Evidence for tryptophan hydroxylase and hydroxy-indol-O-methyl-transferase mRNAs in human blood platelets.
Human blood platelets were tested for the presence of mRNAs coding for tryptophan hydroxylase (TPOH) and hydroxy-indol-o-methyl-transferase (HIOMT). Total RNA was extracted from platelets (12.9 +/- 3.3 mg RNA/100 ml blood, mean +/- SEM of 6 preparations) and cDNA synthesized by reverse transcription using random hexamers, oligo-dT or TPOH- or HIOMT-specific primers, designed to amplify a 254 bp fragment for TPOH and a 301 bp fragment for HIOMT. Positive controls were performed using RNA extracted from human normal or tumoral pineal glands. The PCR products were analyzed by gel electrophoresis, transferred to a nylon membrane and hybridized with a 32P-labeled internal probe. When random hexamers, oligo-dT or specific primers were used for reverse transcription, amplification products of the predicted sizes were detectable following electrophoresis in the case of pineal glands and following transfer and hybridization in the case of platelets. These results show TPOH and HIOMT mRNAs to be present in human blood and support the hypothesis that serotonin and melatonin may be synthesized in blood and, more particularly, in platelets.
Champier
J
J
INSERM U433, Faculté de Médecine Rene-Laénnec, Lyon, France. champier@laennec.univ-lyon1.fr
Claustrat
B
B
Besançon
R
R
Eymin
C
C
Killer
C
C
Jouvet
A
A
Chamba
G
G
Fèvre-Montange
M
M
D000121:Acetylserotonin O-Methyltransferase / Q000097:blood / Q000235:genetics*; D001792:Blood Platelets / Q000737:chemistry*; D015139:Blotting, Southern; D004587:Electrophoresis, Agar Gel; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000097:blood*; D014365:Tryptophan Hydroxylase / Q000097:blood / Q000235:genetics*
D012333:RNA, Messenger; D014365:Tryptophan Hydroxylase; D000121:Acetylserotonin O-Methyltransferase