共查询到20条相似文献,搜索用时 31 毫秒
1.
It is found that hepatic cells of intact rats measuring 129-192 mcm2 are resistant to cytotoxical action of carbon tetrachloride (CCl4). After a single interperitoneal injection of the carcinogen 4-dimethylaminoazobenzene, small hepatocytes (64-128 mcm2) appear to be maintained for one month following five injections of phenobarbital. These small hepatocytes are resistant to cytotoxical action of CCl4. The resistance was studied using a cytochemical test on succinate dehydrogenase. A direct dependence exists between the cell size and the sensitivity to CCl4 among large sized hepatocytes. 相似文献
2.
Adult rat hepatocytes were cultured for 15 days on type I collagen-coated permeable membranes in a hormonally defined Waxman's modified medium supplemented with very low concentrations of insulin, glucagon and dexamethasone. Phase contrast examination showed that 15-day-old cultures still formed a regular monolayer of polygonal cells. In similarly aged cultures, intracellular glycogen was abundant and evenly distributed, while steatosis remained very limited. Scanning and transmission electron microscopy showed that well developed bile canaliculi could be observed on the lateral side of the hepatocyte membrane after 4 days of incubation and persisted for 2 weeks. These canalicular structures probably originated from coalescence of membrane invaginations observed in 1-day-old cultures. Transmission electron microscopy showed that the ultrastructure of the cells was very close to that of normal rat hepatocytes in the intact liver. These results suggest that rat hepatocytes cultured under these experimental conditions are able to develop and maintain tissue-specific cytochemical and morphological properties for at least 15 days. 相似文献
3.
Prenatal exposure to alcohol alters the Golgi apparatus of newborn rat hepatocytes: a cytochemical study 总被引:7,自引:0,他引:7
J Renau-Piqueras F Miragall C Guerri R Baguena-Cervellera 《The journal of histochemistry and cytochemistry》1987,35(2):221-228
The effect of prenatal exposure to ethanol on the Golgi apparatus of newborn rat hepatocytes has been studied cytochemically using several trans-Golgi markers (thiamine pyrophosphatase, uridine diphosphatase, inosine diphosphatase, acid phosphatase, and 5'-nucleotidase) as well as a cis-side marker (osmium impregnation). The amount of cerium phosphate formed in the cytochemical reactions was roughly quantitated by stereologic methods. The Golgi apparatus of about 40% of the hepatocytes appeared disorganized after alcohol treatment, and in the other 60%, the electron density of reaction product deposits for all phosphatases investigated was decreased. 5'-Nucleotidase was completely absent in cisternae of Golgi apparatus of treated cells. In control cells impregnated with osmium tetroxide, reduced osmium compounds were observed in most Golgi cisternae and in nearby vesicles. In contrast, only small vesicles appeared positive in treated hepatocytes. These results suggest that prenatal alcohol exposure alters some Golgi functions. Thus, the decrease in nucleoside diphosphatase and 5'-nucleotidase cytochemical activities after ethanol exposure strongly suggests that this treatment could affect glycosylation in the Golgi apparatus of newborn rat hepatocytes. 相似文献
4.
J J Trout J M Viles 《Virchows Archiv. B, Cell pathology including molecular pathology》1978,28(3):279-285
Electron microscopic morphological and cytochemical techniques were used to follow the sub-cellular events that accompanied Triton WR-1339 accumulation in hepatocytes. Localization of two lysosomal enzymes, acid phosphatase and aryl sulfatase, clearly established the lysosomal nature of the Triton WR-1339 containing electron-lucid structures that appear in hepatocytes following treatment with this compound. 相似文献
5.
V V Ul'ianova L V Molostova A S Loginov 《Biulleten' eksperimental'no? biologii i meditsiny》1990,109(5):510-512
The deposits of cholesterol in hepatocytes in alcoholic rats were studied by means of cytochemical method. It was shown the localization of cholesterol in intracellular as well as extracellular compartment. In numerous hepatocytes there was observed a destruction of mitochondria, rough endoplasmic reticulum and plasma membranes. It was observed two structural forms of digitonin cholesterol complexes: structures like "crinkles" and small cylinders and multilamellar ones. 相似文献
6.
F De Paermentier 《Comptes rendus des séances de la Société de biologie et de ses filiales》1978,172(6):1242-1244
The effects of an extract from adult liver on chick embryo hepatocytes and fibroblasts cultivated in vitro are studied with cytological and quantitative cytochemical methods. This substance inhibits the multiplication of hepatocytes adn prevents these cells from entering into the S phase of the cycle. No effect is observed on the chick embryo fibroblasts. Not any morphological alterations are observed in the two cell types. 相似文献
7.
K Winzer C J Van Noorden A K?hler 《The journal of histochemistry and cytochemistry》2001,49(8):1025-1032
There is a great need for rapid but reliable assays to determine quantitatively effects of xenobiotics on biological systems in environmental research. Hepatocytes of European flounder are sensitive to low-dose toxic stress. Glucose-6-phosphate dehydrogenase (G6PDH) is the major source of NADPH in cells and is therefore of major importance for NADPH-dependent xenobiotic biotransformation and defense against toxic injury. These facts prompted us to develop a sensitive cytochemical method to detect G6PDH activity in living isolated flounder hepatocytes using the tetrazolium salt method. The intact plasma membrane did not appear to be a barrier for substrate, co-enzyme, and dye molecules because the intracellular enzyme reaction started immediately when incubation medium was added and could be monitored in real time per individual cell using image analysis. The reaction was effectively stopped for end point measurements by using 4% formaldehyde in 0.1 M phosphate buffer (pH 5.3). The final reaction product, formazan, was stable in hepatocytes for at least 12 days at 4C. This is the first time that a chromogenic histochemical assay is applied to living cells. This approach provides an easy tool for large-scale screening of xenobiotic metabolism and cellular stress defense. 相似文献
8.
Dr. A. V. LeBouton 《Cell and tissue research》1982,227(2):423-427
Summary The indirect immunoperoxidase method was used to identify albumin in hepatocytes of rats before and after periods of starvation. All hepatocytes in fed rats contained a relatively large amount of nascent albumin. Overnight fasting reduced the number of hepatocytes with a large amount of albumin to primarily those surrounding terminal hepatic venules. These were estimated to be about 30% of the population. The other cells had only a slight amount of albumin. After 48 h of fasting all hepatocytes contained a low level of albumin. 相似文献
9.
A new method for enzyme cytochemical studies on individual cells is developed. Cells are incorporated in the matrix of a thin film of transparent polyacrylamide prior to incubation in a cytochemical medium. Five different kinds of individual cells, i.e. isolated rat hepatocytes, isolated mouse oocytes, cultivated human fibroblasts, rat thymocytes and human blood cells are used for testing the applicability of this method for the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetranitro BT. The incorporation technique solves at least some of the problems occurring with enzyme cytochemistry on single cells. The morphology of the cell is very well preserved, the formazan precipitation due to enzyme activity occurs entirely within the cell cytoplasm, the nothing dehydrogenase activity can be kept very low and the loss of cells is completely prevented with all cell types used. 相似文献
10.
To optimize conditions for retroviral-mediated transfer of a recombinant gene to hepatocytes, the pMNSM-Tk-lacZ vector, which we had constructed to express the bacterial beta-galactosidase gene, was transduced to rat hepatocytes under various conditions, and the expression of beta-galactosidase activity was examined by cytochemical staining. Compared to the hepatocytes of adult rats, those of newborns were about 50-100 times more sensitive to transduction with the beta-galactosidase gene in vitro. The sensitivity was high in the newborn hepatocytes when the virus was infected on days 1 and 2 after initiation of culture. However, the sensitivity to infection did not correlate with the DNA synthetic activity. The gene transfer was feasible not only to hepatocytes in monolayer culture but also to those in spheroid culture. The spheroidal aggregates containing hepatocytes transduced with the beta-galactosidase gene could be transplanted into the spleen of syngenic adult rat, although the expression was very low. 相似文献
11.
P B Gahan J Middleton 《Comparative biochemistry and physiology. A, Comparative physiology》1982,71(2):345-348
1. Quantitative cytochemical studies of hepatocytes from a variety of species representing the various sub-classes of mammals have shown that euploidization of hepatocytes is a typical feature of mammalian livers. 2. Euploidization occurs during early development and frequently involves a large proportion of the hepatocyte population. 3. Comparison with livers from birds and diploid fish indicates that euploidization is a special mammalian feature. 相似文献
12.
The authors studied the deposits of cholesterol in the liver of alcoholic men and rats by means of cytochemical method. The localization of cholesterol in intracellular and extracellular compartments was shown. In numerous hepatocytes there were focal destructions in mitochondria, rough endoplasmic reticulum and plasma membranes. 相似文献
13.
Thomas L. Jetton Masa Shiota Susan M. Knobel David W. Piston Alan D. Cherrington Mark A. Magnuson 《Experimental diabetes research》2001,2(3):173-186
Hepatic glucokinase (GK) is acutely regulated by
binding to its nuclear-anchored regulatory protein
(GKRP). Although GK release by GKRP is tightly
coupled to the rate of glycogen synthesis, the nature
of this association is obscure. To gain insight into
this coupling mechanism under physiological stimulating
conditions in primary rat hepatocytes, we
analyzed the subcellular distribution of GK and
GKRP with immunofluorescence, and glycogen deposition
with glycogen cytochemical fluorescence,
using confocal microscopyand quantitative image
analysis. Following stimulation, a fraction of the GK
signal translocated from the nucleus to the cytoplasm.
The reduction in the nuclear to cytoplasmic
ratio of GK, an index of nuclear export, correlated
with a >50% increase in glycogen cytochemical
fluorescence over a 60min stimulation period. Furthermore,
glycogen accumulation was initially deposited
in a peripheral pattern in hepatocytes
similar to that of GK. These data suggest that a compartmentalization
exists of both active GK and the
initial sites of glycogen deposition at the hepatocyte
surface. 相似文献
14.
Plasma lipoproteins (and other ligands) are endocytosed by hepatocytes and appear in multivesicular bodies (MVBs) in the Golgi-lysosome region of the cell prior to their degradation. We have isolated MVB fractions from livers of estradiol-treated rats, permitting studies of their properties (Hornick et al. 1985). Here we report our cytochemical studies of lysosomal enzyme activity in partially and highly purified MVB fractions and in MVBs in hepatocytes in situ. Only about 15% of partially or highly purified MVBs were positive for acid phosphatase and arylsulfatase, consistent with the prelysosomal nature of this compartment. Partially purified MVB fractions contained small round vesicles, 70-120 nm in diameter, which stained intensely for these enzymes; occasionally these vesicles appeared to fuse with MVBs, suggesting that these structures are primary lysosomes. Such stained vesicles were rarely seen in highly purified MVB preparations. Acid phosphatase reaction product with cerium as capture reagent appeared as uniform precipitates surrounding endocytosed plasma lipoproteins in positively stained MVBs. Arylsulfatase reaction product, however, appeared as distinctive arc or plaque-like deposits just inside the MVB-limiting membrane, often in continuity with intense reaction product contained in a fusing primary lysosome. Similar putative primary lysosomes were occasionally observed in isolated, "intact" Golgi fractions from the same livers. Similar histochemical reactivities of MVBs and putative primary lysosomes were observed in thin sections of hepatocytes in situ. These observations support the conclusion that, in hepatocytes, MVBs represent the immediate prelysosomal compartment in the endocytic pathway of macromolecular catabolism, and suggest that MVBs are converted to secondary lysosomes by direct fusion with primary lysosomes arising from closely adjacent Golgi compartments. 相似文献
15.
Plasma membrane specialization and intracellular polarity of freshly isolated rat hepatocytes 总被引:2,自引:0,他引:2
G M Groothuis C E Hulstaert D Kalicharan M J Hardonk 《European journal of cell biology》1981,26(1):43-51
It was investigated whether rat hepatocytes maintain their plasma membrane specialization (sinusoidal, lateral and bile canalicular sites) and their intracellular polarity (peribiliary region, rich in lysosomes and poor in mitochondria) after isolation. The morphology of the hepatocytes and the cytochemical localization of marker enzymes for the bile canalicular membrane (alkaline phosphatase, adenosine triphosphatase and 5' nucleotidase), for the lysosomes (acid phosphatase) and for the mitochondria (beta-hydroxybutyrate dehydrogenase and succinate dehydrogenase) were studied in situ and directly after isolation using both light and electron microscopy. The morphology of the cells and the cytochemical activity of acid phosphatase, succinate dehydrogenase and beta-hydroxybutyrate dehydrogenase showed that in isolated cells, as in situ, the lysosomes were concentrated in bands, devoid of mitochondria. Unlike in situ the reaction product of alkaline phosphatase, adenosine triphosphatase and 5'nucleotidase was evenly distributed along the entire plasma membrane of the isolated cells. Morphologically, no tight or gap junctions or desmosomes could be detected in the isolated cells, while the plasma membrane appeared to be homogeneously covered with uniform microvilli. In conclusion it can be stated that during isolation the hepatocytes loose their distinct plasma membrane specialization, but maintain their peribiliary region rich in lysosomes and poor in mitochondria. 相似文献
16.
F de Paermentier 《Comptes rendus des séances de la Société de biologie et de ses filiales》1978,172(3):565-568
The effects of an extract from adult liver on hepatoma cells cultivated in vitro (line H-35) are studied with cytological and quantitative cytochemical methods. This extract inhibits cell multiplication and premitotic DNA synthesis. The treated cells form "trabeculae" looking like those observed when normal adult hepatocytes are cultivated in vitro. 相似文献
17.
C. J. F. van Noorden J. Tas I. M. C. Vogels G. G. de Schepper 《Histochemistry and cell biology》1982,74(2):171-181
Summary A new method for enzyme cytochemical studies on individual cells is developed. Cells are incorporated in the matrix of a thin film of transparent polyacrylamide prior to incubation in a cytochemical medium. Five different kinds of individual cells, i.e. isolated rat hepatocytes, isolated mouse oocytes, cultivated human fibroblasts, rat thymocytes and human blood cells are used for testing the applicability of this method for the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetranitro BT. The incorporation technique solves at least some of the problems occurring with enzyme cytochemistry on single cells. The morphology of the cells is very well preserved, the formazan precipitation due to enzyme activity occurs entirely within the cell cytoplasm, the nothing dehydrogenase activity can be kept very low and the loss of cells is completely prevented with all cell types used. 相似文献
18.
A quantitative cytochemical method was developed for measuring the GSH (reduced glutathione) content of hepatocytes in different regions of the rat liver lobule. Use of this method enabled us to show that GSH is not evenly distributed within the rat liver lobule. The hepatocytes located within 100 micrometer of the central vein contain much less GSH than do those in other regions of the rat liver lobule. We suggest that this partially explains the peculiar susceptibility of these cells to electrophilic attack by toxic metabolites formed via the microsomal cytochrome P-450 system. 相似文献
19.
酶是参与植物体内生化反应的特殊蛋白质。在保持活组织和细胞结构完整性的条件下,利用组织化学、细胞化学、免疫学和显微检测等技术研究酶的即位定位,是了解酶在组织、细胞和亚细胞中的分布、活性动态与定量及酶功能等的重要途径。对植物体中酶定位的组织化学和细胞化学方法的概念、原理与研究进展进行了综述,并根据国际酶化学分类编号顺序,分别介绍了25种酶的组织化学染色定位所用的反应介质和染色方法及46种酶的细胞化学定位方法的参考文献。 相似文献
20.
K Furukawa Y Mochizuki N Sawada M Gotoh H Tsukada 《Virchows Archiv. B, Cell pathology including molecular pathology》1988,55(5):279-285
The present ultrastructural morphometric and cytochemical studies demonstrate clofibrate induced changes in peroxisomes in adult rat hepatocytes maintained for 14 days in primary culture on floating collagen gels. Catalase activity and the number and diameter of peroxisomes were reduced in hepatocytes cultured for between 2/3 and 7 days. However, hepatocytes cultured for 7-14 days had well-developed peroxisomes containing crystalloid nucleoids. The number of anucleoid peroxisomes in hepatocytes treated with 2 mM Na clofibrate increased with culture age, and by day 14 the number was 2.9 times greater than in freshly isolated hepatocytes. Catalase activity, as well as the number of nucleoid-containing peroxisomes were much greater in treated hepatocytes than in untreated controls, but decreased slightly with culture age. The diameter of peroxisomes was not reduced in the treated cells. These results suggest that the treatment with Na clofibrate is effective both for proliferation and maintenance of peroxisomes and for enhancing catalase activity. In treated hepatocytes, matrical plates were formed in peroxisomes from days 5 to 14 and the number of plate-containing peroxisomes increased with culture age. 相似文献