Peripheral and central target requirements for survival of embryonic rat dorsal root ganglion neurons in slice cultures.
Developmental cell death in the nervous system usually is controlled by the availability of target-derived trophic factors. It is well established that dorsal root ganglia (DRG) neurons require the presence of their peripheral target for survival, but because of their central projections, it is possible that the spinal cord also may be required. Before examining this possibility in rat embryos, we first used terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) to determine that thoracic DRG cell death occurred from embryonic day 15 (E15) to E18. To determine the target requirements of DRG neurons, we used organotypic slice cultures of E15 thoracic trunk segments. After peripheral target removal, essentially all DRG neurons disappeared within 5 d. In contrast, after removal of the spinal cord, approximately half of the DRG neurons survived for at least 8 d. Hence, some E15 DRG neurons could survive without the spinal cord. However, those DRG neurons that died after spinal cord ablation apparently required trophic factors from both central and peripheral targets, because the presence of only one of these tissues was not adequate by itself to support this cell group. Addition of neurotrophin-3 (NT-3) to the culture medium rescued some DRG neurons after CNS removal, suggesting a possible role for NT-3 in vivo. In other experiments, cultures were established from older (E16) embryos, and essentially all neurons survived after spinal cord ablation, even without added factors. These and other experiments indicated that approximately 65% of DRG neurons are transiently dependent on the CNS early in development.
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
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Wetts
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Vaughn
D000818:Animals; D002470:Cell Survival / Q000502:physiology; D053938:DNA Fragmentation; D004253:DNA Nucleotidylexotransferase; D003856:Deoxyuracil Nucleotides; D004622:Embryo, Mammalian / Q000166:cytology; D005314:Embryonic and Fetal Development / Q000502:physiology; D005727:Ganglia, Spinal / Q000166:cytology / Q000196:embryology*; D006651:Histocytochemistry; D009414:Nerve Growth Factors / Q000494:pharmacology; D009474:Neurons / Q000166:cytology*; D020933:Neurotrophin 3; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley
D003856:Deoxyuracil Nucleotides; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D004253:DNA Nucleotidylexotransferase