Brain endogenous insulin effects on neurite growth within fetal rat neuron cell cultures

We have previously described insulin to be synthesized "de novo" within the fetal rat brain and that brain endogenous insulin [I(n)] promoted neurofilament distribution within fetal neurons. In this study, we investigated the role of I(n) in neuron axonal growth. Rat fetal brain stem cells from 16-day gestational age were cultured in an IFDM and treated with an insulin antibody. In addition, the cell cultures were also treated in defined medium with the addition of: 5 ng, 20 ng or 100 ng/ml of insulin or 100 ng/ml insulin-like growth factor 1 (IGF-1). The neuron cell cultures were studied at 1 and 3 days of incubation. The presence of preproinsulin mRNA and insulin immunoreaction confirmed the "de novo" synthesis of insulin by the fetal neuron cell cultures. Axonal growth was similar by day 1 of the study in all the media, but in insulin medium containing 100 ng/ml of insulin the axonal length was significantly longer. By day 3 of incubation I(n) promoted axonal growth. Treating the neurons with an insulin antibody confirmed these findings, with a significant decrease in axonal length (p<0.05). The treatment with different concentrations of exogenous insulin did not promote axonal growth beyond I(n) by day 3 of incubation. IGF-1 did not promote axonal growth by day 3 of incubation. In summary, I(n) may promote axonal growth during brain development.