Development of cholinergic terminals around rat spinal motor neurons and their potential relationship to developmental cell death.
Neuron death seems to be regulated mainly by postsynaptic target cells. In chicks, nicotinic antagonists such as alpha-bungarotoxin (alphaBT) can prevent normal cell death of somatic motor neurons (SMNs). For this effect, however, alphaBT could be acting at peripheral neuromuscular junctions and/or central cholinergic synapses. To investigate this issue, we first studied the development of cholinergic terminals in the rat spinal cord by using vesicular acetylcholine transporter immunocytochemistry. Labeled terminals were seen in the ventral horn as early as embryonic day 15 (E15), the beginning of the cell death period. Thus, central cholinergic synapses form at the correct time and place to be able to influence SMN death. We next added alphaBT to organotypic, spinal slice cultures made at E15. After 5 days in vitro, the number of SMNs in treated cultures was substantially greater than in control cultures, indicating that alphaBT can reduce SMN cell death in rats as it does in chicks. Moreover, peripheral target removal led to extensive loss of SMNs, and such a loss occurred even in the presence of alphaBT, indicating the necessity of peripheral target for the alphaBT effect. Finally, to determine whether central cholinergic terminals also may be involved in SMN death, we delayed the alphaBT treatment until after central cholinergic terminals had disappeared from the slice cultures. The increased number of surviving SMNs, even in the absence of central terminals, argued that alphaBT acts at peripheral, not central, cholinergic synapses to rescue SMNs from developmental cell death.
Wetts
R
R
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010-3011, USA. rwetts@uci.edu
Vaughn
J E
JE
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D018680:Cholinergic Antagonists / Q000494:pharmacology; D002799:Cholinergic Fibers / Q000502:physiology*; D004622:Embryo, Mammalian / Q000502:physiology; D005314:Embryonic and Fetal Development; D005260:Female; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D026901:Membrane Transport Proteins; D009046:Motor Neurons / Q000502:physiology*; D009411:Nerve Endings / Q000254:growth & development* / Q000378:metabolism; D051381:Rats / Q000196:embryology* / Q000254:growth & development; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D050494:Vesicular Acetylcholine Transport Proteins; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D018680:Cholinergic Antagonists; D026901:Membrane Transport Proteins; C493461:Slc18a3 protein, rat; D050494:Vesicular Acetylcholine Transport Proteins; D033921:Vesicular Transport Proteins