Trophic support of cultured spiral ganglion neurons by depolarization exceeds and is additive with that by neurotrophins or cAMP and requires elevation of [Ca2+]i within a set range.
Spiral ganglion neurons (SGNs) require both pre- and postsynaptic contacts to maintain viability. BDNF, NT-3, chlorphenylthio-cAMP, and depolarization (veratridine or elevated [K+]o) all promote survival of SGNs in vitro, depolarization being the most effective. Combining different trophic stimuli increases survival in an additive manner. Neurotrophins and depolarization maintain comparable soma size and neurite extension, but SGNs are shrunken in cAMP. Elevated [K+]o has a biphasic effect on SGN survival; survival improves as [K+]o is raised to 30 mM (30K) and falls as [K+]o is further increased; SGN survival in 80 mM [K+]o (80K) is poor relative to survival in 30K. These responses to elevated [K+]o are potentiated by an L-type channel agonist, whereas L-type Ca2+ channel blockers antagonize the trophic effect of depolarization. Four hours after depolarization, steady-state [Ca2+]i is elevated in SGNs in 30K and further elevated in SGNs in 80K. At 22 hr after depolarization, by which time death of neurons in 80K has begun, elevated [Ca2+]i levels in surviving neurons in 80K are not higher than those in neurons in 30K ( approximately 150-450 nM), suggesting that neurons with high [Ca2+]i are preferentially lost. Veratridine causes oscillatory increases in [Ca2+]i to 250-350 nM. Thus, [Ca2+]i is predictive of cell survival; [Ca2+]i elevated to 100-500 nM in a sustained or oscillatory manner permits SGN survival independent of exogenous neurotrophic factors. Higher [Ca2+]i is associated with cell death.
Hegarty
J L
JL
Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242, USA.
Kay
A R
AR
Green
S H
SH
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology*; D009414:Nerve Growth Factors / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013136:Spiral Ganglion / Q000187:drug effects*
D019208:Brain-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D000242:Cyclic AMP; D002118:Calcium