Neurite promoting activity of insulin, insulin-like growth factor I and nerve growth factor on spinal motoneurons is astrocyte dependent.

Mouse motoneurons were isolated from dissociated E15 mouse spinal cord and grown on polyornithine-coated round coverslips in a growth medium (DMEM/F12) supplemented with progesterone, trans-ferrin, selenium, horse serum and muscle extract. Astrocytes from newborn mouse neopallium were grown on rectangular coverslips. The motoneuron neurite growth was determined at day 8 of culture by counting, using the light microscope, the intersections produced by neurites radiating from the perikaryon placed centrally in a graticule eyepiece of concentric circles. The mean intersections for cultures without addition of astrocytes, insulin, insulin-like growth factor I (IGF-I) or nerve growth factor (NGF) was 12.6 +/- 0.8. When astrocytes on a separate coverslip were introduced from day 1, there was a small increase in neurite growth (16.3 +/- 0.9). The neurite growth was further increased significantly with the addition of insulin (27.3 +/- 1.4), IGF-I (31.5 +/- 1.4) or NGF (21.8 +/- 1.1) to cultures with astrocytes. Insulin, IGF-I or NGF in the absence of astrocytes did not greatly increase the neurite growth. We conclude that insulin, IGF-I and NGF promote neurite growth through some interactions with astrocytes.