Author/Editor		Miš, Katarina; Marš, Tomaž; Jevšek, Marko; Brank, Martina; Zajc-Kreft, Katarina; Grubič, Zoran
Title		Localization of mRNAs encoding acetylcholinesterase and butyrylcholinesterase in the rat spinal cord by nonradioactive in situ hybdrization
Type		članek
Source		J Histochem Cytochem
Vol. and No.		Letnik 51, št. 12
Publication year		2003
Volume		str. 1633-44
Language		eng
Abstract		In spite of intensive investigations, the roles of acetylcholinesterase (AChE; EC 3.1.1.7) and butyrylcholinesterase (BuChE; EC 3.1.1.8) in the central nervous system (CNS) remain unclear. A role recently proposed for BuChE as an explanation for survival of AChE knockout mice is compensation for AChE activity if it becomes insufficient. Neuronal contribution of both enzymes to the cholinesterase pool in the neuromuscular junction has also been suggested. These proposals imply that BuChE expression follows that of AChE and that, in addition to AChE, BuChE is also expressed in a-motor neurons. However, these assumptions have not yet been properly tested. Histochemical approaches to these problems have been hampered by a number of problems that prevent unambiguous interpretation of results. In situ hybridization (ISH) of mRNAs encoding AChE and BuChE, which is the state-of-the-art approach, has not yet been done. Here we describe rapid nonradioadive ISH for the localization of mRNAs encoding AChE and BuChE. Various probes and experimental conditions had been tested to obtain reliable localization. In combination with RT-PCR, ISH revealed that, in rat spinal cord, cells expressing AChE mRNA also express BuChE mRNA but in smaller quantities. a-Motor neurons had the highest levels of both mRNAs. Virtual absence of transcripts encoding AChE and BuChE in glia might reflect a discrepancy between mRNA and enzyme levels previously reported for cholinesterases.
Descriptors		SPINAL CORD RNA, MESSENGER ACETYLCHOLINESTERASE BUTYRYLCHOLINESTERASE RATS, WISTAR POLYMERASE CHAIN REACTION RNA PROBES IN SITU HYBRIDIZATION IMMUNOHISTOCHEMISTRY FLUORESCENT DYES CELL COUNT