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1.
Localisation of -amylase (EC 3.2.1.1) in barley aleurone cells treated with gibberellic acid has been achieved using protein A-gold-labelled polyclonal antibodies. Gold particles were located almost exclusively over the lumen of the rough endoplasmic reticulum and cisternae of the Golgi apparatus. The label was most concentrated over the Golgi apparatus. This indicates that the Golgi is involved in the secretion of -amylase protein from aleurone cells.Abbreviations ER endoplasmic reticulum - GA3 gibberellic acid - PBS phosphate-buffered saline  相似文献   

2.
H. Lehmann  D. Schulz 《Planta》1969,85(4):313-325
Summary In meristematic cells of the gemma of Riella helicophylla and in young bud cells from the protonema of Funaria hygrometrica the cell plate is formed by fusion of small vesicles originating from the Golgi apparatus. These spherical vesicles of about 0.1 m diameter have an electron dense centre, probably consisting of pectic substances or their precursors. The endoplasmic reticulum producing multivesicular bodies participate in cell plate formation too. Another cytoplasmic component forming the cell plate are coated vesicles, the origin of which is the Golgi apparatus and perhaps also the endoplasmic reticulum. In view of these observations the question of whether the endoplasmic reticulum or the Golgi apparatus forms the cell plate must be answered in this way: both endoplasmic reticulum and Golgi apparatus supply material for growth of the cell plate. Multivesicular bodies, coated vesicles and other small vesicles of unknown nature participate in the formation of the primary wall.

Zum Teil finanziert mit Sondermitteln des Landes Niedersachsen an Prof. Dr. M. Bopp.  相似文献   

3.
Summary The cytochemical localization of acid phosphatase and thiamine pyrophosphatase activity was studied by light and electron microscopy in prothoracic gland cells of the cockroach Leucophaea moderae. Nymphal and young adult animals were used.Prominent sites of acid phosphatase activity included large membrane-bounded dense bodies or lysosomes, and certain cisternae of the Golgi apparatus. The results suggest a possible difference in the enzymatic activity toward glycerophosphate and aromatic phosphates as substrates.Thiamine pyrophosphatase activity was localized in elements of the Golgi apparatus and endoplasmic reticulum, and in lysosome-like dense bodies. This latter activity was abolished by sodium fluoride treatment, whereas the phosphatase activity in the Golgi apparatus and endoplasmic reticulum is unaffected by such inhibition.The cytochemical results confirm through direct evidence the suggestions of Scharrer (1964), that the large dense bodies present in the prothoracic gland cells are lysosomes, and that their activity may be related to stages in the life history of the glands. Furthermore, the lysosomes or their derivative structures may play an essential role in the autolysis of the prothoracic glands toward the end of their active period.The enzymatic activity of the endoplasmic reticulum may indicate the involvement of this organelle in the metabolism of steroid-like precursor materials necessary for the synthesis of ecdysone.This study was supported by U.S.P.H.S. grants 5 T1-MH-6418 and NB-05219, and grant RO 1-AM-3984 to Dr. Berta Scharrer. I would like to express my appreciation to Dr. Scharrer for her encouragement and assistance during this study. I also wish to thank Mrs. Sarah Wurzelmann for her competent technical aid.  相似文献   

4.
Summary Immunoreaction of -fetoprotein (AFP) was detected not only in well-differentiated hepatocellular carcinoma but also in hepatocytes forming foci in livers with hyperplastic nodules during 3-methyl-4-dimethylaminoazobenzene hepatocarcinogenesis. The subcellular location of AFP in hepatoma cells was in the rough endoplasmic reticulum, perinuclear space and well-developed Golgi apparatus around the nucleus. In livers with hyperplastic nodules it was also in some parts of the smooth endoplasmic reticulum and Golgi regions in hepatocytes in the vicinity of submembranous areas or bile canaliculi. These findings suggest that the Golgi apparatus in hepatoma cells acts mainly as an organelle for glycosylation of AFP and that the Golgi complexes in the hepatocytes in livers with hyperplastic nodules are organelles for secretion of AFP.Combined light microscopic immunoperoxidase study and autoradiography with 3H-thymidine revealed a higher cumulative labeling index in AFP-positive hepatoma cells than in non-tumorous areas. Combined electron microscopic immunoperoxidase study and autoradiography showed that hepatoma cells with AFP immunoreactivity only in the rough endoplasmic reticulum had a significantly higher labeling index than did cells with AFP immunoreactivity in both rough endoplasmic reticulum and Golgi apparatus. These findings suggest that AFP is synthesized in hepatoma cells before or during the stage of their DNA synthesis and is then transported to the Golgi apparatus.  相似文献   

5.
D. N. Young 《Protoplasma》1978,94(1-2):109-126
Summary Clusters of small, pyriform gland or vesiculate cells protrude from the inner cortex into the inflated bladders ofBotryocladia pseudodichotoma. These multinucleate cells contains a large, central vacuole surrounded by dense cytoplasm. The lobed chloroplasts lack phycobilisomes and contain concentric rather than parallel thylakoids. Irregularly shaped, osmiophilic bodies of undetermined composition are abundant in the cytoplasm and are also released into the vacuole. The cell is characterized by abundant Golgi apparatus. The Golgi bodies appear to form by coalescence of endoplasmic reticulum derived vesicles and tubules. The Golgi apparatus produces large, electron transparent vesicles that contain an osmiophilic, fibrous material. This material is secreted into a diffuse wall layer which appears to be continually sloughed. Thus, the vesiculate cells ofBotryocladia possess an ultrastructure characteristic of granulocrine secretory cells, and it is suggested that they produce the slime contained within the bladders of this genus.  相似文献   

6.
The protein kinase CK2 is composed of two catalytic - or - and two regulatory -subunits. In mammalian cells there is ample evidence for the presence of individual CK2 subunits beside the holoenzyme. By immunofluorescence studies using peptide antibodies which allow us to detect the CK2-, - and -subunits we found all three subunits to be co-localized with a 58 KDa Golgi protein which is specific for the Golgi complex. Subfractionation studies using dog pancreas cells revealed the presence of all three subunits of CK2 at the smooth endoplasmic reticulum (sER)/Golgi fraction whereas the rough endoplasmic reticulum (rER) harboured only the catalytic - and -subunits. We found that the microsomal preparation from dog pancreas cells contained CK2 which phosphorylated a CK2 specific synthetic peptide and which was heparin sensitive. Furthermore, we could immunoprecipitate the CK2-subunit that exhibited a kinase activity which phosphorylated a CK2 specific substrate and which was heparin sensitive. Protease digestion experiments revealed that the CK2 subunits were located on the cytosolic side of the rER and the sER/Golgi complex. Thus, we could demonstrate an asymmetric distribution of the CK2 subunits at the rER and sER/Golgi complex. Since the CK2- and -subunits exhibit a substrate specificity which is different from the CK2 holoenzyme one might speculate that the asymmetric distribution of the CK2 holoenzyme and the CK2 catalytic subunits may have regulatory functions.  相似文献   

7.
Summary The acid hydrolases -glucosidase, -galactosidase,N-acetyl--d-hexosaminidase, -glucocerebrosidase and cathepsin D were studied immunocytochemically in normal and mutant human cells using monoclonal and affinity-purified polyclonal antibodies. For light microscopy, Rhodamine or Fluorescein-labelled conjugates were used, and for electron microscopy protein A-gold conjugates were employed. With the double labelling procedure, it was found that in normal fibroblasts every lysosome contained all the enzymes studied. The method described also enabled us to demonstrate the presence or absence of mutant enzyme protein in fibroblasts derived from patients with a genetic lysosomal enzyme deficiency.Immunoreactive acid hydrolases or their precursor forms were found in the rough endoplasmic reticulum, the cisternae of the Golgi complex, Golgi associated vesicles and lysosomes. This is in agreement with the present concept that the Golgi complex plays an essential role in the processing and targeting of lysosomal enzymes.  相似文献   

8.
Summary Ovaries were obtained from normal adult dairy cows at all days of the estrous cycle. The largest Graafian follicle and corpus luteum were excised and prepared for electron microscopic study.In the follicle wall, membrana granulosa cells contained granular endoplasmic reticulum and mitochondria with villous or lamellar cristae. The theca interna cells during proestrus and estrus contained ribosomes separated from endoplasmic reticulum. The latter during these periods assumed tubular and tortuous shapes. Mitochondria during these periods assumed rounded shapes, were occasionally cup-shaped, and developed tubular cristae.In the corpus luteum, the large luteal cells during metestrus and diestrus contained an abundance of agranular, tubular, branching membranes of endoplasmic reticulum and Golgi apparatus. Mitochondria were large, with tubular cristae, but smaller mitochondria, with irregular or villous cristae, were also present. Transitional bodies of the latter mitochondria to another form were observed. Cup-shaped and annular mitochondria were present during diestrus. In the small luteal cells, large vesicular membrane formations were present and often associated with lipid bodies. The cells were lipid-laden. Lysosomes and granular bodies were present during luteal regression.The observed features of the granulosa cells are related to protein synthesis, those of the pre-ovulatory theca interna cells and metestrus-diestrus large luteal cells to steroid synthesis, and those of the small luteal cells to lipid storage.This investigation was supported by a General Research Support Grant to the College of Veterinary Medicine, University of Minnesota, and Research Grant No. GM-07009, of the United States Public Health Service. Approved for publication as Scientific Journal Series Paper No. 6344, Minnesota Agricultural Experiment Station. The work reported is taken from the senior author's Ph. D. thesis.  相似文献   

9.
Summary The localization of -amylase (EC 3.2.1.1) in barley (Hordeum vulgare L. cv Himalaya) aleurone protoplasts was studied using electron microscope immunocytochemistry. Antibodies were raised against total barley -amylase, i.e., -amylase containing both highisoelectric point (high-pI) and low-pI isoforms, as well as against purified high- and low-pI isoforms. All antibodies localized -amylase to the endoplasmic reticulum (ER) and Golgi apparatus (GApp) of the aleurone cell, and various controls showed that the labeling was specific for -amylase. Labeling of protein bodies and spherosomes, which are the most abundant organelles in this cell, was very low. There was no evidence that -amylase isoforms were differentially distributed within different compartments of the endomembrane system. Rather, both high- and low-pI isoforms showed the same pattern of distribution in ER and in the cis, medial, and transregions of the GApp. We conclude that in the Himalaya cultivar of barley, all isoforms of -amylase are transported to the plasma membrane via the GApp.Abbreviations ER endoplasmic reticulum - GA3 gibberellic acid - GApp Golgi apparatus - PBS phosphate buffered saline - PCR partially coated reticulum - PM plasma membrane - TBS Tris buffered saline - TGN trans-Golgi network  相似文献   

10.
Summary Microscopy techniques were used to identify the pathway of transport of soybean leaf vegetative storage proteins (VSP/ and VSP94) to the vacuoles of a specialized cell type, the paraveinal mesophyll (PVM), where they accumulate. PVM cells are enriched in endoplasmic reticulum and Golgi bodies relative to surrounding mesophyll cells. The margins of medial and trans Golgi cisternae had attached or closely associated noncoated vesicles with densely staining membranes and lumenal contents of the same appearance as material that accumulated in the vacuole. These vesicles appeared to be transported preferentially to the tonoplast, where fusion with the membrane released the granular contents into the vacuole. Cytochemical staining with phosphotungstic acid and silver methenamine supported this interpretation as both the Golgi vesicles and the tonoplast stained intensely with these reagents, unlike the tonoplast of mesophyll cells which do not accumulate VSP. Immunocytochemical localization for VSP/ labeled the Golgi bodies and associated vesicles, and vacuolar material in PVM cells, but not in mesophyll. Similar labeling was seen in PVM of another legume species previously found to accumulate antigenically similar VSPs. Immunolocalization for VSP94, a lipoxygenase, labeled the PVM cytosol and material in the PVM vacuole, but not the Golgi or vesicles. The results of this study demonstrate that the Golgi pathway is utilized for transport of VSP/ in the PVM, which follows the mechanism of deposition demonstrated for certain seed storage proteins. VSP94 appeared to follow a separate path for accumulation in PVM vacuoles.Abbreviations LOX lipoxygenase - PVM paraveinal mesophyll - RER rough endoplasmic reticulum - TEM transmission electron  相似文献   

11.
Summary Arabinogalactan proteins (AGPs) are proteoglycans detected in high amounts at plant cell surfaces; however, details of their subcellular localization are largely unknown. Immunolocalization studies with the anti-AGP monoclonal antibody LM2 have indicated that this AGP epitope is associated with secretory compartments such as endoplasmic reticulum and Golgi apparatus within plant cells actively producing and secreting AGPs. The LM2 epitope contains a -linked glucuronic acid residue and occurs in the polysaccharide moiety of AGPs. We have localized this AGP epitope also to the tonoplast and to cytoplasmic strands. Endomembrane association of AGPs was confirmed with two other monoclonal antibodies, JIM13 and MAC207, both reacting with carbohydrate AGP epitopes containing GlcpA-(13)-D-GalpA-(12)-L-Rha residues. Immunocytochemistry is supported by biochemical analysis which shows that LM2 reacts with the microsomal fraction and also with low-molecular-weight material of the detergent phase after Triton X-114 phase separation prepared from maize roots. Our results indicate that some AGP epitopes are closely associated with endomembranes.Abbreviations AGP arabinogalactan protein - ER endoplasmic reticulum - GlcA glucuronic acid Dedicated to Professor Walter Gustav Url on the occasion of his 70th birthday  相似文献   

12.
Summary The localization of 3-hydroxysteroid dehydrogenase/isomerase (3-HSD) was studied in bovine adrenal glands by light as well as electron microscopic immunocytochemistry, using anti-bovine adrenal 3-HSD antibody. With light microscopy the cytoplasm of the glomerulosa cells was weakly immunostained, while that of the fasciculata-reticularis cells was intensely immunostained though both the capsular connective tissue cells and the medullary cells were entirely negative for this reaction. Electron microscopic immunocytochemistry revealed that the positive reaction products for 3-HSD were present on the membrane of smooth endoplasmic reticulum of the cortical cells, especially that of the fasciculata and reticularis cells. Other cell organelles such as mitochondria and Golgi apparatus were entirely negative. The present results indicate that 3-HSD is present in the membrane of smooth endoplasmic reticulum of bovine adrenal cortical cells.Supported by grants from the Ministry of Education Science and Culture, Japan  相似文献   

13.
Summary The localization of inhibin -subunit in the human testis was studied at the light- and electron-microscope level with immunostaining techniques. Antibodies against specific fragments of porcine and human inhibin -subunits were utilized. At light microscopy, inhibin -subunit immunoreactivity was detected in Sertoli cells, spermatocytes and in some Leydig cells. At electron microscopy, gold labeling was found in the cisternae of the Golgi apparatus and in the endoplasmic reticulum of Sertoli and Leydig cells. Gold labeling for inhibin was also found in coated vesicles in the cytoplasm of Sertoli cells as well as in coated pits and coated vesicles in the cytoplasm of some spermatocytes. The results of the present study suggest that, in the human testis, inhibin is produced by Sertoli and Leydig cells and is taken up by spermatocytes, on which it might act in a paracrine manner.  相似文献   

14.
We have studied by high resolutionin situ light and electron microscopic lectin-gold techniques the subcellular distribution of -d-Gal residues using theGriffonia simplicifolia I-B4 isolectin and compared it with that of -d-Gal residues as detected with theDatura stramonium lectin in Ehrlich tumour cells grown as ascites or monolayer. The microvillar but not the smooth plasma membrane regions were labelled with theGriffonia simplicifolia I-B4 isolectin whereas both plasma membrane regions were equally well labelled with theDatura stramonium lectin. Elements of the endocytotic/lysosomal system such as coated membrane invaginations and vesicles, early and late endosomes and secondary lysosomes were positive for both -d-Gal and -d-Gal residues. A particular feature of Ehrlich tumour cells is an elaborate tubular membrane system located in the pericentriolar region which is labelled throughout by both lectins and represents part of the endosomal system. In the Golgi apparatus labelling with both lectins was observed to commence in trans cisternae which is indirect evidence for a joint distribution of the sequentially acting 1,4 and 1,3-galactosyl-transferases.  相似文献   

15.
Dry, fresh uredospores ofPuccinia recondita that have been killed by infra-red radiation showed no striking ultrastructural differences from dry, fresh, viable uredospores. Numerous spherical and elipsoidal mitochondria with distinct and deep cristae, ribosomes, endoplasmic reticulum and convoluted plasmalemma were well shown by both.When moistened and incubated, killed uredospores lost ultrastructural organization, whereas moistened, viable, fresh uredospores imbibed moisture, became more spherical and germination commenced. A more or less centrally located nucleus was seen with the double membrane invaginated at some points. The advance of the germ tube was initially by enzymic degradation and concluded by mechanical disruption of the degraded pore plug. The mitochondria and the endoplasmic reticulum increased in number and the stored lipid bodies were gradually depleted as germination progressed.Features known as the foamy cytoplasm and folded membranes were seen in the germinating uredospores only. It was suggested that the foamy cytoplasm could be functionally similar to the glyoxisome because of the close association of the former to lipid bodies. The folded membranes may be accumulated endoplasmic reticulum being transported to the site of wall formation.Abbreviations used in electronmicrographs CW Cell wall - er Endoplasmic reticulum - f folded membranes - GP Germ pore - L Lipid bodies - M Mitochondria - P Plasmalemma - Pm foamy cytoplasm - R Ribosomes - T Germ tube  相似文献   

16.
Summary The morphology of the absorbing cells of the duodenal villi in the mouse, the rat, the hamster and the guinea-pig is described. The polymorphism of the dense bodies is pointed out. The fine localization of acid and alkaline phosphatase is investigated and compared. In all the species, acid phosphatase activity is observed in the dense bodies, Golgi vesicles and rare smooth endoplasmic profiles. Alkaline phosphatase is localized on the microvilli, Golgi apparatus, some smooth endoplasmic cisternae and numerous dense bodies. The presence of an alkaline phosphatase reaction in the dense bodies, probably lysosomes, of the absorbing cells is discussed. It is assumed that this enzyme follows a catabolic pathway and is finally degraded in the lysosomes.Abbreviations used AlPase alkaline phosphatase - AcPase acid phosphatase This work was done thanks to the contract C.E.N./A.I.E.A. N 347/RB and thanks to grants from the Fonds de la Recherche scientifique fondamentale collective.  相似文献   

17.
Summary We examined the intracellular localization of sugar residues of the rat gastric surface mucous cells in relation to the functional polarity of the cell organellae using preembedding method with several lectins.In the surface mucous cells, the nuclear envelope and rough endoplasmic reticulum (rER) and cis cisternae of the Golgi stacks were intensely stained with Maclura pomifera (MPA), which is specific to -Gal and GalNAc residues. In the Golgi apparatus, one or two cis side cisternae were stained with MPA and Dolichos biflorus (DBA) which is specific to terminal -N-acetylgalactosamine residues, while the intermediate lamellae were intensely labeled with Arachis hypogaea (PNA) which is specific to Gal 1,3 GalNAc. Cisternae of the trans Golgi region were also stained with MPA, Ricinus communis I (RCA I) which is specific to -Gal and Limax flavus (LFA) which is specific to -NeuAc. Immature mucous granules which are contiguous with the trans Golgi lamellae were weakly stained with RCA I, while LFA stained both immature and mature granules.The differences between each lectin's reactivity in the rough endoplasmic reticulum, in each compartment of the Golgi lamellae and in the secretory granules suggest that there are compositional and structural differences between the glycoconjugates in the respective cell organellae, reflecting the various processes of glycosylation in the gastric surface mucous cells.  相似文献   

18.
Walker LM  Sack FD 《Protoplasma》1995,189(3-4):238-248
Summary A distinctive feature of tip-growing plant cells is that cell components are distributed differentially along the length of the cell, although most ultrastructural analyses have been qualitative. The longitudinal distribution of cell components was studied both qualitatively and quantitatively in the apical cell of dark-grown protonemata of the mossCeratodon. The first 35 m of the apical cell was analyzed stereologically using transmission electron microscopy. There were four types of distributions along the cell's axis, three of them differential: (1) tubular endoplasmic reticulum was evenly distributed, (2) cisternal endoplasmic reticulum and Golgi vesicles were distributed in a tip-to-base gradient, (3) plastids, vacuoles, and Golgi stacks were enriched in specific areas, although the locations of the enrichments varied, and (4) mitochondria were excluded in the tipmost 5 m and evenly distributed throughout the remaining 30 m. This study provides one of the most comprehensive quantitative, ultrastructural analyses of the distribution of cell components in the apex of any tip-growing plant cell. The finding that almost every component had its own spatial arrangement demonstrates the complexity of the organization and regulation of the distribution of components in tip-growing cells.Abbreviations CER cisternal endoplasmic reticulum - ER endoplasmic reticulum - Nd numerical density - SE standard error - Sv surface density - TEM transmission electron microscopy - TER tubular endoplasmic reticulum - Vv volume fraction  相似文献   

19.
Summary Three different types of vesicles present before fertilization in the sea urchin egg were examined. The a-type corresponds to rough endoplasmic vesicles; the vesicles of b-type are rather smooth but may have vestigial granules within their membranes; the c-type belongs to the multivesicular bodies. Upon fertilization, vesicles appear in the outer cortical zone (vesicles of d-type). The majority of them arises by budding from the vesicles ofb-type. The budding occurs mainly at the basis of or within ridges of the cell surface; they may also be present in broader microvilli. The distance between the ridges was varied by early transfer of the eggs to calcium-free sea water. An inducing effect of the ridges on location of theb-vesicles and on the formation ofd-vesicles by budding could thus be demonstrated.Thed-vesicles appearing upon fertilization resemble in size and structure Golgi vesicles formed by budding from Golgi bodies present in the interior cortex or below it. Also theb-vesicles have their counterpart in the Golgi bodies. Theb andd-vesicles are therefore regarded as a Golgi system in which theb-vesicles represent dispersed Golgi bodies and thed- vesicles Golgi vesicles. Thed-vesicles may be designated as cortical Golgi vesicles, (c.G.v.).In thec-vesicles i.e. the multivesicular bodies (m.v.b.), a nucleid was observed which may be subdivided. A compartmentation of m.v.b. was observed which may lead to a detachment of vesicles of about the same size as thed-vesicles (c.G.v.) but probably of a different character.The differentiation of the fertilization membrane of the sea urchin egg occurs in two stages: the assembly and the solidification stage. The tentative conclusion is drawn that a secretion from the c.G.v. functions as an agent in the solidification process. The c.G.v. may also act upon the hyaline layer, both in its early formation and its maintenance during the course of embryonic development.The material for the present investigation was collected at Kristineberg Zoological Station and at Stazione Zoologica, Naples. The writer is indebted to the authorities of these stations for generous helop. Financial support was received from the Swedish Natural Sciences Research Council and the Swedish Cancer Society for which I express my gratitude.I also express my gratitude to Dr.Björn Afzelius for surveying the electron microscopic part of the work, to Dr.Jane Baxandall for permission to study her original electron micrographs and to Dr.Elsa Wicklund and Dr. L.Contoli for contribution to the experiments on the effect of calcium-free sea water. I thank Mrs.Astri Runnwström and Mrs.AnneMarie Ede for their able assistance.This research was carried out in association with the Research group of embryology for the study of cellular differentiation of the Department of Zoology, University of Rome (Director Professor P.Pasquini).  相似文献   

20.
Summary The ultrastructure of the ampullar epithelium of the human oviduct was studied in 9 women in different phases of the sexual cycle and in pregnancy.Both kinds of cells (secretory and ciliated) showed morphological signs of micropinocytotic activity during the follicular phase, but no such activity during the luteal phase. Thus cyclic variations were observed in both ciliated and secretory cells. The secretory cells were provided with an extraneous coat, which was most conspicuous during the follicular phase. The Golgi apparatus was large. The mitochondria had transversal cristae. The endoplasmic reticulum consisted of cisternae with partly granular membranes. The ribosomes were partly membranebound and partly free. The appearance of the endoplasmic reticulum and the distribution and frequency of the ribosomes varied with the functional states. The ciliated cells were not provided with an extraneous coat. The Golgi apparatus was small. The mitochondria were numerous and had transversal cristae. Multivesicular bodies and dense bodies occurred in the nuclear region. Cilia as well as centrioles were provided with striated rootlets.This investigation was supported by a grant from the foundation Therèse och Johan Andersson Minne.The skil ful technical assistance of Miss Gunilla Runefeldt is gratefully acknowledged.  相似文献   

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