Localization of insulin-like immunoreactivity in the neurons from primary cultures of rat brain

Brains from 1-day-old rats were dissociated with trypsin and the cells were maintained in culture for 3-4 weeks. These primary cultures contained insulin-like immunoreactivity in the limited populations of neurons. Typically, the fluorescent staining pattern observed in the soma was homogeneous and varicosity-like structures were observed on the neurites of the majority of insulin-like immunoreactive neurons. Serum deprivation of brain cell cultures did not reduce immunoreactivity, whereas cycloheximide caused approx. 80% decrease in the number of insulin-like immunoreactive neurons. Incubation of these cultures with [3H]valine resulted in the incorporation of radioactivity into immunoprecipitable insulin. These results suggest that insulin-like immunoreactivity present in the Central Nervous System (CNS) neurons may be synthesized by brain cell cultures.