Identification of serotonin transporter mRNA in rat platelets.
Total RNA isolated from rat platelets by guanidinium-acid-phenol extraction, and mRNA for the serotonin (5-hydroxytryptamine) transporter (5HTt) was identified. From a typical starting sample of 20 mL of rat blood (approximately 9 x 10(9) platelets), 14 to 17 micrograms of total platelet RNA was obtained. Northern blot analysis, using 32P-labeled 5HTt cDNA as a probe, identified approximately 3.3 kb long 5HTt mRNA. After rehybridization with cyclophilin cDNA, approximately 1 kb long mRNA for cyclophilin, which could serve as a reference for 5HTt mRNA quantification, was also identified. Densitometric analysis demonstrated clearly measurable signals for both mRNAs. The possibility of quantification of rat platelet 5HTt mRNA should enable parallel studies on 5HTt gene expression in platelets and brain of the same animal, and the evaluation of the platelet model at the molecular genetic level.
Hranilović
D
D
Department of Organic Chemistry and Biochemistry, Ruder Bosković Institute, Zagreb, Croatia.
Lesch
K P
KP
Ugarković
D
D
Cicin-Sain
L
L
Jernej
B
B
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000097:blood / Q000235:genetics*; D005260:Female; D008297:Male; D008562:Membrane Glycoproteins / Q000097:blood / Q000235:genetics*; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger / Q000097:blood*; D051381:Rats; D017208:Rats, Wistar; D050486:Serotonin Plasma Membrane Transport Proteins
D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D050486:Serotonin Plasma Membrane Transport Proteins; C493232:Slc6a4 protein, rat