title: Smooth muscle myosin expression, isoform composition, and functional activities in rat corpus cavernosum altered by the streptozotocin-induced type 1 diabetes.
abstract: Diabetes mellitus (DM) is a quite common chronic disease, and the prevalence of erectile dysfunction (ED) is three times higher in this large population. Although diabetes-related ED has been studied extensively, the actin-myosin contractile apparatus was not examined. The mRNAs encoding smooth muscle myosin (SMM) heavy chains (MHC) and essential light chains (LC(17)) exist as several different alternatively spliced isoforms with distinct contractile properties. Recently, we provided novel data that blebbistatin (BLEB), a specific myosin II inhibitor, potently relaxed corpus cavernosum smooth muscle (CCSM). In this study, we examine whether diabetes alters SMM expression, alternative splicing, and/or functional activities, including sensitivity to BLEB. By using streptozotocin (STZ)-induced 2-mo diabetic rats, functional activities were tested in vivo by intracavernous pressure (ICP) recording during cavernous nerve stimulation and in vitro via organ bath contractility studies. SMM isoform composition was analyzed by competitive RT-PCR and total SMM, myocardin, and embryonic SMM (SMemb) expression by real-time RT-PCR. Results revealed that the blood glucose level of STZ rats was 407.0 vs. 129.5 mg/dl (control). STZ rats exhibited ED confirmed by significantly increased CCSM contractile response to phenylephrine and decreased ICP response. For STZ rats, SM-B, LC(17a) and SM2 isoforms, total SMM, and myocardin expression increased, whereas SM-A, LC(17b), and SM1 isoforms were decreased, with SMemb unchanged. BLEB was significantly more effective in relaxing STZ CCSM both in vitro and in vivo. Thus we demonstrated a novel diabetes-specific effect on alternative splicing of the SMM heavy chain and essential light chain genes to a SMM isoform composition favoring a heightened contractility and ED. A switch to a more contractile phenotype was supported further by total SMM expression increase. Moreover, the change in CCSM phenotype was associated with an increased sensitivity to BLEB, which may serve as a novel pharmacotherapy for ED.
authors.affiliation: Department of Urology, Albert Einstein College of Medicine, Bronx, NY, USA.
authors.forename: Xinhua
authors.initials: X
authors.lastname: Zhang
authors.forename: Nirmala D
authors.initials: ND
authors.lastname: Kanika
authors.forename: Arnold
authors.initials: A
authors.lastname: Melman
authors.forename: Michael E
authors.initials: ME
authors.lastname: DiSanto
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D003922:Diabetes Mellitus, Type 1 / Q000150:complications / Q000378:metabolism* / Q000503:physiopathology*; D004351:Drug Resistance; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007172:Erectile Dysfunction / Q000150:complications / Q000188:drug therapy / Q000378:metabolism* / Q000503:physiopathology*; D005786:Gene Expression Regulation; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism; D024462:Myosin Type II / Q000037:antagonists & inhibitors; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010413:Penis / Q000187:drug effects / Q000294:innervation / Q000378:metabolism* / Q000503:physiopathology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D024745:Smooth Muscle Myosins / Q000235:genetics / Q000378:metabolism*; D013311:Streptozocin / Q000633:toxicity; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D000099251:Myocardin
chemical_list: D004791:Enzyme Inhibitors; D006576:Heterocyclic Compounds, 4 or More Rings; D018995:Myosin Heavy Chains; D018994:Myosin Light Chains; D024462:Myosin Type II; D009687:Nuclear Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D024745:Smooth Muscle Myosins; D013311:Streptozocin; D015534:Trans-Activators; D000099251:Myocardin; C472645:blebbistatin