Per Odin

Platelet-derived growth factor promotes survival of rat and human mesencephalic dopaminergic neurons in culture

SummaryThe effect of two isoforms of platelet-derived growth factor (PDGF), PDGF-AA and PDGF-BB, was tested on dissociated cell cultures of ventral mesencephalon from rat and human embryos. PDGF-BB but not PDGF-AA reduced the progressive loss of tyrosine hydroxylase- (TH)-positive neurons in rat and human cell cultures. The mean number of TH-positive cells in the PDGF-BB-treated rat culture was 64% and 106% higher than in the control cultures after 7 and 10 days in vitro, respectively. Corresponding figures for human TH-positive neurons were 90% and 145%. The influence of PDGF-BB was specific for TH-positive neurons and not a general trophic effect, since no change of either total cell number or metabolic activity was found. In PDGF-BB-treated cultures of human but not rat tissue the TH-positive neurons had longer neurites than observed in control or PDGF-AA-treated cultures. These data indicate that PDGF-BB may act as a trophic factor for mesencephalic dopaminergic neurons and suggest that administration of PDGF-BB could ameliorate degeneration and possibly promote axonal sprouting of these neurons in vivo.

Immunohistochemical localization of cellCAM 105 in rat tissues: appearance in epithelia, platelets, and granulocytes.

CellCAM 105 is an integral membrane glycoprotein, with apparent Mr 105,000, which has been purified from rat liver plasma membranes. It consists of two structurally similar, highly glycosylated polypeptide chains and is involved in cell-cell adhesion of adult rat hepatocytes in vitro. In this communication we report on the distribution and cell surface location of cellCAM 105 in rat tissues, obtained by using highly sensitive immunodetection systems based on complex formation between biotinylated antibodies, biotinylated peroxidase and avidin, or on antibodies coupled to alkaline phosphatase. CellCAM was found in many organs and organ systems, including liver, kidney, blood, blood vessels, glands, respiratory system, and gastrointestinal tract. It was mainly localized to epithelial structures but showed a varying cell surface distribution. In some cell types it was predominantly localized to cell-cell contact areas. In other cell types the highest concentrations were seen in brush-border areas containing densely packed microvilli. In addition to epithelial structures, cellCAM 105 was found in rat platelets, where it became strongly expressed on the cell surfaces after activation with ADP or collagen, suggesting that it might be involved in platelet adhesion and/or aggregation mechanisms. Granulocytes also contained cellCAM 105. By SDS-PAGE/immunoblotting, significant differences were found in the apparent Mr values of cellCAM 105 in different tissues. The collected data suggest that cellCAM 105 participates in several different cell surface membrane interactions, of which the common denominator might be membrane-membrane binding.

Dynamic expression of the cell adhesion molecule cell-CAM 105 in fetal and regenerating rat liver☆

Cell-CAM 105 is an integral cell surface glycoprotein that is involved in cell-cell adhesion of adult rat hepatocytes in vitro. In the present report we used a radio-immunoassay, a quantitative immunoblotting technique and immunofluorescence microscopy to investigate the expression of cell-CAM 105 in fetal and regenerating rat liver. In the fetal liver cell-CAM 105 did not appear until day 16 of the gestation, when it increased rapidly to reach the level found in adult liver, 3 weeks after birth. In liver regenerating after partial hepatectomy a transient decrease in the amount of cell-CAM 105 was observed in the plasma membranes of the hepatocytes. A significant decrease was observed as early as 12 h after partial hepatectomy, reaching a minimum by 3 days after the operation, corresponding to approx. 35% of the amount of cell-CAM 105 in normal liver. The amount then increased slowly and was back to the normal level by about 15 days after partial hepatectomy. The results indicate that cell-CAM 105 exerts its major function in terminally differentiated cells. An excellent correlation was seen between the kinetics of the expression of cell-CAM 105 and of reported changes of both enzymatic and organizational patterns of hepatocytes in regenerating and fetal liver. This suggests that cell-CAM 105 could be important for the development and maintenance of the cell-cell binding and organizational pattern characteristic of terminally differentiated hepatocytes.

Comparison of two cell-adhesion molecules, uvomorulin and cell-CAM 105.

Two cell adhesion molecules, cell-CAM 105 and uvomorulin (UM), were compared by analysing their antigenic structures, their activity in cell aggregation assays and their expression in various tissues. Cell-CAM 105 is a membrane glycoprotein which mediates the intercellular adhesion of reaggregating rat hepatocytes, and UM was first described to be involved in the compaction of preimplantation mouse embryos and embryonal carcinoma cells. UM is not only expressed during embryonic development but also in various adult tissues including liver, epithelia of lung, gut, kidney and uterus. A similar distribution for UM was found in rat tissues on cell types where cell-CAM 105 is known to be present. Our studies show that (i) cell-CAM 105 and UM are distinct and different proteins; (ii) uvomorulin is involved in the compaction of rat preimplantation embryos but Fab anti-UM has no effect on reaggregating rat hepatocytes, where Fab anti-cell CAM is effective; (iii) distribution studies show that UM is expressed on a broader range of epithelial cells while cell-CAM 105 is more restricted to hepatocytes and simple epithelia. In cases where both cell adhesion molecules are expressed on the same cell types they can be localized to different parts of the cell surface.

Quantitative determination of the organ distribution of the cell adhesion molecule cell-CAM 105 by radioimmunoassay

We have previously identified a 105,000-Da plasma membrane glycoprotein, denoted cell-CAM 105, that is involved in intercellular adhesion of reaggregating rat hepatocytes. In this communication we report on the development of a radioimmunoassay for cell-CAM 105, employing purified cell-CAM 105, specific antisera against the molecule, and formalin-fixed protein A-containing staphylococci for precipitation of the immune complexes. The assay was shown to be sensitive, specific, precise, rapid, and easy to perform. We used this radioimmunoassay in investigations of the occurrence of cell-CAM 105 in different rat organs. Cell-CAM 105 was present in a wide spectrum of organs in varying amounts. The highest concentrations were found in the gastrointestinal tract, liver, some secretory glands, vagina, kidney, and lung. In addition, cell-CAM 105 was detected in blood, where it was shown to reside mainly in the platelets. Other tissues, particularly the central nervous system and muscle tissues, were cell-CAM-negative. The results were confirmed by immunoblotting, which revealed one distinct protein component, corresponding to cell-CAM 105, in each positive organ.

Two different cell adhesion molecules—Cell-CAM 105 and a calcium-dependent protein—Occur on the surface of rat hepatocytes☆

We have recently identified a 105000 D plasma membrane glycoprotein, denoted cell-CAM 105 (CAM, cell adhesion molecule), that is involved in intercellular adhesion of reaggregating rat hepatocytes (Ocklind & Obrink, J biol chem 257 (1982) 6788 [11]). In this communication we identify another cell surface protein that is also involved in hepatocyte cell-cell adhesion. This protein has an apparent molecular weight (MW) of 70000 and can be released from the surface membrane by chelation of calcium with EGTA. Results are presented indicating that it is identical with a previously discovered protein, CDP-1 (CDP, calcium-dependent protein) (Obrink, Lindström & Svennung, FEBS lett 70 (1976) 28 [28]). Antisera produced against either cell-CAM 105 or CDP-1 inhibit hepatocyte aggregation, but not attachment to collagen. Cell-CAM 105 and CDP-1 are present on the cell surface as separate components, as judged by the fact that both EGTA treatment and trypsin treatment of hepatocytes selectively make the cells insensitive to blocking of aggregation by antibodies against CDP-1 but not by antibodies against cell-CAM 105. However, although much less efficiently, the antibodies against CDP-1 can recognize a 105000 D protein which is also bound by the antibodies against cell-CAM 105, and under certain conditions the antibodies against cell-CAM 105 seem to recognize a 70000 D protein. CDP-1 may thus be derived from cell-CAM 105, or the two proteins might have a common precursor.

The cell-surface expression of the cell adhesion molecule cellCAM 105 in rat fetal tissues and regenerating liver☆

In the present investigation we have used a sensitive immunohistochemical technique to study the appearance and cell-surface distribution of cellCAM 105 in rat fetal tissues and in regenerating liver. CellCAM 105 is an integral membrane glycoprotein that is involved in cell-cell adhesion of mature rat hepatocytes in vitro. In 12-day-old rat fetuses no cellCAM 105 was detected. CellCAM 105 then appeared on Day 13 in megakaryocytes of the fetal liver, on Day 16 in the liver parenchyme, and on Day 17 in the epithelial cells of the proximal kidney tubules and of the small intestinal mucosa. In the liver parenchyme cellCAM 105 first appeared in immature bile canaliculi. During Days 19-21 a significant staining also occurred on the contiguous sides of the hepatocytes, which at that time became closely associated when the blood-forming cells disappeared. This surface staining then gradually disappeared and 2-3 weeks after birth cellCAM 105 was expressed in the bile canalicular area which is typical of mature hepatocytes. In regenerating liver the amount of cellCAM 105 decreases to a minimum 2-3 days post-hepatectomy, then increases and reaches the normal concentration 10-15 days post-hepatectomy [Odin and Obrink (1986) Exp. Cell Res. 164, 103-114]. The cell-surface distribution of cellCAM 105 also changed, and on Days 3-5 post-hepatectomy it appeared on all faces of the hepatocytes which then were closely associated without obvious sinusoids in between. This staining pattern then slowly changed toward the normal pattern of mature liver, which appeared about 15 days post-hepatectomy. A theoretical analysis of the mode of hepatocyte cell division during liver regeneration suggested that the surface of the postmitotic hepatocytes should become unpolarized with respect to macromolecular composition. This is in agreement with the observed surface distribution of cellCAM 105. The results support the hypothesis that cell-surface interactions mediated by cellCAM 105 might contribute to the regular organization of hepatocytes in the normal, mature liver plates.

Platelet-derived growth factor receptor expression after neural grafting in a rat model of Parkinson's disease

Platelet-derived growth factor (PDGF) has trophic effect on dopaminergic neurons in vitro. We have previously shown dynamic changes in the expression of PDGF in embryonic mesencephalic grafts and surrounding host striatal tissue following intracerebral transplantation in a rat model of Parkinsons disease. In this study the expression of the PDGF receptors was examined in the same model using immunohistochemistry. Most ventral mesencephalic (VM) cells from E13–E15 rat embryos possessed both PDGF α-and β-receptors before implantation. Double immunofluorescence staining revealed that about 10% of the cells also expressed tyrosine hydroxylase (TH). The PDGF α-receptor was detectable in the graft up to 1 wk after transplantation but had disappeared at 3 wk. In the host tissue, scattered glial cells were positive for the α-receptor but the expression was unchanged following transplantation. The β-receptor expression almost completely disappeared from the grafted tissue by 4 h following transplantation, and only a few cells of the host striatum showed immunoreactivity. However, after 3 wk β-receptor positive cells were again detectable in the graft. These cells appeared to be endothelial cells as identified by an antibody against von Willebrands factor. Our data suggest that PDGF might act locally on embryonic dopaminergic cells in an autocrine or juxtacrine manner before and shortly after transplantation, and on surrounding glial cells in a paracrine manner after transplantation. Furthermore, PDGF-BB might influence neovascularization in the graft.

Comparison and functional characterization of C-CAM, glycoprotein IIb/IIIa and integrin β1 in rat platelets

Three adhesion-related proteins in rat platelets were compared, namely C-CAM, gpIIb/IIIa, and integrin beta 1. These three proteins behaved as distinct components as judged by both biochemical and immunological analyses. GpIIb/IIIa bound to a GRGDSPC-peptide, but neither beta 1-containing integrins nor C-CAM had any affinity either for this peptide or for a large cell-binding fragment of fibronectin. C-CAM and integrin beta 1 behaved differently when platelets were labeled with 125I, solubilized by detergent, and immunoprecipitated. Significant amounts of labeled C-CAM was precipitated when the platelets were first solubilized with detergent and then 125I-labeled. Almost no labeled C-CAM could be precipitated when intact platelets, that were unactivated or activated by ADP, were labeled. In contrast, labeled integrin beta 1 was immunoprecipitated when unactivated platelets were surface-labeled. However, when platelets that were activated by ADP and calcium ions were labeled almost no labeled integrin beta 1 could be immunoprecipitated. These data indicate 1) that C-CAM in intact platelets is inaccessible to surface-labeling and 2) that beta 1 integrin is less accessible to surface-labeling after platelet activation in the presence of calcium ions.