Expression and distribution of GAP‐43 in human astrocytes in culture By combining mRNA analysis and immunocytochemistry, we investigated the expression of the growth associated protein 43 (GAP‐43) in enriched populations of astrocytes, obtained from mixed cultures of human fetal brains. Total cellular RNA was extracted from cell pellets and reverse transcribed into cDNA; cDNA was subjected to PCR amplification using primers specific for GAP‐43 and PCR products were separated through polyacrylamide gels. Double immunofluorescence staining was performed on dissociated cell cultures using antibodies to glial fibrillary acidic protein (GFAP) and to GAP‐43. Results showed that both transcription and translation for GAP‐43 occur in cultured astrocytes. GAP‐43 immunoreacting material was detected in the cell processes and diffusely in the cytoplasm of GFAP‐positive astrocytes, during early stages of maintenance in vitro. In older cultures, GAP‐43 immunoreactivity persisted in a large percentage of cells, with a tendency to accumulate in perinuclear areas. These observations provide evidence that GAP‐43 is not restricted to neuronal cells. The close spatial association with cyto‐skeletal constituents, as observed in astrocytes. suggests a role for this protein in the control of cell shape, motility and adhesion processes. Copyright © 1995, Wiley Blackwell. All rights reserved
Expression and distribution of GAP-43 in human astrocytes in culture
Monacot S.;Rizzuto N.;
1995-01-01
Abstract
Expression and distribution of GAP‐43 in human astrocytes in culture By combining mRNA analysis and immunocytochemistry, we investigated the expression of the growth associated protein 43 (GAP‐43) in enriched populations of astrocytes, obtained from mixed cultures of human fetal brains. Total cellular RNA was extracted from cell pellets and reverse transcribed into cDNA; cDNA was subjected to PCR amplification using primers specific for GAP‐43 and PCR products were separated through polyacrylamide gels. Double immunofluorescence staining was performed on dissociated cell cultures using antibodies to glial fibrillary acidic protein (GFAP) and to GAP‐43. Results showed that both transcription and translation for GAP‐43 occur in cultured astrocytes. GAP‐43 immunoreacting material was detected in the cell processes and diffusely in the cytoplasm of GFAP‐positive astrocytes, during early stages of maintenance in vitro. In older cultures, GAP‐43 immunoreactivity persisted in a large percentage of cells, with a tendency to accumulate in perinuclear areas. These observations provide evidence that GAP‐43 is not restricted to neuronal cells. The close spatial association with cyto‐skeletal constituents, as observed in astrocytes. suggests a role for this protein in the control of cell shape, motility and adhesion processes. Copyright © 1995, Wiley Blackwell. All rights reserved
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