In vitro differentiation of neuronal precursor cells from amphibian late gastrulae: morphological, immunocytochemical studies, biosynthesis, accumulation and uptake of neurotransmitters

Neuronal differentiation has been studied in dissociated cell cultures from early neurulae of Pleurodeles waltl and Ambystoma mexicanum . Cocultures were prepared from the neural primordium and underlying chordamesoderm. NP and NF cultures were prepared from isolated neural plate and neural folds, respectively. Neuronal precursors in NP and NF cultures had distinctive aggregation properties already evident after 1–2 days in culture. After 10–15 days, mature neurones and synapses were observed by electron microscopy in the three culture types. The expression of neurofilament polypeptides and tetanus-toxin-binding sites was also present in these cultures. A small percentage of neurones contained cytochemically detectable catecholamine. Many neurones took up tritiated dopamine with a high affinity. Quantitative measurement of [ 3 H]acetylcholine synthesis and storage from [ 3 H]choline were negative at the early neurula stage and in 5 to 15-day-old NF cultures, and remained low in 5 to 15-day-old NP cultures. Acetylcholine production in cocultures increased linearly with time and was always much higher than in NP cultures. These results suggest that, at the early neurula stage, some neuronal precursors have acquired the capacity to express a high degree of morphological and biochemical differentiation even in the absence of further chordamesoderm influence. However, the chordamesodermal cells in the cultures increased acetylcholine synthesis.