[Primary study in rabbit penile corpus cavernosum smooth muscle cell culture and RyRs expression].
OBJECTIVE
To investigate expression of ryanodine receptors (RyRs) in rabbit penile corpus cavernosum smooth muscle (CCSM) cells.


METHODS
New Zealand White Rabbit CCSM cells were cultured by primary tissue culture. Using CCSM cells and fibroblast have different adherence velocity, CCSM cell can be purified. Identification of CCSM cell was by inverted microscope and immunofluorescence technique. The cDNA sequence of RyRs was found out by searching genebank. Three pair of primers were designed by Primer Premier 5.0. The RyRs subunits mRNA was detected by reverse transcription PCR in cultured CCSM cells.


RESULTS
After 7d, we found growth of cultured cells. While 15 to approximately 20 d, cells filled the bottom of culture flask. They were identified by inverted microscope and immunofluorescence technique. After purification, purity of CCSM cells was near to 100%. It suggested only RyRs1 subunit was expressive in CCSM by RT-PCR.


CONCLUSIONS
RyRsl subunit is expressed in CCSM cells. It suggests that RyRs contribute to the regulation of Ca2+ in CCSM cells.
Liu
Hong-guo
HG
Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China. Hg-liu@sohu.com
Zhu
Guang-you
GY
Zhao
Zi-qin
ZQ
Liu
Zhi-ping
ZP
D000818:Animals; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D017931:DNA Primers; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D010413:Penis / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D012333:RNA, Messenger; D019837:Ryanodine Receptor Calcium Release Channel; D002118:Calcium