Partial cloning and expression of mRNA coding choline acetyltransferase in the spinal cord of the goldfish, Carassius auratus.
Choline acetyltransferase (ChAT, EC 2.3.1.6) synthesizes a neurotransmitter, acetylcholine in cholinergic neurons. ChAT is considered to be the most specific marker for cholinergic neurons. To obtain a better marker of the neurons, as the first step, we isolated a partial ChAT cDNA from the goldfish (Carassius auratus) brain by RT-PCR methods. The partial cDNA of the goldfish ChAT was composed of 718 nucleotides. The amino acid sequence of the goldfish ChAT is approximately 70% identical to those of mammalian and chicken ChAT. Northern blot analysis demonstrated that ChAT mRNA was expressed in the brain and the spinal cord of the goldfish, and much abundant in the spinal cord. In the spinal cord of the goldfish, ChAT-positive neurons were detected mainly in the ventral horn by in situ hybridization. In addition, fluorescence in situ hybridization combined with a retrograde labeling by using True Blue demonstrated ChAT mRNA positive neurons were exactly motoneurons. In the cord, putative presynaptic sympathetic neurons were also labeled.
Moon
Soon-Ju
SJ
Laboratory of Fish Physiology, Graduate School of Biosphere Sciences, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
Fujikawa
Yukichi
Y
Nishihara
Takahiro
T
Kono
Sayuri
S
Kozono
Keina
K
Ikenaga
Takanori
T
Esaka
Muneharu
M
Iijima
Noriaki
N
Nagamatsu
Yasunori
Y
Yoshida
Masayuki
M
Uematsu
Kazumasa
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002795:Choline O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D006054:Goldfish / Q000235:genetics / Q000378:metabolism*; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009474:Neurons / Q000201:enzymology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013116:Spinal Cord / Q000201:enzymology*
D012333:RNA, Messenger; D002795:Choline O-Acetyltransferase