Role of basic FGF and oxygen in control of proliferation, survival, and neuronal differentiation in carotid body chromaffin cells

Abstract

Crest-derived glomus cells of the carotid body (CB) are O2-sensitive chemoreceptors, which resemble sympathoadrenal (SA) chromaffin cells. In this study, we tested whether perinatal rat glomus cells are sensitive to basic fibroblast growth factor (bFGF) in vitro and whether their sensitivity is regulated by oxygen. In chemically defined medium, bFGF (1-100 ng/ml) caused a significant, dose-dependent increase in the number of surviving tyrosine hydroxylase-positive (TH+) glomus cells in embryonic (E17-E19) CB cultures, following a 48-hr exposure. Though basic FGF (10 ng/ml) appeared mitogenic for these cells, based on stimulation of bromodeoxyuridine (BrdU) uptake, it supported survival of only ~60% of the initial TH+ population, suggesting that significant cell death was occurring. This apparent cell loss in E17 cultures could be largely prevented by combined treatment with bFGF and low oxygen (6% O2). In contrast, in early postnatal (P1) cultures, glomus cell number was relatively unchanged over 48 hr under control conditions or in presence of mitogenic activity from either bFGF or low oxygen. However, combined treatment with both bFGF and low oxygen stimulated proliferation of P1 glomus cells such that by 48 hr the TH+ population had increased to ~1.5x the initial density. Basic FGF (10 ng/ml) also stimulated neurite outgrowth and neurofilament expression in E18-E19, but not P1-P3, glomus cells. In contrast to bFGF, treatment with nerve growth factor was ineffective. Taken together, these results suggest that bFGF and low oxygen are mitogens for perinatal CB chromaffin cells and interact cooperatively as survival factors. It is plausible that these mechanisms may operate to regulate chemoreceptor cell density, during the animal's transition from in utero (hypoxic) to ex utero (normoxic) life.