Arginine Metabolism: Enzymology, Nutrition, and Clinical Signicance

Role of Arginase in the Male and Female Sexual Arousal Response1,2

Noel N. Kim,* David W. Christianson,** and Abdulmaged M. Traish*3
*Department of Urology and Institute for Sexual Medicine, Boston University School of Medicine, Boston, MA 02118; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118; and **Roy and Diana Vagelos Laboratories, Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104
ABSTRACT The NO-cGMP pathway plays a key role in the male and female genital sexual arousal response. Nitric oxide synthase (NOS) utilizes L-arginine and oxygen as substrates to produce nitric oxide (NO) and citrulline. Arginase is a metalloenzyme that catalyzes the hydrolysis of L-arginine to produce L-ornithine and urea. It is proposed that arginase competes for L-arginine and reduces NOS activity in genital tissues, thus modulating sexual function. Using 2 transition state analogue inhibitors of arginase, 2(S)-Amino-6-boronohexanoic acid (ABH) and S-(2-boronoethyl)-L-cysteine (BEC), we have characterized arginase activity in penile and vaginal tissue. Neither of these inhibitors has activity against NOS. Thus, ABH and BEC are useful compounds for examining the role of arginase in genital tissue physiology, without directly inuencing NOS activity. We present data to suggest that arginase may regulate NO production by competing for endogenous pools of L-arginine. In this fashion, arginase is an indirect regulator of penile and vaginal blood ow and specic arginase inhibitors may improve genital blood ow during sexual arousal. As evidenced by the upregulation of arginase in specic disease states, its distribution in the vagina, and its modulation by sex steroid hormones, this enzyme may also participate in numerous other physiological and pathophysiological processes, such as tissue growth, brosis, and immune function. J. Nutr. 134: 2873S2879S, 2004. KEY WORDS:

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arginase

arginine

nitric oxide synthase

erectile function

genital hemodynamics

Sexual dysfunction is a common and signicant medical problem that adversely affects health, well-being, quality of life, and interpersonal relationships (13). Although multiple categories of sexual dysfunction have been dened, this article will limit its discussion to genital arousal disorders as it pertains to penile erectile dysfunction in men and vaginal and clitoral engorgement insufciency in women. Male erectile dysfunction is dened as the persistent inability to achieve or maintain an erection for satisfactory sexual performance. The prevalence of erectile dysfunction increases with age, doubling between the ages of 40 and 70 y for moderate erectile dysfunction and tripling over the same 3-decade time period for complete dysfunction (4). Although age is a signicant correlate, other factors such as treated diabetes mellitus, heart disease and hypertension, medications for diabetes, cardiovascular disease, and diminished values of highdensity lipoproteins also predict erectile dysfunction (4). Research on the biochemical and physiological mechanisms
1 Prepared for the conference Symposium on Arginine held April 5 6, 2004 in Bermuda. The conference was sponsored in part by an educational grant from Ajinomoto USA, Inc. Conference proceedings are published as a supplement to The Journal of Nutrition. Guest Editors for the supplement were Sidney M. Morris, Jr., Joseph Loscalzo, Dennis Bier, and Wiley W. Souba. 2 This work was supported by NIH grants DK56846 (AMT), DK02696 (NNK) and GM49758 (DWC) from the National Institute of Diabetes and Digestive and Kidney Diseases and the National Institute of General Medical Sciences. 3 To whom correspondence should be addressed. E-mail: atraish@bu.edu.

regulating erectile tissue trabecular smooth muscle contractility has led to signicant advances in the pharmacological management of erectile dysfunction (5 8). The female genital arousal response is manifested by attaining and maintaining sufcient sexual excitement leading to genital engorgement, swelling, and lubrication. Genital vasocongestion and vaginal lubrication responses result from increased blood ow to the genital tissues and production of lubricating uid transudate from the vaginal epithelium. In contrast to the physiology of penile erection, there is limited understanding of local regulatory mechanisms modulating clitoral and vaginal smooth muscle tone and how these mechanisms are altered by changes in the hormonal milieu and disease states. Women with sexual arousal disorder have sexual complaints of diminished vaginal lubrication, increased time for arousal, diminished vaginal and clitoral sensation, and difculty with orgasm. Based on the National Health and Social Life Survey, one-third of women report lack of sexual interest, one-fourth report orgasmic problems, one-fth report lubrication difculties, and the same percentage nd sex not pleasurable (2,3). The physiology of vaginal hemodynamics and lubrication responses is highly dependent on tissue structural and functional integrity and involves complex neurovascular processes modulated by various local neurotransmitters (9,10), vasoactive agents, sex steroid hormones, and growth factors. Studies in both male and female genital tissues indicate

0022-3166/04 $8.00 2004 American Society for Nutritional Sciences.

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that the nitric oxide (NO)4/guanosine-3,5-cyclic monophosphate (cGMP) pathway is an important regulator of blood ow and engorgement during sexual arousal. L-arginine serves as a critical substrate for the generation of NO by nitric oxide synthase (NOS). Arginine is also utilized by arginase and converted to ornithine and urea. The key role of arginine as a substrate for both nitric oxide synthase and arginase serves as a potential point of regulation for the NO/cGMP pathway. This review summarizes the available data on arginase in male and female genital tissues and its regulatory role in the sexual arousal response. Role of nitric oxide and arginase in male sexual arousal function Regulation of penile tumescence by nitric oxide. During erection, the penis acts as a capacitor, accumulating blood under pressure (5 8). Dilation of the resistance arterial bed of the penis provides ow and pressure to the erectile bodies (corpora cavernosa) and relaxation of the trabecular smooth muscle allows expansion of the lacunar spaces and trapping of blood by stretching and compression of the draining venules. When the trabecular smooth muscle of the corpora cavernosa is fully relaxed, the intracavernosal pressure is dependent on the cavernosal arterial pressure (57). The state of relaxation or contraction of the arteriolar and trabecular smooth muscle determines penile erection or accidity. It is generally accepted that under normal physiological conditions, sexual stimulation causes the synthesis and release of nitric oxide from penile nonadrenergic, noncholinergic (NANC) nerves and the endothelium lining the lacunar spaces (1117). The 3 main types of NOS (neural, endothelial, and inducible) have been identied in penile corpus cavernosum. Each type utilizes L-arginine as a cosubstrate with molecular oxygen to generate NO and citrulline. Activation of neurogenic and endothelial NO synthases results in production of NO that diffuses into smooth muscle cells of the resistance arteries and erectile tissue and binds to the heme component of soluble guanylyl cyclase, stimulating the synthesis of cGMP (Fig. 1). Binding of cGMP to cGMP-dependent protein kinases (PKG) or cGMP-dependent ion channels results in reduction of intracellular calcium, via calcium sequestration and extrusion, and activation of myosin light chain phosphatases, which inhibits smooth muscle contraction and enhances penile erection. It should be noted that NO may also mediate smooth muscle relaxation independent of cGMP. In aortic smooth muscle cells NO has been shown to directly activate Ca2-dependent K channels (18). In human corpus cavernosum smooth muscle cells, NO was shown to directly activate sodium-potassium ATPase to cause hyperpolarization (19). An important factor underlying the pathology of erectile dysfunction may be attenuation in NO/cGMP signaling (17,20,21). NO production in penile tissue is reduced in aging (2123) and diabetes (24 28), conditions having a high degree of association with erectile dysfunction. Several reports suggested that decreased NO production due to endothelial dysfunction or nerve injury may represent a central mechanism of erectile dysfunction. Reduced production of NO by

FIGURE 1 Key reactions in the NO/cGMP pathway: Reactions 1 and 2 are expected to enhance the relaxation of smooth muscle since the end product is cGMP. Reactions 3, 4, and 5 are expected to diminish the relaxation response since they impede NO synthesis, scavenge NO or hydrolyze cGMP, respectively.

4 Abbreviations used: ABH, 2(S)-amino-6-boronohexanoic acid; BEC, S-(2boronoethyl)-L-cysteine; cGMP, guanosine-3,5-cyclic monophosphate; DFMO, diuoromethylornithine; DHEA, dehydroepiandrosterone; DHT, dihydrotestosterone; mRNA, messenger RNA; NANC, nonadrenergic, noncholinergic; NO, nitric oxide; NOHA, N-hydroxy-L-arginine; NOS, nitric oxide synthase; PKG, cGMPdependent protein kinase; TGF-beta, transforming growth factor beta.

the endothelium may be the result of decreased NOS protein expression and activity, increased NO scavenging, increased endogenous inhibitors of NOS, or decreased levels of substrate (L-arginine and oxygen) and cofactors. Effects of exogenous L-arginine on erectile function. Several studies in human and animal subjects have investigated the effects of exogenous L-arginine on restoring NOS activity and improving erectile function (29 34). In a controlled crossover study, oral L-arginine at a dose of 500 mg, given 3 times per day, did not improve erectile function when compared to placebo (32). In contrast, Chen et al. (31) reported that oral administration of L-arginine in high doses (5 g/d) caused signicant subjective improvement in sexual function in men with organic erectile dysfunction only if they had decreased NOx excretion or production prior to arginine supplementation. Interestingly, objective measures such as penile hemodynamics were not affected by oral L-arginine. In a double blind, placebo controlled, 3-way crossover, randomized clinical trial, Lebret et al. (30) compared the efcacy and safety of 6 g of L-arginine glutamate and 6 mg of yohimbine hydrochloride on treatment of erectile dysfunction. The results of these studies showed that on demand (12 h before intended sexual intercourse) oral administration of L-arginine glutamate and yohimbine combination is effective in improving erectile function in patients with mild to moderate erectile dysfunction. However, the data from these studies remain equivocal in determining whether dietary L-arginine improves erectile function in men with erectile dysfunction. In animal studies, Moody et al. (33) demonstrated that long-term oral administration of supraphysiological doses of L-arginine improves erectile function in the aging rat. Yildirim et al. (34) showed that administration of oral L-arginine to diabetic rabbits increases endothelium-dependent relaxation of rabbit corpus cavernosum but had no effect on neurogenic relaxation in diabetic animals. Angulo et al. (35) investigated the effects of phentolamine and L-arginine on neurogenic relaxation of healthy rabbit corpus cavernosum in organ bath studies. The authors concluded that a synergistic interaction between alpha adrenergic blockade and the potentiation of the NO/cGMP pathway increases neurogenic relaxation of corpus cavernosum in vitro. L-arginine alone was without effect in potentiating nerve-mediated relaxation. In a subsequent study, Angulo et al. (36) examined the effects of L-arginine and NG-hydroxy-L-arginine on corpus cavernosum relaxation in young and old animals. Although addition of L-arginine was unable to relax phenylephrine-contracted cavernosal tissue strips, addition of NG-hydroxy-L-arginine relaxed the tissue strips effectively and elevated tissue levels of cGMP. The authors concluded that addition of NG-hydroxy-L-arginine improved NOS activity while administration of exogenous Larginine did not. Since hydroxyarginine is a substrate for NOS and an inhibitor of arginase, it remains unclear whether the

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effects observed are solely due to NOS activation or a combination of NOS activation and arginase inhibition. In nongenital vascular tissues, L-arginine restores endothelial function in hypercholesterolemic animals by enhancing NO production and by protecting NO from early breakdown by superoxide. Further, dietary L-arginine reduces the progression of atherosclerosis and vascular oxidative stress and preserves endothelial function in cholesterol-fed rabbits (37 44). These data suggest that hypercholesterolemia-induced endothelial dysfunction can be reversed by L-arginine administration. Similarly, L-arginine enhances acetylcholine-induced relaxation in aortic ring preparations from diabetic rats (45). L-arginine availability for NOS. Bo ger et al. (39) have suggested that the concentration of L-arginine circulating in the blood approximates 100 mol/L. Other studies have indicated that the intracellular concentrations of L-arginine in endothelial cells range from 100 to 800 mol/L (46 49). The Michaelis-Menten constant (Km) for L-arginine binding to NOS is in the range of 210 mol/L (50 52). Based on this information, one would expect that various NOS isoforms are saturated with L-arginine at the physiological concentrations and that exogenous L-arginine administration or arginase inhibition should have little or no effect on NOS activity. However, a number of studies using in vivo and in vitro experiments indicated that NOS activity was increased by exogenous L-arginine, in spite of the high intracellular concentrations (42,44,5357). On the contrary, studies by Muggi & Harrison (58) and Angulo et al. (36) have indicated that exogenous L-arginine had no effect on acetylcholine or EFSmediated responses. The potential reasons for discrepancies between various clinical and laboratory studies on the effects of exogenous L-arginine and alternative mechanisms have been elegantly discussed by Loscalzo (59). Alternative mechanisms including compartmentalization of L-arginine into various intracellular pools have also been discussed (60). This hypothesis is under investigation by a number of laboratories and available data suggest that competition for intracellular L-arginine by arginase and NOS may be more important in regulating NOS activity than the overall intracellular levels of L-arginine (56,57,61 64). In genital tissues, we propose that reduced L-arginine availability to NO synthase and decreased production of NO would lead to attenuated vascular and trabecular smooth muscle relaxation and erectile dysfunction (Fig. 1). Arginase activity in penile corpus cavernosum. Our studies indicate that messenger RNA (mRNA) for both isoforms of arginase I and II can be detected by RT-PCR in tissue samples of human penile corpus cavernosum (65). We have also demonstrated that arginase enzyme activity is present in tissue extracts of rabbit and human corpus cavernosum, as assessed by conversion of [14C]L-arginine to urea and ornithine (65,66). Limited evidence exists to suggest that alterations in arginase levels may contribute to various pathological states. Bivalacqua et al. (26,27) have investigated changes in arginase mRNA and protein and enzyme activity in diabetic and normal human cavernosal tissues. Corpus cavernosum from diabetic patients with erectile dysfunction had higher levels of arginase II mRNA, protein, and enzymatic activity than nondiabetics. In contrast, mRNA and protein levels for arginase I isoform were not signicantly different in diabetic and nondiabetic cavernosal tissue. Thus, increased expression of arginase II in diabetic cavernosal tissue may contribute to erectile dysfunction associated with diabetes. In addition to perturbations in smooth muscle tone, arginase may directly or indirectly regulate trophic processes within penile tissue. For example, injection of TGF-beta in the rat penis induces localized

brosis and signicantly increases expression of arginase II protein (26,27). The authors postulated that this elevation in arginase II may attenuate the synthesis of NO by eNOS and result in eNOS downregulation in this disease condition. Inhibition of arginase activity modulates smooth muscle contractility and erectile function. The use of specic arginase inhibitors has been particularly helpful in elucidating the regulatory role of arginase. Two such inhibitors are (S)-2amino-6-boronohexanoic acid (ABH) and S-(2-borono-ethyl)-L-cysteine (BEC), which have no inhibitory activity against NOS (65,66). Both ABH and BEC contain trigonal planar boronic acid side chains that are isosteric with the trigonal planar guanidinium group of L-arginine. Since the boronic acid moiety is highly electrophilic, it undergoes nucleophilic attack by a hydroxide ion that bridges the 2 manganese ions in the arginase active site. This reaction results in the formation of a tetrahedral boronate anion that mimics the tetrahedral intermediate and its anking transition states in the arginase mechanism. Because ABH and BEC are designed as substrate analogues, they react in the arginase active site to bind as transition state analogues, and this is the origin of the high afnity of these inhibitors. ABH is the most potent arginase inhibitor known to date, with Ki 0.1 mol/L against rat arginase I (67 69) and Ki 8.5 nmol/L against human arginase II (70). The rst use of a tight-binding boronic acid-based arginase inhibitor to modulate an NO-dependent process was in the study of the internal anal sphincter muscle of the adult opossum (69). Arginase attenuated internal anal sphincter muscle relaxation triggered by NANC-nerve stimulation, and this effect was reversed by the arginase inhibitor ABH. Moreover, the use of ABH alone augmented NANC nerve- mediated relaxation of the internal anal sphincter muscle, and this augmentation was 250 times more potent than that achieved with N-hydroxy-L-arginine (NOHA). These results demonstrated the role of arginase and the potential utility of an arginase inhibitor in regulating smooth muscle tone. Comparable results were obtained in studies of smooth muscle relaxation in penile corpus cavernosum. In isolated strips of penile erectile tissue, neurogenic relaxation was enhanced by addition of 0.1, 0.5, or 1 mmol/L of either arginase inhibitor (65,66). The enhancement in smooth muscle relaxation was more pronounced at the lower frequencies. The data from these studies suggest that the neurogenic relaxation in penile erectile tissue, which is known to be mediated by NO, was potentiated by inhibition of arginase. Endothelium-dependent relaxation has also been shown to be regulated by arginase. Utilizing the arginase inhibitor, Nhydroxy-nor-L-arginine (nor-NOHA), Masuda et al. (71) have shown that addition of 0.1 mmol/L of nor-NOHA facilitated endothelium-dependent relaxation and this was reversed by addition of the NOS inhibitor L-NAME. In vivo studies with arginase inhibitors have proven to be consistent with the in vitro studies, conrming the physiological role of arginase in regulating NO synthesis. Intracavernosal administration of ABH in the anesthetized rabbit facilitated penile erection elicited by pelvic nerve stimulation (Fig. 2) (72). These observations suggest that inhibition of arginase facilitated erections by increasing the compartmentalized pool of arginine as a substrate for neural and endothelial NOS. The benecial effects of specic arginase inhibitors may become even more noticeable when they are used in pathological conditions where arginase levels may be signicantly increased. For instance, the reduced NOS activity observed in penile erectile tissue from diabetic men was normalized when treated with the arginase inhibitor ABH (26,27). Although

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FIGURE 2 Effects of ABH administration on penile erection: Changes in penile intracavernosal pressure were recorded in response to pelvic nerve stimulation in anesthetized male New Zealand White rabbits. The pelvic nerve was electrically stimulated 10 min after intracavernosal injection of vehicle (control; 40% propylene glycol) or 150 g of ABH. Representative pressure recordings are shown in the top panels. Amplitude, duration, and area-under-the-curve (AUC) were determined for each response and the means SEM are shown for the relevant parameters in the 2 lower graphs (n 4; *P 0.05 compared to control). Reprinted with permission from Cama et al. (72), Copyright 2003, American Chemical Society.

studies on arginase in disease models of genital organs are lacking, data from nongenital vascular tissues are accumulating in the literature. Osanai et al. (73) and Shukla et al. (74) have suggested that age modies the amino acid pool size and L-arginine availability. Studies in young and adult rats (56) showed that inhibition of arginase activity with BEC, Nhydroxy-nor-L-arginine (nor-NOHA) or diuoromethylornithine (DFMO) produced vasodilation in aortic rings. In aortic tissue from old rats, arginase activity and expression was increased while BEC and DFMO restored NOS activity and cGMP levels when compared to aortic tissue from young rats. In porcine coronary microvessels, Zhang et al. (75) have shown that the endothelium expresses arginase and inhibition of arginase activity with DFMO stimulates NO production and enhances vasodilation. Role of NO and arginase in female sexual arousal function A developing hypothesis is that following sexual stimulation, impaired genital arousal, as manifested by inadequate clitoral and/or vaginal engorgement, is linked in part to diminished arterial inow. In the female animal model, atherosclerosis of the ilio-hypogastric-pudendal arterial bed resulted

in diminished vaginal and clitoral arterial blood inow as well as reduced vaginal and clitoral tissue pressure responses following pelvic nerve stimulation compared to healthy control animals (76). Further, the NO/cGMP pathway is an important modulator of vaginal smooth muscle contractility (77,78) and blood ow in the animal model (79,80). Expression of arginase activity in the female genital tissues. We have reported that NOS and arginase are differentially distributed among the anatomic regions of the rabbit vagina (81). Total activity of nitric oxide synthases (nNOS & eNOS) was higher in the proximal than distal vagina. However, arginase activity was greater in the distal than proximal vagina (81). The physiological signicance for this unique regional anatomical distribution of these enzymes in this animal model is unknown at present. The distribution of NOS and arginase in the human vagina remains poorly characterized. We have also reported that ovariectomy enhanced total NOS activity in the proximal vagina with concomitant decrease in arginase activity in the proximal and distal vagina, suggesting differential regulation of these enzymes by sex steroid hormones in the various anatomical regions of the rabbit vagina (81). Interestingly, vaginal and clitoral arginase activity was reduced by ovariectomy and upregulated by estrogen replacement in the distal vagina, suggesting that arginase is under regulation by estrogen hormones (81,82). Treatment of ovariectomized animals with DHEA, 5-androstenediol, or 5DHT did not signicantly change arginase activity in the vagina relative to ovariectomized animals treated with vehicle. Treatment of ovariectomized animals with estradiol in combination with testosterone or progesterone increased arginase activity in the clitoris and distal vagina. These observations suggest that, in vaginal and clitoral tissues, arginase activity is modulated by estrogens in a tissue- and region-specic manner. The physiological signicance of arginase regulation by sex steroid hormones in the vagina and clitoris may have metabolic and functional roles; however, the role of arginase has yet to be investigated in vaginal and clitoral tissues. We suggest that arginase plays a critical role in vaginal epithelium growth in response to steroid hormones, via synthesis of polyamines via a pathway similar to that reported in endothelial cells (83,84). It is possible that arginase plays a regulatory role in endothelial proliferation in the vascular bed of the clitoris. However, this hypothesis has not yet been tested in vaginal epithelium or clitoral tissues in vitro or in vivo. Inhibition of arginase activity enhances female genital hemodynamics. Previous studies in the female rabbit have suggested that the NO-cGMP pathway is a modulator of vaginal vasocongestion and may play a role in genital sexual arousal (80). Also, it has been shown that the distal vagina expresses arginase activity that is inhibited by ABH and BEC. Intravenous administration of the arginase inhibitor ABH enhanced vaginal engorgement as assessed by increased oxyhemoglobin content using near-infrared spectroscopy (Fig. 3) (72,80). The expression of arginase in the clitoris and vagina and the increase in blood ow observed after administration of the arginase inhibitor ABH suggest that this enzyme may play a role in modulating the NO pathway in female genital tissue and may regulate genital hemodynamics. Summary and conclusions The nitric oxide/cGMP pathway is deemed critical for male erectile function (1115,18). We and others have recently shown that the NO/cGMP pathway also plays a role in mod-

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dothelium in genital tissues may be reduced by increased arginase activity in response to disease or injury. Competition for the substrate L-arginine may be pronounced if L-arginine is present within a compartmentalized pool shared by these competing enzymes. Intracellular arginine availability may be further reduced in aging, diabetes, or atherosclerosis. If this were the case, it would be anticipated that alterations in arginase in various disease states would cause decreased NO production in response to sexual stimulation. These pathological changes would lead to insufcient perfusion or engorgement of genital tissues and dysfunctional sexual arousal responses in both men and women. As evidenced by the upregulation of arginase in specic disease states, its differential distribution in the vagina, and its modulation by sex steroid hormones, this enzyme may also participate in numerous other physiological and pathophysiological processes such as tissue growth, brosis, and immune function. LITERATURE CITED
1. NIH consensus conference (1993) Impotence. NIH consensus development panel impotence. J. Am. Med. Assoc. 270: 8390. 2. Laumann, E. O., Gagnon, J. H., Michael, R. T. & Michaels, S. (1994) The social organization of sexuality. University of Chicago Press, Chicago, IL. 3. Laumann, E. O., Paik, A. & Rosen, R. C. (1999) Sexual dysfunction in the United States: prevalence and predictors. J. Am. Med. Assoc. 281: 537544. 4. Feldman, H., Goldstein, I., Hatzichristou, D., Krane, R. & McKinlay, J. (1994) Impotence and its medical and psychosocial correlates: results of the Massachusetts male aging study. J. Urol. 151: 54 61. 5. Lue, T. F. & Tanagho, E. A. (1987) Physiology of erection and pharmacological management of impotence. J. Urol. 137: 829 836. 6. Krane, R. J., Goldstein, I. & Saenz de Tejada, I. (1989) Impotence. N. Engl. J. Med. 321: 1648 1659. 7. Saenz de Tejada, I., Moroukian, P., Tessier, J., Kim, J. J., Goldstein, I. & Frohrib, D. (1991) Trabecular smooth muscle modulates the capacitor function of the penis. Studies on a rabbit model. Am. J. Physiol. 260: H1590 H1595. 8. Andersson, K-E. & Wagner, G. (1995) Physiology of penile erection. Physiol. Rev. 75: 191236. 9. Giuliano, F., Allard, J., Compagnie, S., Alexandre, L., Droupy, S. & Bernabe, J. (2001) Vaginal physiological changes in a model of sexual arousal in anesthetized rats. Am. J. Physiol. 281: R140 R149. 10. Giuliano, F., Rampin, O. & Allard, J. (2002) Neurophysiology and pharmacology of female genital sexual response. J. Sex Marital. Ther. 28: 101 121. 11. Ignarro, L. J., Bush, P. A., Buga, G. M., Wood, K. S., Fukuto, J. M. & Rajfer, J. (1990) Nitric oxide and cyclic GMP formation upon electrical stimulation cause relaxation of corpus cavernosum smooth muscle. Biochem. Biophys. Res. Commun. 170: 843 850. 12. Kim, N., Azadzoi, K. M., Goldstein, I. & Saenz de Tejada, I. (1991) A nitric oxide-like factor mediates nonadrenergic-noncholinergic neurogenic relaxation of penile corpus cavernosum smooth muscle. J. Clin. Invest. 88: 112118. 13. Kim, N., Vardi, Y., Padma-Nathan, H., Daley, J., Goldstein, I. & Saenz de Tejada, I. (1993) Oxygen tension regulates the nitric oxide pathway. Physiological role in penile erection. J. Clin. Invest. 91: 437 442. 14. Rajfer, J., Aronson, W. J., Bush, P. A., Dorey, F. J. & Ignarro, L. (1992) Nitric oxide as a mediator of relaxation of the corpus cavernosum in response to nonadrenergic, noncholinergic neurotransmission. New Engl. J. Med. 326: 90 94. 15. Burnett, A. L., Lowenstein, C. J., Bredt, D. S., Chang, T. S. & Snyder, S. H. (1992) Nitric oxide: a physiologic mediator of penile erection. Science 17: 401 403. 16. Saenz de Tejada, I., Goldstein, I., Azadzoi, K., Krane, R. J. & Cohen, R. A. (1989) Impaired neurogenic and endothelium-mediated relaxation of penile smooth muscle from diabetic men with impotence. New Engl. J. Med. 320: 10251030. 17. Cartledge, J., Minhas, S. & Eardley, I. (2001) The role of nitric oxide in penile erection. Expert Opin. Pharmacother. 2: 95107. 18. Bolotina, V. M., Najibi, S., Palacino, J. J., Pagano, P. J. & Cohen, R. A. (1994) Nitric oxide directly activates calcium-dependent potassium channels in vascular smooth muscle. Nature 368: 850 853. 19. Gupta, S., Moreland, R. B., Munarriz, R., Daley, J., Goldstein, I. & Saenz de Tejada, I. (1995) Possible role of Na-K-ATPase in the regulation of human corpus cavernosum smooth muscle contractility by nitric oxide. Br. J. Pharmacol. 116: 22012206. 20. Burnett, A. L. (1997) Nitric oxide in the penis: physiology and pathology. J. Urol. 157: 320 324. 21. Garban, H., Vernet, D., Freedman, A., Rajfer, J. & Gonzalez-Cadavid, N. (1995) Effect of aging on nitric oxide-mediated penile erection in rats. Am. J. Physiol. 268: H467 475. 22. Dahiya, R., Lin, A., Bakircioglu, M. E., Huang, S. T. & Lue, T. F. (1997)

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FIGURE 3 Effects of ABH administration on vaginal and clitoral engorgement: Genital engorgement in response to pelvic nerve stimulation was assessed by near-infrared optical spectroscopy in anesthetized female New Zealand White rabbits. The pelvic nerve was electrically stimulated 10 min after intravenous injection of vehicle (control; 40% propylene glycol) or 4 6 mg/kg of ABH. Representative recordings are shown in the top panel. Amplitude, duration and area-underthe-curve (AUC) were determined for each response and the means SEM are shown for the relevant parameters in the 2 lower graphs (n 8; *P 0.05 compared to control). Reprinted with permission from Cama et al. (72), Copyright 2003, American Chemical Society.

ulating vaginal and clitoral hemodynamics (77,78,80,85). Alterations in the NO/cGMP pathway associated with aging, atherosclerosis, diabetes, hypercholesterolemia, or genital injury or trauma are likely to result in sexual dysfunction in men and women. The pathophysiological mechanisms of sexual dysfunction in male and female genitalia may involve: a) reduced NOS expression and activity, b) increased NO scavenging, c) increased synthesis of endogenous NOS inhibitors, or d) decreased availability of L-arginine as a substrate for neural and endothelial nitric oxide synthase. To date, very limited studies are available on the role of arginase activity in modulating the male and female genital arousal response (26,27,65,66,71,72). In this review, we summarized the data generated from our laboratory and those reported in the literature with regard to the potential regulatory role of arginase on erectile function and vaginal engorgement in vitro and in vivo. The availability of L-arginine as a substrate for nitric oxide synthase in NANC nerves and en-

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