more numerous in the oxyntic gland area than in the pyloric gland area, whereas the reverse is true for the cat, dog and man. In the cat, pig and man, somatostatin cells are fairly numerous in the duodenum, whereas in the rat, rabbit and dog they are few in this location. In the remainder of the intestines somatostatin cells are few but regularly observed. Somatostatin cells are numerous in the human fetal pancreas and gut. In the fetal rat, somatostatin cells first appear in the pancreas and duodenum (at about the 16–17th day of gestation) and subsequently in the remainder of the intestine. Somatostatin cells do not appear in the gastric mucosa until after birth. Three weeks after birth, somatostatin cells show the adult frequency of occurrence and pattern of distribution. In the chicken, somatostatin cells are numerous in the proventriculus, absent from the gizzard, abundant in the gizzard-duodenal junction (antrum), infrequent in the duodenum and virtually absent from the remainder of the intestines. No immunoreactive cells can be observed in the thyroid of any species nor in the ultimobranchial gland of the chicken. In the chick embryo, somatostatin cells are first detected in the pancreas and proventriculus (at about the 12th day of incubation). They appear in the remainder of the gut much later, in the duodenum at the 16th day, in the antrum at about the 19th day and still later in the lower small intestine. The ultrastructure of the somatostatin cells was studied in the chicken, rat, cat and man; the cells were identified by the consecutive semithin/ultrathin section technique. The somatostatin cells display the properties of the D cell. There was no difference in granule ultrastructure between somatostatin cells in the gut and the pancreas. The granules, which are the storage site of the peptide, are round, supplied with a tightly fitting membrane and have a moderately electron-dense, fine-granulated core. The mean diameter of the somatostatin granules is smallest in rat (155–170 nm) and largest in the chicken (270–290 nm).
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  • 91
    ISSN: 1432-0878
    Keywords: Key words: Nervous system ; insect ; DUM neuron ; Synapses ; Immunocytochemistry ; GABA ; Glutamate ; Locusta migratoria (Insecta) ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Dorsal unpaired median (DUM) neurones in the abdominal ganglia of the locust were impaled with microelectrodes and some were injected intracellularly with horseradish peroxidase so that their synapses could be identified in the electron microscope. Simultaneous recordings from DUM neurones in different abdominal ganglia revealed that they received common postsynaptic potentials from descending interneurones. Post-embedding immunocytochemistry using antibodies against GABA and glutamate was carried out on ganglia containing HRP-stained neurones. GABA-like immunoreactivity was found in 39% (n=82) of processes presynaptic to abdominal DUM neurones and glutamate-like immunoreactivity in 21% (n=42) of presynaptic processes. Output synapses from the DUM neurites were rarely observed within the neuropile. Structures resembling presynaptic dense bars but not associated with synaptic vesicles, were seen in some large diameter neurites.
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  • 92
    ISSN: 1432-0878
    Keywords: Key words: Leu-callatostatin ; Allatostatins ; Neuropeptides ; In situ hybridisation ; Immunocytochemistry ; Hindgut innervation ; Midgut endocrine cells ; Calliphora vomitoria ; Lucilia cuprina (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In situ hybridisation studies using a digoxigenin-labelled DNA probe encoding the Leu-callatostatin prohormone of the blowflies Calliphora vomitoria and Lucilia cuprina have revealed a variety of neurones in the brain and thoracico-abdominal ganglion, peripheral neurosecretory neurones, and endocrine cells of the midgut. With two exceptions, the hybridising cells are the same as those previously identified in immunocytochemical studies of sections and whole-mounts using Leu-callatostatin COOH-terminal-specific antisera. Within the brain and suboesophageal ganglion, there is a variety of neurones ranging from a single pair of large cells situated in the dorsal protocerebrum, to the several pairs of neurones in the tritocerebrum, some of which, in immunocytochemical preparations, can be seen to project via axons in the cervical connective to the thoracico-abdominal ganglion. In the medulla of the optic lobes, numerous small interneurones hybridise with the probe, as do clusters of similar-sized neurones close to the roots of the ocellar nerves. These results indicate that the Leu-callatostatin neuropeptides of the brain play a variety of roles in neurotransmission and neuromodulation. There are only three pairs of Leu-callatostatin-immunoreactive neurones in the thoracico-abdominal ganglion, at least two pairs of which project axons along the median abdominal nerve to provide extensive innervation of the hindgut. The Leu-callatostatin peripheral neurosecretory cells are located in close association with both nerve and muscle fibres in the thorax. In addition to neuronal Leu-callatostatin, the presence of the peptide and its mRNA has been demonstrated in endocrine cells in the posterior part of the midgut. These observations provide an example of a named brain/gut peptide in an insect.
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  • 93
    ISSN: 1432-0878
    Keywords: Key words: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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  • 94
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    Cell & tissue research 280 (1995), S. 549-560 
    ISSN: 1432-0878
    Keywords: Key words: Enteric nervous system ; Immunocytochemistry ; Calretinin ; Calbindin ; Bombesin ; Small intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guinea-pig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin- and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.
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  • 95
    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase ; Coronary vasculature ; Electron microscopy ; Immunocytochemistry ; Rat (Sprague Dawley) ; Guinea-pig (Dunkin Hartley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves closely associated with blood vessels in rat and guinea-pig hearts revealed many labelled nerve fibres in the walls of the main branches of the coronary arteries, and in arterioles, capillaries and post-capillary venules. The number of nitric oxide synthase-containing nerve fibres associated with different vessels, even those of the same calibre, varied. Terminal regions of nitric oxide synthase-immunoreactive fibres were observed in the endocardium and myocardium. Nitric oxide synthase-labelled fibres displayed electron-dense immunoproduct in both varicose and intervaricose regions. Immunoreactive axonal varicosities contained both small and large synaptic vesicles. The characteristics of the nitric oxide synthase-immunoreactive nerve fibres observed in the heart and the possibility that these fibres represent the processes of intracardiac neurones and/or sensory neurones of extrinsic origin are discussed.
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  • 96
    ISSN: 1432-0878
    Keywords: Key words: Dopamine ; Serotonin ; Tyrosine hydroxylase ; Immunocytochemistry ; Nervous system ; insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine- (n≤70) and serotonin-immunoreactive (n≤120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine- and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.
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  • 97
    ISSN: 1432-0878
    Keywords: Key words: Neurohemal areas ; Neuropeptides ; Monoamines ; Immunocytochemistry ; Nervous system ; insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin im-munoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.
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  • 98
    ISSN: 1432-0878
    Keywords: Key words: Compound eye ; Photoreceptor cells ; Ion pumps ; Polarity ; Immunocytochemistry ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.
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  • 99
    ISSN: 1432-0878
    Keywords: Key words: Blood platelets ; Immunocytochemistry ; Electron microscopy ; Coated vesicles ; Clathrin ; Adhesive proteins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Coated membranes and vesicles play an important role in receptor-mediated endocytosis and intracellular trafficking in various cell types, and are also present in blood platelets. Platelets take up certain proteins from the blood plasma, such as von Willebrand factor and fibrinogen, and these substances are transferred to storage granules. The receptors for these plasma proteins on the platelet plasma membrane have been well characterized, but morphological evidence for their transport to the storage granules is not yet available. In an attempt to clarify this aspect, we employed postembedding immunocytochemistry on platelets embedded in the acrylic resin LR White. Clathrin as the major coat component of coated vesicles was localized in the cytoplasm, on the plasmic faces of α-granules and the open canalicular system, and on the plasmic face of the plasma membrane. Colocalizations of the adhesive proteins, von Willebrand factor, fibrinogen and fibronectin, with clathrin could be observed at the same typical locations as coated vesicles were seen in Araldite-embedded material. These colocalizations have not been reported to date and furnish further evidence for a coated vesicle-mediated transport of blood plasma-derived adhesive proteins from their receptors on the outer plasma membrane to the α-granules.
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  • 100
    ISSN: 1432-0878
    Keywords: Blood platelets ; Immunocytochemistry ; Electron microscopy ; Coated vesicles ; Clathrin ; Adhesive proteins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Coated membranes and vesicles play an important role in receptor-mediated endocytosis and intracellular trafficking in various cell types, and are also present in blood platelets. Platelets take up certain proteins from the blood plasma, such as von Willebrand factor and fibrinogen, and these substances are transferred to storage granules. The receptors for these plasma proteins on the platelet plasma membrane have been well characterized, but morphological evidence for their transport to the storage granules is not yet available. In an attempt to clarify this aspect, we employed postembedding immunocytochemistry on platelets embedded in the acrylic resin LR White. Clathrin as the major coat component of coated vesicles was localized in the cytoplasm, on the plasmic faces of α-granules and the open canalicular system, and on the plasmic face of the plasma membrane. Colocalizations of the adhesive proteins, von Willebrand factor, fibrinogen and fibronectin, with clathrin could be observed at the same typical locations as coated vesicles were seen in Araldite-embedded material. These colocalizations have not been reported to date and furnish further evidence for a coated vesicle-mediated transport of blood plasma-derived adhesive proteins from their receptors on the outer plasma membrane to the α-granules.
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