Preferential growth of neurites from isolated fetal mouse dorsal root ganglia in relation to specific regions of co-cultured spinal cord explants

Abstract

Clusters of dorsal root ganglia (DRGs) from 13- to 14-day fetal mice were co-cultured with specific fragments of deafferented spinal cord (0.5-1 mm apart) on collagen-coated coverslips in Maximow slide chambers. Nerve growth factor (NGF) was added to the culture medium (1000 biological units/ml, at explantation) to ensure optimal survival and growth of a large fraction of the fetal DRG neurons. Sequential microscopic observations of the living cultures and cytologic studies after silver impregnation demonstrate that many neurites from isolated DRGs can invade dorsal (DC) regions of co-cultured spinal cord explants, whereas they are deflected from ventral cord (VC) tissue and its neuritic-glial outgrowth. Furthermore, some DRG neurites may become redirected towards distant DC target explants even after long circuitous detours around proximally arrayed VC explants. DRG neurites also show remarkably sharp projections to DC tissue and more complete avoidance of adjacent VC tissue when the DRG neurites approach a suitably arranged DC-VC-DC interface, e.g. forming de novo 'dorsal roots' at the end of a longitudinal strip of whole spinal cord. These experiments suggest that DC-VC boundaries may be particularly effective in guiding DRG neurites to specific regions of the CNS. The present studies of co-cultured fetal mouse DRG and spinal cord explants provide the first demonstration of preferential neuritic growth in vitro in relation to specific CNS target tissues.