Penile Nitric Oxide Synthase Research Articles

Effects of Cultured Cordycep militaris on Sexual Performance and Erectile Function in Streptozotocin-Induced Diabetic Male Rats.

Cordyceps militaris (CM), a valuable edible and medicinal fungus, has been used as traditional medicine to treat health conditions, as well as hyposexuality in Asian societies for over a century. Due to the high demand, several artificial cultivation methods have been developed for their biological activities. In this study, CM was cultured on medium that contained white rice and silkworm pupae, and the levels of cordycepin and adenosine, as well as its aphrodisiac effects in diabetes-induced erectile dysfunction (DIED), were evaluated. Diabetic rats were induced by streptozotocin (STZ) injection and administered orally with CM (0.1, 0.5, and 1.0 g/kg BW/day) for 3 weeks. Diabetic rats in negative and positive control groups received vehicle and sildenafil citrate (5 mg/kg), respectively. Results showed the changes in mating behaviour in which mount latency and intromission latency were significantly increased in diabetic rats, compared with the normal control group. Diabetic rats also showed a significant reduction in intracavernosal pressure (ICP) response to cavernous nerve stimulation, sperm count, testosterone level, penile nitric oxide synthase (NOS), and testicular superoxide dismutase (SOD) activities, when compared to the normal control group. Administration of CM (0.1, 0.5, and 1.0 g/kg BW/day) reversed the effects of diabetes on the mating behaviour, and the ICP responses to electrical stimulation. Moreover, the levels of penile NOS, testicular SOD activities, testosterone, and sperm count were significantly increased, and testicular malondialdehyde (MDA) levels were significantly decreased in these treated diabetic rats. Diabetic rats treated with sildenafil showed a significant induction in intromission frequency and NOS and SOD activities, as well as a marked increase in ICP responses. These results suggest that CCM exerts its aphrodisiac effect, possibly through activating testosterone production and suppressing oxidative stress to enhance erectile function in diabetic rats.

Effect of tadalafil on penile nitric oxide synthase and corporal smooth muscle in rats under dutasteride treatment

Aim: To investigate the effect of tadalafil in rats administered with daily dutasteride.Methods: Twenty-four Sprague–Dawley male rats were allocated to three groups as control (group C), dutasteride (group D) and dutasteride plus tadalafil (group D + T). After a month of treatment, serum samples were obtained from rats to measure dihydrotestosterone and total testosterone. Nitric oxide (NO) synthase (NOS) immunoreactivity and levels of NOS enzyme isoforms, NO and cyclic guanosine monophosphate (cGMP) were evaluated in the harvested penile tissues. Also, corporal smooth muscle and collagen were examined.Results: Staining intensities of neuronal NOS and endothelial NOS were significantly lower in group D (p < .05). They were similar between group C and group D + T. Immunoreactivity of inducible NOS was observed higher in group D than group C (p = .01) whereas group D + T had the highest iNOS (p<.001). ELISA revealed similar outcomes in terms of NOS enzyme isoform levels. The mean of smooth muscle to collagen ratio was the lowest in group D (p < .001) and it was similar among group C and group D + T (p = .072). Group D had the lowest cGMP and NO levels (p < .05) and they did not differ between group C and group D + T (p>.05). Group D and group D + T had significantly decreased dihydrotestosterone and increased testosterone, compared to group C (p < .001). They were similar between group D and group D + T.Conclusion: Daily treatment with tadalafil improves dutasteride-induced changes in rat penis.

Tadalafil Preserves Penile Nitric Oxide Synthase from Detrimental Effect of Paroxetine in Rats.

Paroxetine is a commonly prescribed SSRI that can impair erectile function in animal models via inhibition of nitric oxide synthase (NOS). Tadalafil potentiates nitric oxide (NO)-mediated responses in isolated trabecular smooth muscle and penile erection. The purpose of this study was to evaluate the impact of co-administering tadalafil with paroxetine on penile NOS levels in rats. A total of 30 male Sprague-Dawley rats were divided into 3 groups as control (Group-C), paroxetine (Group-P) and paroxetine plus tadalafil (Group-P+T). After 28 days of treatment, rats were sacrificed and their penile tissues were harvested for analysis. NOS isoform protein levels and immunoreactivity scores of NOS were assessed. Statistical significance level was set at p<0.05. Neuronal NOS (nNOS) levels were significantly decreased in group-P, compared with group-C (p<0.001). In comparison, rats in group-P+T had significantly higher nNOS levels compared to group-P (p<0.001). Endothelial NOS (eNOS) and inducible NOS (iNOS) levels were significantly higher in group-P compared with group-C (p<0.01). The levels of eNOS and iNOS in group-P+T were similar to group-C. Daily treatment with tadalafil prevented chronic paroxetine-induced changes in all three NOS isoform levels. Tadalafil treatment may therefore be a useful therapy in men with paroxetine-associated erectile dysfunction.

SaRNA Guided iNOS Up-Regulation Improves Erectile Function of Diabetic Rats

Promoter targeted saRNAs mediate sequence specific up-regulation of gene expression. We explored the therapeutic effect of RNA activation mediated iNOS gene activation on improving erectile function in a rat model of diabetes mellitus. An optimal saRNA sequence specific for iNOS promoter was cloned into an adenoviral vector, resulting in AdU6/shiNOS and AdU6/shControl. The corresponding viruses were used to transduce cultured rat cavernous smooth muscle cells. Streptozotocin induced diabetes models were established in rats and used to test the effects of intracavernous delivery of iNOS saRNA viruses on erectile function. iNOS expression in the cavernous smooth muscle cells or penile tissue of treated rats was assessed by reverse transcriptase-polymerase chain reaction and Western blot. Cyclic guanosine monophosphate was analyzed by enzyme-linked immunosorbent assay. Intracavernous pressure in response to cavernous nerve stimulation was measured using a data acquisition system on post-injection days 1, 3, 5, 7, 10 and 14. Adenovirus mediated expression of iNOS saRNA caused sustained up-regulation of iNOS in cavernous smooth muscle cells. Intracavernous injection of AdU6/shiNOS activated iNOS expression in vivo and significantly increased peak intracavernous pressure in streptozotocin induced diabetic rats via nitric oxide/intracellular cyclic guanosine monophosphate activation. Results show that saRNA mediated iNOS over expression in the penis can restore erectile function in streptozocin diabetic rats via the nitric oxide-cyclic guanosine monophosphate pathway.

Effects of chronic low- and high-dose ethanol intake on the nitrergic relaxations of corpus cavernosum and penile nitric oxide synthase in the rabbit

Epidemiological evidence showed that chronic ethanol consumption is a major risk factor in the development of impotence. The present study investigated the effects of carbachol-, electrical field stimulation (EFS)-, sodium nitroprusside (SNP)- and papaverine-induced relaxant responses in the isolated corpus cavernosum tissues from rabbits submitted to an 12-week course of chronic low (5% v/v) or high ethanol intake (30% v/v). Increased carbachol- and EFS-induced relaxant responses but not SNP and papaverine, were observed in low ethanol-fed rabbits compared with controls. However, impaired carbachol- and EFS-induced relaxant responses were observed in high ethanol-fed rabbits compared with control rabbits. There were no significant differences in SNP- and papaverine-induced relaxant responses between control and high ethanol-fed rabbits. In addition, decreased neuronal nitric oxide synthase (nNOS) and endothelial NOS (eNOS) immunoreactivity in penile tissue were found in high ethanol-fed rabbits, but increased the immunoreactivity in low ethanol-fed group, compared with control group. These results suggest that alterations in nitric oxide (NO) production within the cavernous tissue in the high ethanol-fed rabbits are, at least in part, responsible for the erectile dysfunction.

The effect of regular exercise on penile nitric oxide synthase expression in rats

Erectile dysfunction (ED) is a major public health problem that seriously affects the quality of life of patients and their partners and its prevalence increases significantly with ageing. In this study, we tested the hypothesis that age-associated decrease in penile endothelial (eNOS) and neuronal nitric oxide synthase (nNOS) activity in aged rats may be increased by regular exercise. A total of 28 young (4 m) and aged (24 m) male rats were divided into four equal groups: group 1 - young control; group 2 - young trained; group 3 - old control and group 4 - old trained group. Groups 2 and 4 rats were trained to swim for 30 min a day and 5 days a week, which lasted 8 weeks. At the end of 8 weeks, rats were sacrificed and penile tissues evaluated for eNOS and nNOS activities. eNOS and nNOS activities were evaluated by immunohistochemistry in paraffinized penile tissues and results assessed semiquantitatively. Results also were compared with healthy age-matched and adult (4 m) controls. Serum level of testosterone (T) was determined using ELISA kits (Beckman Coulter, Fullerton, CA, USA). In penile tissues of aged control rats, eNOS and nNOS staining were weakly positive; however in trained groups, eNOS and nNOS immunoreactivity were increased. In young control group, eNOS and nNOS activities were more intense than aged control. eNOS and nNOS activities were higher in adult trained group than control. Serum T concentrations were significantly higher in young and aged trained group than in control groups. We can suggest that regular exercise upregulates eNOS and nNOS expressions in the aged and young rat penis. Regular exercise may improve penile erection by increasing penile neurotransmitter in both young and aged rats.

Pubertal development of penile nitric oxide synthase (NOS)-containing nerve fibers in the rat

Objectives: To evaluate the expression of nitric oxide synthase (NOS)-containing nerve fibers in penile tissue in different age groups in the rat and to measure serum testosterone levels during this developmental process. Material and Methods: Fifteen male Sprague-Dawley (SD) rats divided into 3 equally numbered groups of different age (40 days (d), 54d and 65d) were used in this study. Penile erection was evaluated using cavernous nerve electrostimulation. Before sacrificing the rats, a penile midshaft specimen was taken for nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining and blood samples were collected to evaluate the testosterone levels. Results: Electrostimulation of the cavernous nerve revealed no significant difference in the maximal intracavernous pressure in the different age groups (44.9 ± 6.4, 54.8 ± 24.8, 45.9 ± 16.8 cm H2O, respectively; p>0.05), but the latency of the response to electrostimulation was significantly shorter in 54d and 65d than in 40d-old rats (5.4 ± 0.8 and 5.0 ± 0.7 vs. 9.0 ± 5.4 sec., respectively; p 941.5 ± 77.8 and 848.4 ± 264.3, respectively, vs. 560.0 ± 114.2 p Key words : Growth process, nitric oxide nerve fibers, penis, erection, blood testosterone, rats

Diabetes, Obesity and Erectile Dysfunction: Field Overview and Research Priorities

We provide an overview of basic, clinical and epidemiological research in the field of erectile dysfunction and important research priorities presented at the 2009 National Institute of Diabetes and Digestive and Kidney Diseases symposium on Urological Complications of Diabetes and Obesity. Experts in molecular biology, physiology, pharmacology, clinical trials, epidemiology and urological surgery highlighted current knowledge on erectile dysfunction associated with diabetes mellitus and obesity. Predictable associations between erectile dysfunction, and poor diabetic control and modifiable risk factors, including body mass index, have not yet been translated into randomized trials in the United States. The relationship between erectile dysfunction and metabolic syndrome, and surrogate markers for erectile dysfunction requires further investigation. Basic research aimed at discovering disease mechanisms and therapeutic targets has focused on autonomic neuropathy, vascular dysfunction, smooth muscle contractile function and matrix. However, significant gaps exist in regard to the integration of molecular, cellular and functional data. Animal models of type 2 diabetes and obesity associated erectile dysfunction require investigation because most basic science studies have used rodent models of type 1 diabetes. Studies are needed to synthesize a systems biology understanding of erectile function/dysfunction, and characterize and disseminate rodent models of erectile dysfunction associated with type 2 diabetes and obesity. Clinical studies are needed of promising intervention and prevention strategies. Leveraging existing and future cohort phenotypes, and biological samples is needed for risk factor analysis, biomarker discovery and genome wide association studies.

Sildenafil Inhibits Superoxide Formation and Prevents Endothelial Dysfunction in a Mouse Model of Secondhand Smoke Induced Erectile Dysfunction

We determined the effect of passive secondhand cigarette smoke on 1) erectile function in vivo, 2) molecular mechanisms involved in penile vascular function, and 3) erectile function and penile molecular signaling in the presence of phosphodiesterase type 5 inhibitor therapy. Four groups of mice were used, including group 1--controls, group 2--mice exposed to 3 weeks of secondhand smoke (5 hours per day for 5 days per week), group 3--control plus sildenafil (100 mg/kg per day) and group 4--smoke exposed plus sildenafil (100 mg/kg per day). Cavernous nerve electrical stimulation and intracavernous injection of acetylcholine were done to assess erectile function. Constitutive and inducible nitric oxide synthase activity, reactive oxygen species generation, nitrotyrosine formation and superoxide anion levels were assessed. Decreased erectile responses to cavernous nerve electrical stimulation and impaired endothelium dependent erectile responses to ACh in mice exposed to secondhand smoke were observed. Superoxide anion was increased in endothelial and corporeal smooth muscle cells of smoking mouse penises. In mice exposed to secondhand smoke constitutive nitric oxide synthase activity was decreased, and inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation increased. Sildenafil therapy restored constitutive nitric oxide synthase activity in the penis of smoking mice, decreased inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation, and improved erectile responses to cavernous nerve electrical stimulation and acetylcholine. Short-term exposure to secondhand smoke impairs erectile function through excessive penile reactive oxygen species signaling and inducible nitric oxide synthase activity. Decreased penile constitutive nitric oxide synthase activity may be attributable to the decreased endothelial nitric oxide synthase activity resulting from increased oxidative stress. Sildenafil therapy restored nitric oxide synthase activity and decreased reactive oxygen species signaling, resulting in improved erectile function.

Metformin Restores the Penile Expression of Nitric Oxide Synthase in High‐ Fat—Fed Obese Rats

Obesity is a well-known risk factor for erectile dysfunction, which is associated with reduced penile nitric oxide synthase (NOS) expression. Recently it was reported that metformin activates AMP-activated protein kinase (AMPK), which increases the expression of neuronal (n) NOS and endothelial (e) NOS. Thus, to evaluate whether metformin restores NOS expression in penile tissue, we measured penile expression of nNOS and eNOS after 4 weeks of metformin treatment (300 mg/kg/d) in 5-month-old high-fat-fed obese (HFO) rats. HFO rats have increased fat accumulation in visceral areas and marked suppression of nNOS and eNOS expression in penile tissue. However, metformin treatment decreased visceral fat deposition and restored nNOS and eNOS expression in penile tissue. The levels of AMPK and phosphorylated AMPK were also decreased in HFO rats but were subsequently elevated by metformin treatment. These results suggest that expression of NOS was suppressed by the high-fat diet but restored by metformin treatment. The effect of metformin on the expression of NOS may be associated with its activation of AMPK.

Therapeutic stimulation of penile nitric oxide synthase (NOS) and related pathways.

The advent of a novel oral treatment for erectile dysfunction appears to have reduced the urgency for seeking alternative therapeutic innovations. However, the currently available therapies can be viewed as simply palliative, requiring on-demand access and having a significant failure rate. Therefore, additional approaches for patients unresponsive to any type of medical treatment require investigation. The modulation of the synthesis of nitric oxide (NO), the main mediator of penile erection, is an attractive target for such an approach. Models for human erectile dysfunction related to aging, diabetes and endocrine disorders have been characterized in the rat. The affected animals exhibit a reduced erectile response to electrical stimulation of the cavernosal nerve and/or a loss of spontaneous erectile reflexes. In virtually all cases, the content and/or enzyme activity of penile NOS is significantly reduced, or NO synthesis may be insufficient to overcome the relatively poor compliance of the penile corpora cavernosa smooth muscle (aging). This led to the proposal that the stimulation of penile NO synthesis may be a viable therapy for erectile dysfunction. The upregulation of penile NOS activity may be achieved by: a) increase in NOS substrate concentration, b) stimulation of NOS activity through related pathways or c) blockade of endogenous NOS inhibitors. We have shown that approach a ameliorated the erectile dysfunction of aging rats by elevating intracavernosal NOS substrate through long-term oral administration of high doses of L-arginine. Evidence supporting approach b is based on our recent discovery that the main stimulating pathway for NOS, the NMDA receptor, is present in the penis. However, NMDA receptor antagonists unpredictably trigger cavernosal relaxation in vitro in a NO-independent pathway. Finally, in the case of approach c we have found that a protein inhibitor of NOS (PIN) is expressed in the corpora cavernosa. Penile NOS content may be pharmacologically increased by: d) induction of NOS expression, and e) gene therapy with NOS cDNA. Approach d applied to the penis ameliorated erectile dysfunction in aging rats, but may be clinically unfeasible because of the risk of side effects. Our laboratory cloned the inducible and neuronal NOS isoforms expressed in the penis (PiNOS and PnNOS, respectively), and showed that approach e, based on gene transfer of PiNOS into the rat corpora cavernosa improved the erectile response. Since PnNOS is a nNOS variant different from the one expressed in the CNS, gene therapy with PnNOS is promising in combination with viral vectors and tissue-specific promoters. The aim is to achieve a long-lasting increase in penile NOS that is responsive to sexual stimulation. Following validation in animal models, oral L-arginine, intracavernosal antagonists of NMDA receptor and PIN, and intracavernosal or systemic PnNOS, may eventually translate to clinical trials.

Cloning of Rat and Human Inducible Penile Nitric Oxide Synthase Application for Gene Therapy of Erectile Dysfunction

No AccessJournal of UrologyUrological Survey: Abstracts: Sexual Function and Dysfunction1 May 1998Cloning of Rat and Human Inducible Penile Nitric Oxide Synthase Application for Gene Therapy of Erectile Dysfunction H. Garban, D. Marquez, T. Magee, J. Moody, T. Rajavashisth, J.A. Rodriguez, A. Hung, D. Vernet, J. Rajfer, and N.F. Gonzalez-Cadavid H. GarbanH. Garban More articles by this author , D. MarquezD. Marquez More articles by this author , T. MageeT. Magee More articles by this author , J. MoodyJ. Moody More articles by this author , T. RajavashisthT. Rajavashisth More articles by this author , J.A. RodriguezJ.A. Rodriguez More articles by this author , A. HungA. Hung More articles by this author , D. VernetD. Vernet More articles by this author , J. RajferJ. Rajfer More articles by this author , and N.F. Gonzalez-CadavidN.F. Gonzalez-Cadavid More articles by this author View All Author Informationhttps://doi.org/10.1097/00005392-199805000-00133AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "Cloning of Rat and Human Inducible Penile Nitric Oxide Synthase Application for Gene Therapy of Erectile Dysfunction." The Journal of Urology, 159(5), p. 1778 Department of Surgery, Division of Urology, and Department of Medicine, Division of Endocrinology, UCLA School of Medicine, Harbor-UCLA Medical Center, Torrance, California.© 1998 by American Urological Association, Inc.FiguresReferencesRelatedDetails Volume 159Issue 5May 1998Page: 1778 Advertisement Copyright & Permissions© 1998 by American Urological Association, Inc.MetricsAuthor Information H. Garban More articles by this author D. Marquez More articles by this author T. Magee More articles by this author J. Moody More articles by this author T. Rajavashisth More articles by this author J.A. Rodriguez More articles by this author A. Hung More articles by this author D. Vernet More articles by this author J. Rajfer More articles by this author N.F. Gonzalez-Cadavid More articles by this author Expand All Advertisement PDF DownloadLoading ...

Cloning of rat and human inducible penile nitric oxide synthase. Application for gene therapy of erectile dysfunction.

Erectile dysfunction is mainly due to the inability of the cavernosal smooth muscle of the penis to undergo complete relaxation. In the aging rat model, erectile dysfunction is accompanied by a reduction of penile smooth muscle compliance and, in very old animals, by a decrease in penile nitric oxide synthase (NOS), which is responsible for the synthesis of the mediator of penile erection, nitric oxide (NO). We have investigated whether the stimulation of penile NOS expression by local induction or gene therapy can mitigate erectile dysfunction in the aged rat. A mix of iNOS (inducible NOS) inducers was continuously delivered to the penises of 5- ("adult"), 20- ("old"), and 30- ("very old") mo-old rats for 3-6 days, and the erectile response to electrical field stimulation of the cavernosal nerve was measured. The erectile dysfunction observed in old and very old rats as compared to adult animals was ameliorated by treatment with iNOS inducers. Penile iNOS was detectable in the penis of these rats by Western blot, NADPH diaphorase, and NOS activity assays. Inducible NOS was inducible in vitro in both rat and human corpora cavernosal tissue and in rat penile smooth muscle cells (RPSMC), as shown by Western blots. However, NO synthesis in cavernosal tissue upon iNOS protein induction remained low, indicating that the increased NOS levels were under physiological control. The iNOS cDNA was cloned from induced RPSMC mRNA and generated by reverse transcriptase polymerase chain reaction (RT-PCR) from induced human penile smooth muscle cells and corporal tissue. The coding regions from both the rat (RPiNOS) and human (HPiNOS) penile iNOS showed several amino acid differences from their analogous isoform in nonpenile tissues. RPiNOS cDNA injected into the penis mitigated the aging-associated erectile dysfunction. The iNOS construct was detected in cavernosal tissue by PCR, and its expression by RT-PCR and Western blots. These results open the way for the possible use of NOS isoforms in the management of erectile dysfunction.

Effect of Long-term Passive Smoking on Erectile Function and Penile Nitric Oxide Synthase in the Rat

Given that smoking is a risk factor for erectile dysfunction, this study aimed to determine, in a rat model, whether long-term exposure to cigarette smoke impairs nitric oxide (NO)-dependent erectile function and reduces penile nitric oxide synthase (NOS), and if these changes are accompanied with effects on the systemic blood pressure. Adult (5 month) and old (20 month) rats were exposed to daily passive smoking for 8 wks. Three days after the conclusion of exposure, half of the animals were submitted to electrical field stimulation (EFS) of the cavernosal nerve and the maximum intracavernosal pressure (MIP) and mean arterial pressure (MAP) were determined and expressed as mm. Hg. On the other half of the animals, NOS activity in the penile cytosol was measured by the arginine/citrulline assay, and neuronal NOS (nNOS) and endothelial NOS (eNOS) contents were estimated by western blot and densitometry. When compared to controls, the smoking rats had a higher MAP in both the adult (115 vs 162) and old (113 vs 140) rats, but surprisingly the MIP also increased, from 78 to 111 (adult rats) and from 59 to 83 (old rats). Smoking reduced penile NOS activity by 73% (adult rats), and 62% (old rats), and nNOS content by 43% and 50%, respectively. In contrast, eNOS was not affected. Nitrite release, in vitro, by cavernosa slices or in rat penile smooth muscle cells (RPSMC) was not inhibited by nicotine or cotinine. These results indicate that chronic smoking in the rat leads to age-independent moderate hypertension and considerable decreases in penile NOS activity and nNOS content, that are not reflected in a reduction of the erectile response to EFS or accompanied by a decrease in penile eNOS.

Effect of Long-term Passive Smoking on Erectile Function and Penile Nitric Oxide Synthase in the Rat

Effect of Long-term Passive Smoking on Erectile Function and Penile Nitric Oxide Synthase in the Rat

Androgen and pituitary control of penile nitric oxide synthase and erectile function in the rat.

Castration of adult male rats reduces by half the penile erectile response to electrical field stimulation (EFS) of the cavernosal nerve, and the activity of penile nitric oxide synthase (NOS). Both changes are prevented by androgen administration. We have now investigated whether other strategies of androgen ablation or competition may act as stronger inhibitors, and, if so, whether the stronger inhibition is due to the depletion of penile NOS content. Rats were castrated or left intact and were treated daily as follows: 1) intact, with the antiandrogen flutamide (25 mg/kg/day, i.p.); 2) castrated, with similar treatment; 3) castrated, with 17 beta-estradiol 3-benzoate (estradiol; via silastic tubing, s.c.). Additional groups of intact rats received injections of a GnRH antagonist (GnRHA, 1.25 mg/kg, s.c.), or were hypophysectomized and left untreated. Controls were untreated intact and castrated animals. After 7 days, rats were subjected to EFS, and the ratios between maximal intracavernosal pressure (MIP) and mean arterial pressure (MAP) were measured. Penile NOS activity and the contents of neuronal NOS (nNOS) and endothelial NOS (eNOS) were determined. Castration reduced the MIP:MAP ratio and penile NOS activity. Androgen receptor blockade with flutamide induced a similar response in intact rats. When flutamide treatment was combined with castration, the erectile response was nearly abolished, but NOS activity was not decreased below the values in castrated rats. Estradiol given to castrated rats and hypophysectomy or GnRHA treatment in intact rats diminished the erectile response below the level in castrated animals. In hypophysectomized rats, penile NOS activity fell below levels in castrated animals. contents of nNOS and eNOS were not significantly reduced by any treatment. These data suggest that penile erection in the rat is completely dependent on androgens, presumably because of their role in the maintenance of penile NOS activity and of other ancillary factors. However, only the complete blockade of residual androgen effects at the tissue level or a total androgen depletion can abolish the erectile response.

Cavernosal nerve stimulation in the rat reverses castration-induced decrease in penile NOS activity.

Castration in the rat reduces both the erectile response and penile nitric oxide synthase (NOS) activity, and these effects are prevented by androgen administration. In this study we determined that the decrease of penile NOS in the castrated rat is due to NOS enzyme inhibition rather than to a reduction of its content and that this inhibition may be reversed. Adult rats were either castrated or left intact, and, after 1 wk, electrical field stimulation (EFS) was applied to the cavernosal nerve and the penises were excised either at the peak of erection or after detumescence. Penile NOS activity in the non-EFS castrates decreased by 70% as compared with the intact non-EFS-treated controls, but neuronal NOS content remained unaffected despite changes in regional distribution. NOS activity in the castrated penis was restored to normal values by EFS at the peak of stimulation, and then decreased on detumescence. This was in contrast to the intact controls in which EFS did not stimulate penile NOS activity. These data indicate that androgen depletion in the rat reduces penile NOS activity rather than NOS content and that this enzyme inhibition is reversed by cavernosal nerve stimulation.

Reduction of penile nitric oxide synthase in diabetic BB/WORdp (type I) and BBZ/WORdp (type II) rats with erectile dysfunction.

Erectile dysfunction occurs frequently in human diabetes, and it is sometimes associated with hypogonadism. These conditions also develop in a model of insulin-dependent (type I) diabetes, the BB/WORdp (diabetic prone) rat but have not yet been investigated in the model of insulin-resistant (type II) diabetes, the BBZ/WOR rat. It is also unknown whether diabetes-related impotence is due to reduced levels of the mediator of penile erection, nitric oxide, caused by a decrease of nitric oxide synthase (NOS) in the penis. To clarify these questions, groups (n = 5-6) of diabetic BB/WORdp (insulin-maintained) and BBZ/WOR rats were age-matched with diabetic-resistant BB/WORdr and non-diabetic BB/WORdp rats and submitted to determinations of serum glucose, testosterone, and penile reflexes (cups and flips). Erectile dysfunction was found in all of type I and in most of type II diabetic animals (glycemias of 25.0 and 31.1 mM), at the selected mean ages of 310 and 180 days old, respectively. This was evidenced by over 95% decreases of erectile reflexes in both types of diabetes and was accompanied by 75% reduction of serum testosterone. Soluble NOS activity was measured in penile tissue from the diabetic rats with impaired erectile reflexes and in the corresponding controls, by the (3H)-L-arginine/citrulline conversion assay. The neuronal NOS isoform (nNOS) content was determined by a semiquantitative western blot assay. Both types of diabetes showed a marked decrease of penile NOS activity (74 and 55%, respectively), and a lower reduction of penile nNOS content (47 and 33%, respectively). No endogenous NOS inhibitor was detected in the diabetic type I penile cytosol by cross-mixing NOS activity assays. Our data support a common etiology for erectile dysfunction present in rats with types I and II diabetes mellitus and suggest that the etiology is related to a decrease of penile NOS derived in part from serum androgen deficiency.

Reduction of penile nitric oxide synthase in diabetic BB/WORdp (type I) and BBZ/WORdp (type II) rats with erectile dysfunction

Erectile dysfunction occurs frequently in human diabetes, and it is sometimes associated with hypogonadism. These conditions also develop in a model of insulin-dependent (type I) diabetes, the BB/WORdp (diabetic prone) rat but have not yet been investigated in the model of insulin-resistant (type II) diabetes, the BBZ/WOR rat. It is also unknown whether diabetes-related impotence is due to reduced levels of the mediator of penile erection, nitric oxide, caused by a decrease of nitric oxide synthase (NOS) in the penis. To clarify these questions, groups (n = 5-6) of diabetic BB/WORdp (insulin-maintained) and BBZ/WOR rats were age-matched with diabetic-resistant BB/WORdr and non-diabetic BB/WORdp rats and submitted to determinations of serum glucose, testosterone, and penile reflexes (cups and flips). Erectile dysfunction was found in all of type I and in most of type II diabetic animals (glycemias of 25.0 and 31.1 mM), at the selected mean ages of 310 and 180 days old, respectively. This was evidenced by over 95% decreases of erectile reflexes in both types of diabetes and was accompanied by 75% reduction of serum testosterone. Soluble NOS activity was measured in penile tissue from the diabetic rats with impaired erectile reflexes and in the corresponding controls, by the (3H)-L-arginine/citrulline conversion assay. The neuronal NOS isoform (nNOS) content was determined by a semiquantitative western blot assay. Both types of diabetes showed a marked decrease of penile NOS activity (74 and 55%, respectively), and a lower reduction of penile nNOS content (47 and 33%, respectively). No endogenous NOS inhibitor was detected in the diabetic type I penile cytosol by cross-mixing NOS activity assays. Our data support a common etiology for erectile dysfunction present in rats with types I and II diabetes mellitus and suggest that the etiology is related to a decrease of penile NOS derived in part from serum androgen deficiency.