ISSN:
1432-0533
Schlagwort(e):
Neural cell surface antigens
;
Neural differentiation
;
Mouse teratoma
;
Radioimmune assay
;
Immunoperoxidase
Quelle:
Springer Online Journal Archives 1860-2000
Thema:
Medizin
Notizen:
Summary A rabbit antiserum against mouse neonatal brain cell surface membranes labeled by immunoperoxidase (PAP) the cells of the central and peripheral nervous systems of adult and neonatal mice and their processes, as well as the differentiating neuroepithelial cells of three OTT-6050 mouse teratoma-derived tumors. Indirect immunofluorescence on living 14-day-old monolayer cultures of neonatal mouse brain demonstrated reaction of the immune serum with external surface membrane antigens of neuroblasts and of primitive and mature glial cells. Radioimmune assays (RIA) showed almost complete loss of antiserum binding to neonatal mouse brain plasma membranes after absorption with adult or neonatal mouse brain membranes, and no loss of binding after absorption by liver, spleen, kidney, and heart membranes. Cross-reactivity of the immune serum to several non-neural cell types was demonstrated by immunoperoxidase on sperm and sperm-precursors, on moderate numbers of epithelial cells in the medulla of adult mouse thymus, and, in the neonate, on a range of mesenchymal cells. This cross-reactivity was reflected in the RIA by a moderate reduction of immune serum binding to neonatal mouse brain plasma membranes after absorption with testis pellets and with thymus membranes. PAP staining showed loss of crossreactivity after testis or thymus absorption, without climination of neural cell recognition. Absorption with adult or neonatal mouse brain eliminated cross-reactivity. In the teratoma-derived tumors, absorption of the antiserum with testis or thymus eliminated or markedly reduced the PAP staining of primitive neuroepithelial cells, and only moderately reduced, but did not remove, that of neural cells in the mature neuropil. Among the proteins of neonatal mouse brain plasma membranes separated by polyacrylamide gel electrophoresis, there were six distinct bands indicating major proteins ranging from 26,000–54,000 daltons. Autoradiography of the antigen-antibody complexes with125I protein A on the same gels demonstrated three discrete bands of activity at 10,000–12,000, 76,000, and 97,000 daltons, and one greater than 130,000 daltons, suggesting that the immune serum recognizes only minor protein components of the mouse brain plasma membranes. The application of the PAP method to the recognition of neural cell surface antigens considerably enhances the potential of this antiserum as a tool for the early identification of primitive neural cells in the experimental mouse teratoma.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1007/BF00690638
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