Characteristics of fish glial cells in culture: Possible implications as to their lineage

Abstract

Regeneration of injured central nervous system axons is largely dependent on the response of the associated nonneuronal glial cells to injury. Glial cells of the mammalian central nervous system, unlike those of fish, are apparently not conducive to axonal regeneration. While the lineage of rat glial cells is well characterized and its role in the support or inhibition of regenerative growth is beginning to be understood, little is known about fish glial cells. Accordingly, glial cells in cultures of adult goldfish brain and of newly hatched goldfish larvae were studied in an attempt to establish their lineage. The cells were identified by means of indirect immunofluorescence, using antibodies against fish astrocytes and oligodendrocytes. The cell count in the cultures increased from a small number of cells at 24 h after plating to a large number of both astrocytes and oligodendrocytes after 1 week in culture. Both of these cell types had originated from proliferating cells, as shown by their uptake of tritiated thymidine and by the inhibition of cell proliferation by 5-fluoro-2'-deoxyuridine. Both astrocytes, i.e., glial fibrillary acidic protein-positive cells, and oligodendrocytes, i.e., 6D2-positive cells, were positively labeled also by A2B5 antibodies, which are known to label progenitors of type-2 astrocytes and oligodendrocytes in the rat optic nerve. The results suggest that A2B5 positive progenitor cells in the goldfish central nervous system, as in the rat optic nerve, might be a common progenitor of astrocytes and oligodendrocytes.

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

“…Brain-derived primary cultures and cell lines have been established for several other fish species including tilapia [19], another African cichlid [41], goldfish [42], rainbow trout [43]–[45], barramundi [46], sea bass [47] and pompano [48]. We observed that a fibroblast-like phenotype prevailed after primary cells were kept in culture conditions for extended periods of time and overcame crisis.…”

Vimentin immunoreactive glial cells in the fish optic nerve: Implications for regeneration

“…Cultures of goldfish brain and indirect immunofluorescence of the cultured cells were carried out as described recently (Sivron et al, , 1992. Goldfish Longitudinal section (10pm) from an optic tract stained with FVPl and its phase, respectively.…”

The isolation and culture of microglia-like cells from the goldfish brain

“…Finally, we differ from other workers in our choice of substrates. Nearly all previous studies used a polycation (poly-l-lysine, -ornithine, or poly-d-lysine) coated substrate although two groups used polycation/laminin coated dishes to grow their cells (Bastmeyer et al, 1994;Sivron et al, 1992). In our hands, we found that microglia could only be isolated on completely untreated, bacterial-grade plastic.…”

“…While there have been no previous studies of teleost microglia in vitro, there are numerous reports describing isolation protocols for teleost macroglia-astrocytes and oligodendrocytes (Bastmeyer et al, 1989;Clasen et al, 1995;Gould et al, 1995;Jeserich and Rauen, 1990;Sivron et al, 1992;Tocher and Wilson, 1990). The procedure that we describe here differs in most respects from these earlier protocols and it is interesting that, with the exception of Sivron et al (1992), none of these papers reported the presence of microglial cells in their cultures.…”

“…The procedure that we describe here differs in most respects from these earlier protocols and it is interesting that, with the exception of Sivron et al (1992), none of these papers reported the presence of microglial cells in their cultures. Most workers used L15 medium (Anderson et al, 1987;Bastmeyer et al, 1990;Gould et al, 1995;Sivron et al, 1992) although both Ham's F12 (Bastmeyer et al, 1993;Hoppe et al, 1990) and Dulbecco's modification of Eagle's medium (DMEM) (Jeserich and Rauen, 1990;Tocher and Wilson, 1990) have been used as well. In our cultures, we found that cells did not grow well in L15 and that we could not effectively buffer DMEM against pH changes in air.…”