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  • 1
    ISSN: 1432-0878
    Keywords: Parotid gland ; Membranes, Exocytosis ; Isoproterenol ; Amylase ; Stereology, Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A morphometric study has been made at the EM level of Isoproterenol (IPR) induced secretion of rabbit parotid glands in vivo. Emphasis has been placed here on the membrane content of acinar cells and the changes which occur following induced degranulation. In particular it was hoped to establish whether the preservation of zymogen granule membrane as intact electron microscopically visible subunits and the subsequent reutilisation of this membrane is a plausible hypothesis from a quantitative morphological standpoint. After two hours IPR had caused 〉95% depletion of granules. About 1343 μm2/cell of granule limiting membrane temporarily fused with the apical plasmalemma during this time and by two hours 1158 μm2/cell of this had been eliminated. Only a small increase in intracellular smooth membrane area was recorded after degranulation and we find no evidence that the zymogen granule membrane is stored indefinitely as smooth membrane fragments either in the region of the Golgi apparatus or elsewhere in the cytoplasm. IPR caused changes in RER membrane area (+37.7%, 1406 μm2/cell), which is a possible, but we consider implausible relocation site of granule membrane. The possible mechanism of the removal of ‘excess apical membrane’ and the ultimate fate of the zymogen granule membrane is discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Secretion granules ; Effect of feeding ; Carbachol ; Stereology ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The size, number and volume per cell of secretion granules in rat exocrine pancreas have been measured using stereological techniques. The changes which occur as a result of feeding starved animals (90 min) or stimulating lobular fragments in vitro with carbachol are documented. In fasted animals mean acinar cell volume was estimated as 1670 μm3 and the cells contained an average of around 450 secretion granules with a corrected mean diameter of 0.70 μm. They occupied around 7% of cell volume. After feeding mean cell volume was about 1300 μm3 and the cells contained an average of about 190 granules per cell with a mean diameter of 0.58 μm. They occupied 3% of cell volume. A shift in the size frequency distribution of granule diameters occurred as a result of feeding. In vitro experiments in which lobules were induced to secrete with carbachol (10μM, 3 h, 37° C) had a similar effect. Mean cell volume was reduced from around 1760 μm3 to 1360 μm3, mean granule number from around 420 per cell to 180 per cell and the volume density of granules was reduced from about 8% to 3% of cell volume. There was no significant change in mean granule diameter or shift in the size-frequency distribution of granule diameters. Incubation of tissues with cycloheximide (1 mM, 3 h, 37° C) did not prevent secretion by carbachol but it prevented replacement of granules. As a consequence, depletion by carbachol was greater in the presence of cycloheximide, the granules being reduced to around 110 per cell and to only 2.5% of cell volume. We conclude that feeding causes a preferential loss of larger granules and that during secretion replacement of granules occurs. Some of these granules are smaller than those evident in the glands of starved animals.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular histology 11 (1979), S. 19-50 
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Synopsis Centrifugation procedures have been evolved for isolating purified samples of rough endoplasmic reticulum, Golgi, zymogen granule and plasmalemmal membranes from homogenates of rabbit parotid gland tissue. The purification process was monitored using morphometry and enzyme and chemical marker assays. The membrane preparations were analysed by sodium dodecylsuphate (SDS) polyacrylamide gel electrophoresis, quantitative phospholipid thin layer chromatography and by enrichment studies. The results were used to evaluate various possible general models for the behaviour of membranes during the secretory cycle of parotid acinar cells.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rabbit polyclonal antibodies against isoproterenol-induced mouse proline-rich proteins (PRPs) were used to localize PRPs in the parotid salivary glands of normal adult BALB/c mice. The antibodies recognized both acidic-type and basic-type PRPs. Immunoblotting experiments revealed that the glands contained an acidic-type and a basic-type PRP. Parotid gland tissue was fixed with Karnosky's fixative and embedded in Lowicryl resin at low temperature. PRPs were localized at the electron microscope level using an indirect post-embedding staining technique with protein A-gold. The secretion granules of the acinar cells were strongly labelled. Pre-absorption of the antibody with purified acidic-type and basic-type PRPs indicated that the basic-type PRP is mainly located at the periphery of the granules but that the acidic-type PRP is more evenly distributed within the granules. Pre-absorption of the antibody with α-amylase did not affect the staining pattern, suggesting minimal cross-reactivity. PRPs were also detected within the rough endoplasmic reticulum and the Golgi apparatus of acinar cells, within the granules of the proacinar cells and in the lumena of the ducts, but not within the intercalated or striated duct cell granules.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Synopsis A quantitative study has been made on the enzymic, chemical and ultrastructural changes that occur in the parotid glands of rabbits as a result of Isoprenaline-induced secretion. Emphasis has been placed on correlating changes in organelle and membrane content which are evident 2 hr after Isoprenaline administration and which have been measured stereologically with the levels of appropriate enzymic or chemical markers, taking into account the contribution made by both the acinar and duct tissue. Lower protein, α-amylase and β-glycerophosphatase levels correlated with reductions in zymogen granule and lysosome volume whilst plasmalemmal and Golgi membrane areas and their marker enzyme concentrations remained unchanged. However, declines in alkaline phosphatase and succinate dehydrogenase activity (illustrated histochemically), andp-nitrophenyl phosphatase activity at pH 4.5 in the presence of tartrate occurred without any detectable decrease in membrane area. Conversely, an increase in rough endoplasmic reticulum area was measured stereologically but no increases in chemical markers were detected. The extent of correlation of the data is discussed in the context of the mechanism of secretion and the action of Isoprenaline.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Synopsis Rabbit parotid gland was chosen as a suitable model tissue for studying the role of membranes in enzyme secretion by acinar cells. The study was initiated using subcellular fractionation techniques. During these experiments the effects of various tissue disruption conditions such as the medium and the duration and vigour of homogenization were explored and the results assessed by enzyme and chemical assays and both quantitatively and qualitatively by electron microscopy. A series of basic fractions was isolated and marker enzyme or chemical assays selected for each of the relevant membrane types (rough endoplasmic reticulum, Golgi apparatus, zymogen granule, plasmalemma). A parallel study was effected using enzyme histochemical methods applied to frozen sections. Interesting comparisons could then be made between histochemical and biochemical methods of enzyme demonstration. These comparisons are discussed. the basic fractions provide the material from which specimens of purified membranes of the four species can be obtained. The isolation and characterization of such purified membranes is the subject of another report.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Synopsis A biochemical and histochemical study has been made of the characteristics and distribution of acid phosphatases in the parotid gland of the rabbit. The substratesp-nitrophenyl phosphate and β-glycerophosphate have been used. Homogenate studies showed that substantial hydrolysis ofp-nitrophenyl phosphate occurred in the presence of tartrate and fluoride ions but that β-glycerophosphate hydrolysis was almost abolished by these additives. The pH optima usingp-nitrophenyl phosphate and β-glycerophosphate were found to be around pH 5.0 and pH 4.0 respectively. Studies on subcellular fractions revealed that maximalp-nitrophenyl phosphate hydrolysis occurred in the microsomal fraction whereas maximal hydrolysis of β-glycerophosphate occurred in the mitochondrial fraction. The histochemical staining patterns produced by the two substrates differed in several respects. Whenp-nitrophenyl phosphate was used heavy plasmalemmal staining occurred, while reactions using β-glycerophosphate as substrate produced heavy staining of the acinar cell cytoplasm including many of the zymogen granules. Both substrates resulted in the staining of lysosomes and certain vesicles associated with the Golgi complex. Formaldehyde fixation resulted in an overall reduction in staining intensity and glutaraldehyde fixation used in electron microscopic studies heavily inhibited much of the extralysosomal activity. The conclusion is reached that significant extralysosomal acid hydrolase activity is present in the rabbit parotid gland, much of which is located on smooth microsomes. The data is discussed in the context of the distribution of extralysosomal acid hydrolases and the staining patterns are considered in the light of the limitations inherent in the techniques employed.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 209 (1980), S. 315-327 
    ISSN: 1432-0878
    Keywords: Rabbit ; Parotid ; Stereology ; Isoprenaline ; Secretion granules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A detailed stereological analysis has been made of the organelle content of rabbit acinar cells during the restoration of granule stores following extensive degranulation with isoprenaline (IPR). Rabbits were sacrificed 2, 4, 8, 12 and 16 h after IPR administration and the volumes and proportions of intracellular organelles were compared with those of untreated glands. At 2 h only 5% of cell volume was occupied by secretion granules, but there was already evidence of nascent granule formation. The volume per cell of secretion granules increased in sigmoid fashion and by 16 h amounted to 350 μm3/cell, 36% of cell volume, which are values similar to those of the control replete glands. IPR treatment caused some initial swelling of the cells, and there were transient increases in the volumes of several compartments. However, the volume of mitochondrial and lysosomal compartments had returned to control levels by 4 h and that of the nucleus by 8 h. The greatest increase was in the volume of the rough endoplasmic reticulum which had increased by nearly 70% by 2 h and remained enlarged throughout the period of restitution. However, neither the volume nor the proportion of the smooth membraned compartment varied throughout the period of analysis. The results are analysed in the light of the overall response of the cells to IPR and the interaction of the organelles during the synthetic phase of the secretory cycle. They are presented as a basis for ensuing studies of the granule populations and the membrane composition of the cells during the restoration of granule stores.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 199 (1981), S. 389-401 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Recently weaned male rabbits were injected either with 150 μg/kg isoprenaline in saline containing 0.01 M ascorbic acid or simply with the drug vehicle. Groups of drug-injected animals were killed at various time after injection. Parotid gland tissue samples from all animals were fixed, embedded and thin sectioned, and micrographs were prepared at standard magnification. Estimations of membrane areas of each membrane type in parotid acinar cells were made. It was found that in animals killed 2 hours after induced secretion apical area was larger than in controls. In animals killed at sucessively later times the apical area was progressively less. No elevation of any internal smooth membrane areas was ascertained at any sampling time, though the areas of rough endoplasmic reticulum in 2-12 hour samples were larger. It is suggested that excess apical membrane, though probably removed by interiorization, is afterwards disassembled inside the cell to create fresh macromolecular building units (protein molecules), perhaps after passing through the Golgi apparatus. This cryptic pool of building units can provide about 900 μm2 of secretion granule membrane per cell, the supply apparently being exhausted in the first eight hours after degranulation, whilst granule numbers are being increased. Thereafter, apparently, limited granule fusion occurs, so that ultimately the cellular complement of secretion granule membrane comes to enclose a greater volume of secretory product, though the average granule number per cell is smaller.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 199 (1981), S. 377-387 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The development of the secretion granule population of rabbit parotid glands after isoprenaline (IPR)-induced degranulation has been analyzed at the E.M. level using stereological techniques. Young male New Zealand White rabbits were sacrificed 2, 4, 8, 12 and 16 hours after IPR administration and the granule populations compared with those of starved controls. In controls. In control glands a third of cell volume was occupied by stored secretory material, and it was estimated that, on average, cells contained 386 granules. The granule population as a whole had a mean diameter of 0.94 μm, with a unimodal positively skewed size distribution. Two hours after IPR treatment overall granule volume density was only 15% of that of control glands, but there was evidence that the process of restitution had already begun. At 8 hours about a fifth of acinar cell volume was occupied by electron-dense granules with an estimated mean diameter of only 0.58 μm, and the population as a whole was more strongly skewed than in the controls. In the later stages of restitution (12 and 16 hours), the volume of stored secretory material continued to rise, mean granule diameter increased, the size-frequency distribution became less skewed and the estimated number of granules per cell fell to 277 by 16 hours, suggesting that some granule fusion occurs during development.The analyses are discussed in relation to the techniques employed, and the results are equated with other independent evidence of the mode of granule genesis.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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