Expression of substance P and preprotachykinin mRNA by primary sensory neurons in culture: regulation by factors present in peripheral and central target tissues.
Primary sensory neurons display various neuronal phenotypes which may be influenced by factors present in central or peripheral targets. In the case of DRG cells expressing substance P (SP), the influence of peripheral or central targets was tested on the neuronal expression of this neuropeptide. DRG cells were cultured from chick embryo at E6 or E10 (before or after establishment of functional connections with targets). Preprotachykinin mRNA was visualized in DRG cell cultures by either Northern blot or in situ hybridization using an antisense labeled riboprobe, while the neuropeptide SP was detected by immunostaining with a monoclonal antibody. In DRG cell cultures from E10, only 60% of neurons expressed SP. In contrast, DRG cell cultures performed at E6 showed a significant hybridization signal and SP-like immunoreactivity in virtually all the neurons (98%). The addition of extracts from muscle, skin, brain or spinal cord to DRG cells cultured at E6 reduced by 20% the percentage of neurons which express preprotachykinin mRNA and SP-like immunoreactivity. Our results indicate that factors issued from targets inhibit SP-expression by a subset of primary sensory neurons and act on the transcriptional control of preprotachykinin gene.
Barakat-Walter
I
I
Institut d'Histologie et d'Embryologie, Faculté de Médicine, Université de Lausanne, Switzerland.
Affolter
H U
HU
Droz
B
B
D000818:Animals; D001345:Autoradiography; D015152:Blotting, Northern; D001921:Brain / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D004261:DNA Replication; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression; D005786:Gene Expression Regulation; D009132:Muscles / Q000502:physiology; D009475:Neurons, Afferent / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010761:Phosphorus Radioisotopes; D011498:Protein Precursors / Q000235:genetics*; D012313:RNA / Q000302:isolation & purification; D015347:RNA Probes; D012333:RNA, Messenger / Q000235:genetics*; D012879:Skin Physiological Phenomena; D013116:Spinal Cord / Q000502:physiology; D013373:Substance P / Q000235:genetics*; D013462:Sulfur Radioisotopes; D015320:Tachykinins / Q000235:genetics*; D014020:Tissue Extracts / Q000494:pharmacology
D010761:Phosphorus Radioisotopes; D011498:Protein Precursors; D015347:RNA Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D015320:Tachykinins; D014020:Tissue Extracts; C044206:preprotachykinin; D013373:Substance P; D012313:RNA