Characterization of a novel primary culture system of adult zebrafish brainstem cells

Abstract

Adult zebrafish (Danio rerio) have a remarkable ability to restore function after an injury to the brain or spinal cord. The molecular and cellular mechanisms underlying this phenomenon are not fully understood. To enable investigation of these mechanisms we have developed an in vitro model system from the adult zebrafish brainstem, which can be maintained under serum-containing and serum-free conditions. While cultures are predominantly neuronal, they also contain glia and stem progenitor cells. Various stages of cellular differentiation are observed among both neuronal and non-neuronal populations. Quantitative morphological results revealed typical cellular growth over a two-week period. We argue that our novel brainstem culture model offers a powerful tool for the studies of axonal growth, neurogenesis, and regeneration in the adult zebrafish central nervous system.

Keywords: 5-bromo-2′-deoxyuridine; ANOVA; Brainstem; BrdU; CNS; DNase; Danio rerio; EDTA; FACS; FBS; GFAP; Glia; HuC/D; MAb; Neuron; PAb; PBS; PCNA; Regeneration; SEM; Stem cell; Tph2; analysis of variance; central nervous system; days in vitro; deoxyribonuclease; div; ethylenediaminetetraacetic acid; fetal bovine serum; fluorescence-activated cell sorting; glial fibrillary acidic protein; human neuronal protein C/D; monoclonal antibody; phosphate-buffered saline; polyclonal antibody; proliferating cell nuclear antigen; standard error of the mean; tryptophan hydroxylase 2.