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Problems encountered when immunocytochemistry is used for quantitative glial cell identification in autoradiographic studies of cell proliferation in the brain of the unlesioned adult mouse (1994)

Korr, Hubert ; Horsmann, Cornelia ; Schürmann, Martin ; [et al.]

Springer

Cell & tissue research 278 (1994), S. 85-95

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ISSN: 1432-0878

Keywords: Key words: Autoradiography ; Immunocytochemistry ; Astrocytes ; Oligodendrocytes ; Cell proliferation ; Mouse (Han: NMRI)

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Abstract. We have used sections of adult mouse brain to determine whether antibodies specific for oligodendroglia (anti-carbonic anhydrase II, CA II; anti-galactocerebroside, GC; anti-myelin basic protein, MBP) and astroglia (anti-glial fibrillary acidic protein, GFAP; anti-S 100 protein) are suitable for quantitative studies of the proliferation and subsequent differentiation of these cells. Unlesioned adult mice received a single injection of 3H-thymidine (TdR) and were killed between 1 h and 70 days later. Quantitative evaluations of autoradiographs of 2-μm-thick serial sections stained immunocytochemically with the antibodies mentioned above or with Richardson's method for histological control led to the following conclusions. Anti-GC and anti-MBP stained only the oligodendrocytic processes and, thus, cannot be used in well-myelinated brain areas. Anti-CA II stained only a portion of the differentiated oligodendrocytes, but no proliferating cells. Anti-S 100 protein recognized all the astrocytes, but also many (interfascicular) oligodendrocytes. Anti-GFAP stained only a few astrocytes in the unlesioned mouse; all astrocytes may become GFAP-immunopositive only after wounding the brain. Thus, in contrast to in vitro studies, immunocytochemical studies with these antibodies on sections of adult animals cannot be recommended for the quantitative analysis of cell proliferation. In addition, our results show that differentiated glial cells proliferate in adult mice. Astro- and oligodendrocytes divide with the same cell cycle parameters and mode of proliferation up to about 1 month after 3H-TdR injection. In contrast to oligodendrocytes, some astrocytes might re-enter the cycle after a few weeks of quiescence.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00305780

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Long-term induction of an aldose reductase protein by basic fibroblast growth factor in rat astrocytes in vitro (1995)

Laeng, Pascal ; Bouillon, Philippe ; Taupenot, Laurent ; [et al.]

Weinheim : Wiley-Blackwell

Electrophoresis 16 (1995), S. 1240-1250

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ISSN: 0173-0835

Keywords: Basic fibroblast growth factor ; Astrocytes ; Cell maturation ; Aldose reductase ; Two-dimensional polyacrylamide gel electrophoresis ; Protein microsequencing ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Chemistry and Pharmacology

Notes: Basic fibroblast growth factor (bFGF) is known to elicit various developmental-like effects on astrocytes in vitro, but these effects were studied mainly over short-term periods. In this work we asked the question whether bFGF could induce long-term effects on rat astrocytes in culture. This factor was found to induce only a transient mitogenic effect lasting less than 48 h, even when the treatment was carried on for 4 days. By contrast, it induced long-term effects on the rate of synthesis of several proteins as seen by two-dimensional polyacrylamide gel electrophoresis after labeling the cells with [35S]methionine. The most upregulated protein was extracted from preparative gels of soluble extracts of cultured bFGF-treated astrocytes and of normal brain. It was characterized by internal amino acid microsequencing. Two tryptic digest peptides had N-terminal sequences similar to rat lens aldose reductase. This protein was also expressed in oligodendroglial and neuronal cells in culture, but it was not upregulated by bFGF. Aldose reductase is thought to be involved in a minor pathway of glucose metabolism and in diabetic complications. Its longterm regulation by bFGF will possibly help in the understanding of its actual physiological role.

Additional Material: 10 Ill.