Tryptophan hydroxylase mRNAs analysis by RT-PCR: preliminary report on the effect of noradrenaline in the neonatal rat pineal gland.
The levels of mRNAs coding for tryptophan hydroxylase (TPOH), the first enzyme in melatonin synthesis, have been investigated by quantitative reverse transcription of RNA, followed by polymerase chain reaction (RT-PCR), after stimulation of neonatal pineal organ cultures with Noradrenaline (NA). TPOH mRNAs were specifically amplified from various adult tissues, namely the pineal gland, raphe, retina, and kidney, but not the lung. PCR signals for TPOH were detected in the neonatal pineal gland in the absence of stimulation. Stimulation of neonatal pineal organ culture with 0.1 microM NA resulted in a significant increase (x2.5) in expression of TPOH mRNAs, whereas higher doses (1 and 10 microM) had no effect. All concentrations of NA enhanced melatonin secretion. Our results suggest that the level of TPOH mRNAs can be controlled by NA and that this effect might be implicated in the gene level regulation of the daily enzyme rhythm in the rat pineal gland.
Besançon
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INSERM U433, Laboratoire de Neurobiologie expérimentale et Physiopathologie, Faculté de Médecine R.T.H. Laënnec, Lyon, France. besancon@rockefeller1.univ-lyon1.fr
Reboul
A
A
Claustrat
B
B
Jouvet
A
A
Belin
M F
MF
Fèvre-Montange
M
M
D000818:Animals; D000831:Animals, Newborn; D017931:DNA Primers; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D008297:Male; D009638:Norepinephrine / Q000494:pharmacology*; D010870:Pineal Gland / Q000187:drug effects / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013566:Sympathomimetics / Q000494:pharmacology*; D014365:Tryptophan Hydroxylase / Q000235:genetics*
D017931:DNA Primers; D012333:RNA, Messenger; D013566:Sympathomimetics; D014365:Tryptophan Hydroxylase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D009638:Norepinephrine