Gene expression of catecholamine synthesizing enzymes in A5 cell group and modulation of tyrosine hydroxylase mRNA by immobilization stress.
OBJECTIVE
The A5 group of noradrenergic neurons plays a key role in autonomic mechanisms like cardiovascular regulation, nociception and respiration. The aim of this work was to detect the gene expression of catecholamine synthesizing enzymes in A5 brain nuclei.


METHODS
The gene expression of. tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine N-methyl-transferase (PNMT) in A5 brain nuclei was estimated. We also investigated various time intervals after the end of the single two-hour immobilization, as well as the effect of short-term repeated immobilization (120 min daily for 7 days) on tyrosine hydroxylase gene expression, the rate-limiting enzymes in catecholamines biosynthesis, in the A5 cell group. For all experiments, reverse transcription with subsequent polymerase chain reaction (RT-PCR) was used.


RESULTS
As expected, we detected a clear signal for TH and DBH mRNA but no signal for PNMT mRNA. Both, single and repeated immobilization stress exposure increased significantly the gene expression of TH in A5 area. Maximal elevation in TH mRNA levels occurred after single immobilization for two hours and subsequent decapitation 24 hours later.


CONCLUSIONS
In this study we detected for the first time the presence of DBH mRNA in micro dissected A5 cell group. We also showed how the gene expression of tyrosine hydroxylase changed with the function of time after the single immobilization exposure. Thus, TH mRNA in A5 cell group is modulated by immobilization stress in a time-dependent manner.
Micutkova
L
L
Institute of Experimental Endocrinology, Slovak Academy of Science, Vl rska 3, 833 06 Bratislava, Slovak Republic. ueenmicu@savba.sk
Kiss
A
A
Filipenko
M
M
Rychkova
N
N
Krizanova
O
O
Palkovits
M
M
Kvetnansky
R
R
D000818:Animals; D001921:Brain / Q000201:enzymology*; D004299:Dopamine beta-Hydroxylase / Q000235:genetics*; D015870:Gene Expression; D008297:Male; D009474:Neurons / Q000201:enzymology; D009638:Norepinephrine / Q000502:physiology; D010625:Phenylethanolamine N-Methyltransferase / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012149:Restraint, Physical; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013312:Stress, Physiological / Q000201:enzymology*; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*
D012333:RNA, Messenger; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase; D010625:Phenylethanolamine N-Methyltransferase; D009638:Norepinephrine