EXPRESSION OF MECHANO-SENSITIVE RECEPTOR GENES IN THE DETRUSOR OF THE RAT BLADDER

Abstract

B a c k g r o u n d . Autonomic mechanosensitivity of the muscle layer (detrusor) of the urinary bladder is an actual problem that still remains understudied. It is known that a number of mechanosensitive channels are probably expressed, but to date, only the leading role of the mechanosensitive channel TREK-1 has been established. The aim of the study was to check the expression of mechanoreceptor genes of smooth muscle tissue (KCNK2 (TREK-1), TRPV4, Piezo1, and Piezo2) in the bladder detrusor. M e t h o d s . Unique primers were selected for the detection of the specified channel genes, as well as the reference gene GAPDH, using the BLAST software. Validation was performed using one-step PCR with reverse transcription on the CFX96 amplifier with fluorescence detection of the Fam sample. A matrix RNA sample was isolated from the muscle and urothelial layers of the urinary bladder and dorsal root ganglia (DRG) of healthy Wistar rats using a commercial nucleic acid extraction column kit (Biocorp, Ukraine). R e s u l t s . In our samples, Piezo1 and TREK-1 channels were shown to be expressed in the urothelial and muscle layers of the bladder, whereas TRPV4 was only expressed in the urothelium. Piezo2 gene expression was not detected. C o n c l u s i o n s . The selected sequences of primers for TREK-1, Piezo1, TRPV4, and the "housekeeping" gene GAPDH are effective for studying the expression of these genes in rats. The synthesized sequences correspond to the expected fragment sizes. The genes of three mechanosensitive channels are expressed in the urinary bladder: TREK-1, Piezo1, and TRPV4.