Age-dependent differences in the effects of GDNF and NT-3 on the development of neurons and glia from neural crest-derived precursors immunoselected from the fetal rat gut: Expression of GFR alpha-1 in vitro and in vivo

No enteric neurons or glia develop in the gut below the rostral foregut in mice lacking glial cell line-derived neurotrophic factor (GDNF) or Ret. We analyzed the nature and age dependence of the effects of GDNF and, for comp arison, those of NT-3, on the in vitro development of the precursors of ent eric neurons and glia. Positive and negative immunoselection with antibodie s to p75(NTR) were used to isolate crest-derived and crest-depleted populat ions of cells from the fetal rat bowel at E12, 14, and 16. Cells were typed immunocytochemically. GDNF stimulated the proliferation of nestin-expressi ng precursor cells isolated at E12, but not at E14-16. GDNF promoted the de velopment of peripherin-expressing neurons (E12 much greater than E14-16) a nd expression of TrkC. GDNF inhibited expression of S-100-expressing glia a t E14-16. NT-3 did not affect cells isolated at E12, never stimulated precu rsors to proliferate, and promoted glial as well as neuronal development at E14-16. GFR alpha-1 was expressed both by crest- and non-crest-derived cel ls, although only crest-derived cells anchored GFR alpha-1 and GFR alpha-2 (GFR alpha-1 much greater than GFR alpha-2). GDNF increased the number of n eurons anchoring GFR alpha-1. GFR alpha-1 is immunocytochemically detectabl e in neurons of the E13 intestine and persists in adult neurons of both ple xuses. We suggest that GDNF stimulates the proliferation of an early (E12) NT-3-insensitive precursor common to enteric neurons and glia; by E14, this common precursor is replaced by specified NT-3-responsive neuronal and gli al progenitors. GDNF exerts a neurotrophic, but not a mitogenic, effect on the neuronal progenitor. The glial progenitor is not maintained by GDNF. (C ) 1998 Academic Press.