Expression of nitric oxide-sensitive guanylyl cyclase subunits in human corpus cavernosum.
The muscles of the corpus cavernosum of the penis relax in response to stimulation of non-adrenergic, non-cholinergic nerves or nitric oxide (NO)-donating drugs to elicit erection. It is generally assumed that NO mediates this effect via activation of soluble guanylyl cyclase and a subsequent increase in cyclic guanosine 3', 5'-monophosphate concentration. However, there are no data on the expression of this enzyme in human corpus cavernosum. The purpose of the present study was the molecular characterization of NO-sensitive guanylyl cyclase in human corpus cavernosum. RNA was extracted from tissue samples obtained from seven patients undergoing penile prosthetic surgery or correction of penile deviation. Reverse transcriptase-polymerase chain reaction (RT-PCR) with specific primers for the subunits of NO-sensitive guanylyl cyclase was performed, and PCR products were subcloned and sequenced. Specific amplification products encoding the alpha(1), beta(1), alpha(2), and beta(2) subunits were detected. In addition, we isolated a transcript encoding a novel variant beta(2) subunit. To test whether this novel transcript arises by alternative splicing or whether it is encoded by a separate gene, a 4000-bp clone of the corresponding genomic DNA sequence was isolated. Sequence analysis suggests that the novel beta(2) variant arises by alternative splicing from the same gene as the beta(2) subunit on chromosome 13. In conclusion, our findings suggest the presence of different subunit mRNAs of NO-sensitive guanylyl cyclase in human corpus cavernosum.
Behrends
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S
Institut für Experimentelle und Klinische Pharmakologie, Universität Hamburg, D-20251, Hamburg, Germany. behrends@plexus.uke.uni-hamburg.de
Steenpass
A
A
Porst
H
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Scholz
H
H
D000595:Amino Acid Sequence; D006162:Guanylate Cyclase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000378:metabolism*; D010413:Penis / Q000201:enzymology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D009569:Nitric Oxide; D006162:Guanylate Cyclase