Localization of substance P mRNA in cholinergic cells of the rat laterodorsal tegmental nucleus: in situ hybridization histochemistry and immunocytochemistry.
1. In situ hybridization histochemical techniques in combination with immunocytochemistry and acetylcholinesterase (AChE) histochemistry were used to study the colocalization of messenger RNA (mRNA) encoding the neuropeptide substance P (SP) in cholinergic cells of the laterodorsal tegmental nucleus (LDT) of the rat pontine brain stem. 2. Alternate serial sections were hybridized with a 48-base, 35S-labeled synthetic oligonucleotide probe encoding SP using in situ hybridization histochemistry and processed either histochemically for AChE or immunocytochemically for choline acetyltransferase (ChAT). 3. In addition, serial section analysis was used to demonstrate the correlation between SP and SP mRNA in the same cells of the LDT. 4. These studies reveal that the cholinergic neurons of the LDT synthesize SP.
Laboratory of Clinical Science, National Institute of Mental Health, Bethesda, Maryland 20892.
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Sutin
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Jacobowitz
D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D002799:Cholinergic Fibers / Q000378:metabolism*; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D008297:Male; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000378:metabolism; D011149:Pons / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013373:Substance P / Q000235:genetics* / Q000378:metabolism
D009841:Oligonucleotides; D012333:RNA, Messenger; D013373:Substance P; D002795:Choline O-Acetyltransferase; D000110:Acetylcholinesterase