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Mehr von diesem Entwickler Alle anzeigen. LiveScore: Fussball Welt Das gefällt dir vielleicht auch Alle anzeigen. At this low magnification, intercellular occluding junctions are not visible.
Nuclei of hepatocytes are indicated HN. D: Higher magnification electron micrograph illustrating a canaliculus, and adjacent occluding junctions arrows.
Adjacent hepatocytes are separated by an intercellular space with occasional intercellular junctional complexes.
The intercellular space widens to form canaliculi fig. Golgi apparatus was seen occasionally in the cytoplasm adjacent to the canaliculi.
Non-parenchymal cells were also detected with electron microscopic methods, as illustrated in figure 7. Kupffer cells were encountered frequently, either situated upon underlying endothelial cells or as part of the lining of the sinusoidal capillaries.
Figure 7A presents an electron micrograph of tissue that had been processed immunocytochemically for F The immunoreactivity is visible as dark staining on the plasmalemma, which displays folds and pseudopodia.
Kupffer cells display large numbers of lysomes, several of which can be seen in the cell in figure 7A.
Kupffer cells are also shown in figure 7D , where adjacent membranes between Kupffer and endothelial cells display no specialized contacts, and in figure 7G , where uptake of gold labeled liposomes is demonstrated by the many gold particles in the Kupffer cell cytoplasm, but not in the nucleus.
Electron micrographs of non-parenchymal cells. A: Electron micrograph of a section processed immunocytochemically for F, showing cell surface immunoreactivity black arrows of a Kupffer cell K ; the nucleus of the Kupffer cell KN is visible.
White arrows indicate lysomes within the Kupffer cell. The Kupffer cell is situated in the sinusoidal lumen Lu , above an hepatocyte H.
B: Micrograph showing an Ito stellate cell I situated between two hepatocytes H and an endothelial cell E. The Ito cell is characterized by cytoplasmic lipid droplets LD.
C: High magnification electron micrograph showing the space of Disse D situated under fenestrated F endothelial processes EP above an hepatocyte H and below the capillary lumen Lu.
Also seen is a putative Ito cell process IP. D: Close membrane association, but without specialized membrane attachments, between a Kupffer cell K and an endothelial cell E.
E: Adhering junctions arrows between processes of 2 endothelial cells E , overlying the space of Disse D and an hepatocyte H.
F: Pit cell with nucleus PN situated in a sinusoidal lumen, adjacent to an endothelial cell E. G: Micrograph showing gold labeled particles associated with engulfed liposomes in a Kupffer cell Kupffer cell nucleus KN is indicated.
Ito stellate cells could be identified by the presence of prominent intracellular lipid droplets and filamentous material, as illustrated in figure 7B.
These stellate cells were situated between endothelial cells and hepatocytes or were intercalated between hepatocytes as in figure 7B.
Endothelial cells were identified by their elongated and flattened nuclei, and by the presence of fenestrations Fig. These fenestrations have diameters of approximately to nm.
Fenestrations did not include membranous diaphragms over the openings; only sparse evidence of a very rudimentary basal lamina was seen associated with endothelial cells.
At sites where cell processes from apparent adjacent endothelial cells meet, specialized adhesive intercellular junctional complexes are seen Fig.
Adhesive contacts are not seen between endothelial cells and hepatocytes or any other non-parenchymal cell. Finally, pit cells are occasionally seen, as illustrated in figure 7F.
The data presented here provide an overview of the histological and fine structural organization of the mature mouse liver.
It is clear that the fundamental features of the mouse liver are remarkably similar to the features of liver from other mammalian species.
On the one hand, the clear similarities may detract from the noteworthiness of this manuscript; the mouse liver displays the same cellular and subcellular constituents as do the livers of other mammalian species.
On the other hand, however, the data presented in this manuscript are vitally important for the interpretation and understanding of other results, using either wild type or genetically engineered mice, and provide a solid reference point against which other data can be interpreted.
Although the very sparse connective tissue within the mouse liver makes recognition of its lobular structure e.
The types of cells that comprise the mouse liver are similar to those that have been described in other mammalian species. The prominent cell type is the hepatocyte, characterized by the presence of intracellular albumin Bernuau et al.
Also studied were the Ito stellate cells Fahimi, ; Gard et al. As with any organ, endothelial cells form much of the lining of the sinusoidal capillaries.
The relative proportions of these cells are similar to figures reported for other species Bouwens et al.
Further, the biliary system of the mouse liver appears similar to that of other mammalian species, with the presence of canaliculi between adjacent hepatocytes and small bile ductules that form part of portal areas.
Thus, although the present investigation did not reveal novel features of liver organization in the mouse, the value of this study is that it establishes baseline data for the mouse and demonstrates the similarities of the histological organization of the mouse liver and of other mammalian species.
These data underscore the value of the mouse model for laboratory studies of liver structure and function. Different cell types in the liver were identified based upon their morphological features and by their immunoreactivity.
The present results from mouse are fully compatible with these previous reports. The present results indicate that albumin positive hepatocytes are encountered approximately three times more frequently than are Kupffer cells, and about six to seven times more frequently than Ito stellate cells.
Further, the present studies demonstrate that hepatocytes also contain biotin. Biotin appears not to be synthesized by hepatocytes but rather is taken up from food or produced by intestinal flora Bowman and Russell, , and is stored in the liver to support its role in metabolism.
The presence of biotin within hepatocytes clearly is important for considerations of its metabolic role, but may also have importance in issues of immunocytochemical methodology.
Immunocytochemical studies that use avidin-biotin binding as a step in localizing some antigen of interest may yield false positive results due to avidin binding to endogenous biotin rather than to biotinylated secondary antibodies Ramos-Vara, The fine structural features of mouse hepatocytes are similar to those reported for other mammalian species.
Large round nuclei, and occasionally two nuclei, are prominent. Much of the cytoplasm is occupied by mitochodria, both smooth and rough endoplasmic reticulum, and scattered glycogen particles.
The basolateral portion of the hepatocyte, which faces the sinusoidal capillaries, displays a rich microvillous elaboration of the plasmalemma; this microvillous border occupies much of the space of Disse.
As in other mammalian species, the apical portion of hepatocytes is associated with canaliculi. Canaliculi are formed by a widening of the intercellular space, although the canalicular lumens are set off from the intercellular space by occluding junctions.
Small profiles of Golgi apparatus were seen occasionally in the apical cytoplasm close to canaliculi, but we did not detect secretory vesicles associated with canaliculi.
The canaliculi are believed to form an anastomosing system, which lead eventually through ducts of Hering into small bile ductules in the portal areas.
These small bile ducts are a consistent feature in the portal areas. Monocyte derived macrophages are found in virtually every organ and tissue of the body, and comprise the diffuse reticulo-endothelial system Aschoff, ; Furth et al.
In the liver these macrophages are termed Kupffer cells von Kupffer, ; Widmann et al. Although originally, these phagocytic cells were likely confused with the stellate cells that would later be identified by Ito , later studies demonstrated that Kupffer cells can be identified by their ability to phagocytose tracer substances, including carbon, India ink, or latex microspheres, and also by their immunoreactivity to the F antibody.
The present studies demonstrated the presence of F positive Kupffer cells in mouse liver, and electron microscopic studies demonstrated this antibody labels a cell surface marker.
Further, the results from studies of double labeling with fluorescently labeled latex microspheres and also from immunocytochemistry have demonstrated conclusively that the Kupffer cells are a population of cells distinct from the Ito stellate fat storing cells.
The use of latex microspheres of different diameters was useful in demonstrating that Kupffer cells could be labeled specifically with larger 0.
Other investigators Bouwens et al. These authors Sleyster and Knook, ; Bouwens et al. The present data corroborate this finding in the mouse, although the regional differences in the mouse liver appear not as great as the regional differences reported for rat liver.
Ito stellate cells are fat storing cells of the liver, and in the present studies these were identified by immunoreactivity to glial fibrillary acidic protein GFAP Gard et al.
Stellate cells are identifiable by their fine structural features of prominent intracellular lipid droplets and by cytoplasmic filamentous material.
The intracellular filamentous material likely forms the basis of their immunoreativity to GFAP and to desmin Leo and Lieber, ; Yokoi et al.
Quantitative estimates Leo and Lieber, ; Yokoi et al. Further, de Bleser et al. The present study noted that stellate cells were not distributed homogenously throughout the liver, but a consistent pattern between peri-portal and peri-central regions was not detected.
Endothelial cells are an important cell type in any organ, and certainly so in the liver. The very sparse nature of a basal lamina beneath the endothelial cells, along with the absence of diaphragmatic coverings of the fenestrations, allows for apparent relatively free movement of small molecules less than nm diameter between the space of Disse and the capillary lumen.
Wisse has demonstrated both small bristle-coated micropinocytotic vesicles and large smooth macropinocytotic vesicles in the endothelial cells.
Small latex microspheres, diameter of 30 nm, are not detected in hepatocytes after intravascular injection, although they do appear to label endothelial cells.
This suggests that the latex microspheres are taken up by endothelial vesicles, but either do not reach the space of Disse or are not taken up by the hepatocyte microvillous border in the space of Disse.
The present analysis indicated that a considerable portion of hepatocytes are double nucleated.
The percentage gleaned from studies of tissue slices is likely to be an underestimate, however, because although two nuclei in the X or Y planes would be detected easily, if a second nucleus was obscured in the Z plane or found within an adjacent section, it would be missed by the techniques employed in this study.
Another approach taken in the present studies was to dissociate liver cells, and use the Trypan blue stain to detect double nucleated cells.
These results are in agreement with other recent studies Wheatley, ; Gupta, that have reported that the population of hepatocytes includes many double nucleated cells.
The functional significance of double nucleated cells, however, is not clear. No evidence was revealed to indicate that any other cell type within the liver included double nucleated cells.
Kupffer and stellate cells probably were not detected in the dissociated cell studies, as different steps in the cell dissociation process need to be taken to include these cell populations Smesdrod et tal.
The thin squamous endothelial cells display extensive spread of cytoplasm and membranes, making the detection of two nuclei within one cell a very difficult endeavor.
We are not aware of any examples of double nucleated endothelial cells, and believe they are not likely to exist.
Genetically engineered mice will play a very important role in future studies of liver function, and so it is vitally important to have baseline reference information on the cellular makeup of normal mouse liver.
The present paper, using histological, quantitative immunocytochemical, and fine structural analyses, demonstrates that the cellular organization of the mouse liver is quite similar to that of other mammalian species, confirming that the mouse presents a useful animal model for studies of liver structure and function.
National Center for Biotechnology Information , U. Histochem Cell Biol. Author manuscript; available in PMC Oct Janie L.
Longmuir , 2 and Richard T. Robertson 1. Kenneth J. Richard T. Author information Copyright and License information Disclaimer. Address for correspondence: Richard T.
Robertson, Ph. Copyright notice. The publisher's final edited version of this article is available at Histochem Cell Biol. See other articles in PMC that cite the published article.
Abstract The cellular organization of normal mouse liver was studied using light and electron microscopy and quantitative immunocytochemical techniques.
Introduction The important roles performed by the liver, not only in the storage and release of nutrients but also in the neutralization and elimination of a variety of toxic substances, have prompted investigations of its cellular constituents and organization.
Lectin Binding Cryotstat cut sections were mounted on slides and rinsed in Tris buffer. Hematoxylin and Eosin Slide mounted 8—12 um cryostat sections were dehydrated through absolute alcohol and rehydrated to water.
Electron Microscopy Liver tissue from 9 animals was used for electron microscopic studies.
Dissociated Cell Staining Mice aged 30 days to adult were deeply anesthetized with sodium pentobarbital and perfused through the portal vein with 25 ml cold 0.
Quantitative Assessments Relative numbers of each of the different populations of cells were estimated by counting the number of nuclei of immunocytochemically labeled cells and correcting the raw counts by using the method of Abercrombie Open in a separate window.
Further Immunocytochemical Characterization of Liver Cells The issue of whether the immunocytochemical markers resulted in overlapping populations of cells was addressed.
Double Nucleated Cells The issue of determining whether these cells were double nucleated was addressed by studying immunocytochemically identified cell types along with DAPI staining of cell nuclei.
Fine Structural Features of Liver Tissue An analysis of the fine structural features of hepatocytes and non-parenchymal cells was undertaken using electron microscopic techniques.
Discussion Overview The data presented here provide an overview of the histological and fine structural organization of the mature mouse liver.
Double Nucleated Cells The present analysis indicated that a considerable portion of hepatocytes are double nucleated.
Conclusion Genetically engineered mice will play a very important role in future studies of liver function, and so it is vitally important to have baseline reference information on the cellular makeup of normal mouse liver.
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