Early expression of a novel radial glia antigen in the chick embryo

Abstract

Monoclonal antibody 3CB2 recognizes an antigen expressed in discrete cell types derived from ectoderm and mesoderm. Biochemical and immunohistochemical studies indicate that the antigen recognized by the antibody is a 55 kDa cytoplasmic protein that may be an intermediate filament associated protein (IFAP). Developmental studies show that 3CB2 antigen is intensely expressed very early in the chick embryo, in the neural tube, myotomes, and in mesenchymal cells of visceral arches and the presumptive facial area. As development proceeds, antigen expression becomes restricted to astrocytes and radial glia cells throughout the brain. A detailed immunohistochemical study of the developing chick retina shows that the expression of 3CB2 antigen at embryonic day 8 (E8) is restricted to Müller cells, with the pattern of expression closely related to their degree of differentiation. We show, by immunocytochemistry labeling of entire Müller cells dissociated from retinas of E16-E20, that 3CB2 monoclonal is labeling the whole cell. 3CB2 selectively labels Müller cells in the rat and chameleon, but not their retinal horizontal cell axons. 3CB2 monoclonal is a very sensitive marker for early differentiating Müller cells. Our results provide evidence that 3CB2 antigen is a cytoskeletal component which may be involved in the morphogenesis of these cells, and also perhaps in the outgrowth of some axons.

Human ES cell-derived neural rosettes reveal a functionally distinct early neural stem cell stage

“…Most rosette cells expressed Pax6, while a subset of these cells coexpressed 3CB2 (Fig. 1H), a marker of radial glia (Prada et al 1995;Conti et al 2005). Immunocytochemical analyses confirmed the neural stem cell, neuroepithelial, and anterior CNS character of hESC-derived neural rosettes ( Fig.…”

Stable expression of intracellular Notch suppresses v-Src-induced transformation in avian neural cells

“…Moreover, we could not detect expression of neuronal markers (data not shown). The lack of Pax6 and Chx10 expression, combined with the presence of vimentin is characteristically found in NR glial cells in ovo (Alvarez-Buylla et al, 1987;Prada et al, 1995;Marquardt and Gruss, 2002). Reactivation of v-Src, upon transferring the cultures to 371C, did not influence the pattern of markers expression ( Figure 3B, j, l, n, p).…”

Implication of OTX2 in Pigment Epithelium Determination and Neural Retina Differentiation

“…mAb 40.2D6, directed against islet-1, used to identify mature RGCs (Austin et al, 1995), was obtained from the Developmental Studies Hybridoma Bank and was used in a 1:1000 dilution of ascitic fluid. The 3CB2 mAb was used, in a 1:500 dilution of ascitic fluid, to specifically identify Müller cells (Prada et al, 1995). The mAb against BrdU was purchased from Boehringer Mannheim and used at a 1:200 dilution.…”

“…If OTX2 expression was a common feature in the differentiation of retina neuroepithelial cells, then it should be anticipated that Müller cells also express OTX2. 3CB2 is a mAb directed against an intermediate filament-related antigen, which selectively labels Müller cells as they leave the cell cycle at E7 (Prada et al, 1995). Thus, E7 chick retinas were double stained with anti-OTX2 and -3CB2.…”