Tetsuji Soda

Authentic role of ATP signaling in micturition reflex.

Adenosine triphosphate (ATP) is a signaling molecule that regulates cellular processes. Based on previous studies of bladder function over the past decade, bladder ATP signaling was thought to have an essential role in the normal micturition reflex. In this study, we performed detailed analyses of bladder function in purinergic receptor-deficient mice using the automated voided stain on paper method and video-urodynamics. Unexpectedly, a lack of P2X2 or P2X3 receptors did not affect bladder function under normal physiological conditions, indicating that bladder ATP signaling is not essential for normal micturition reflex. In contrast, we found that lipopolysaccharide (LPS) induced markedly high levels of ATP release from the urothelium. In addition, LPS-induced rapid bladder hyperactivity was attenuated in P2X2−/− and P2X3−/− mice. Contrary to the previous interpretation, our present findings indicate that bladder ATP signaling has a fundamental role in the micturition reflex, especially in bladder dysfunction, under pathological conditions. Therefore, the bladder ATP signaling pathway might be a highly promising therapeutic target for functional bladder disorders. This study newly defines an authentic role for bladder ATP signaling in the micturition reflex.

Sexual life of Japanese patients with erectile dysfunction taking phosphodiesterase type 5 inhibitors: An Internet survey using the Psychological and Interpersonal Relationship Scales-Short Form questionnaire

To investigate details of sexual function of erectile dysfunction in Japanese patients taking phosphodiesterase type 5 inhibitors.

Combination of bladder ultrasonography and novel cystometry method in mice reveals rapid decrease in bladder capacity and compliance in LPS-induced cystitis

Various animal models have been used in research into bladder dysfunction, and in vivo cystometry is a common method to analyze bladder function in animals. However, it is rather difficult to perform reliably in small animals. Transabdominal bladder ultrasonography combined with cystometry in urethane-anesthetized mice have revealed physical inhibition of bladder wall movement by a bladder catheter conventionally placed in the bladder apex. For reliable evaluation of mouse lower urinary tract function, we established a novel cystometry method in which a catheter was placed in the bladder anterior wall, in combination with bladder ultrasonography. This new method allowed the bladder to be well distended (i.e., larger maximum bladder capacity, lower pressure threshold, higher voided volume, and higher bladder compliance compared with conventional methods), which reflected more spontaneous voiding than conventional cystometry methods. We also demonstrated the usefulness of bladder ultrasonography for analysis of mouse bladder function, especially bladder dynamics, maximum bladder capacity, and post-voiding residual volume. We analyzed bladder functional changes in lipopolysaccharide (LPS)-induced cystitis by combining bladder ultrasonography and this new cystometry method. Bladder ultrasonography revealed a rapid decrease in bladder capacity, and cystometry showed a rapid decrease in voided volume due to intravesical LPS instillation. This new cystometry method also revealed a rapid decrease in bladder compliance caused by LPS instillation, which was not detectable by conventional methods. The combination of ultrasonography and the new cystometry method may become a powerful tool for analysis of mouse bladder function and could contribute to the development of new treatments for bladder dysfunction.

Histologic Evaluation of Human Benign Prostatic Hyperplasia Treated by Dutasteride: A Study by Xenograft Model With Improved Severe Combined Immunodeficient Mice

OBJECTIVE To evaluate histologic change in human prostate samples treated with dutasteride and to elucidate direct effects of dutasteride on human prostate tissue, the present study was conducted by using a xenograft model with improved severe combined immunodeficient (super-SCID) mice, although it is well known that dutasteride reduces prostate volume. METHODS After establishment of a xenograft model of human benign prostatic hyperplasia in morphology and function, samples implanted into super-SCID mice with and without dutasteride were evaluated pathohistologically at 2 and 6 months after initiation of dutasteride administration. RESULTS The proliferative index evaluated by Ki-67 staining was significantly lower in the dutasteride group than the control at 2 and 6 months after administration. Apoptotic index evaluated by the terminal transferase TdT-mediated dUTP-biotin nick end labeling staining was higher in the dutasteride group than the control at 2 and 6 months after administration. Quick scores in the dutasteride group for staining of both cyclooxygenase-2 (Cox-2) and Ras homolog gene family, member A (RhoA) were significantly lower than those in the control group at 2 and 6 months after administration. CONCLUSION Dutasteride inhibits cell proliferation and induces apoptosis of prostatic cells, causing a reduced prostate volume. Furthermore, decreased expression of Cox-2 and RhoA within benign prostatic hyperplasia tissue by dutasteride may induce an early effect on improvement of lower urinary tract symptoms, probably by attenuating inflammation reaction of the prostate and decreasing intraurethral pressure, other than the mechanism of reduced prostate volume.

Systematic characterization of human testis-specific actin capping protein β3 as a possible biomarker for male infertility

STUDY QUESTION Is actin capping protein (CP) β3 involved in human spermatogenesis and male infertility? SUMMARY ANSWER Human CPβ3 (hCPβ3) is expressed in testis, changes its localization dynamically during spermatogenesis, and has some association with male infertility. WHAT IS KNOWN ALREADY The testis-specific α subunit of CP (CPα3) was previously identified in human, and mutations in the cpα3 gene in mouse were shown to induce malformation of the sperm head and male infertility. However, CPβ3, which is considered to be a heterodimeric counterpart of CPα3, has been neither characterized in human nor reported in association with male infertility. STUDY DESIGN, SIZE, DURATION To confirm the existence of CPβ3 in human testis, fresh semen samples from proven fertile men were analyzed. To investigate protein expression during spermatogenesis, cryopreserved testis obtained from men with obstructive azoospermia were examined by immunofluorescent analysis. To assess the association of CP with male infertility, we compared protein expression of human CPα3 (hCPα3) and hCPβ3 using immunofluorescent analysis of cryopreserved sperm between men with normozoospermia (volunteers: Normo group, n = 20) and infertile men with oligozoospermia and/or asthenozoospermia (O + A group, n = 21). PARTICIPANTS/MATERIALS, SETTING, METHODS The tissue-specific expression of hCPβ3 was investigated by RT-PCR and Western blot analysis. To investigate whether hCPα3 and hCPβ3 form a heterodimer, a tandem expression vector containing hcpα3 tagged with monomeric red fluorescent protein 1 and hcpβ3 tagged with enhanced green fluorescent protein in a single plasmid was constructed and analyzed by co-immunoprecipitation (Co-IP) assay. The protein expression profiles of hCPα3 and hCPβ3 during spermatogenesis were examined by immunohistochemical analysis using human spermatogenic cells. The protein expressions of hCPα3 and hCPβ3 in sperm were compared between the Normo and O + A groups by immunohistochemical analysis. MAIN RESULTS AND THE ROLE OF CHANCE RT-PCR showed that mRNA of hcpβ3 was expressed exclusively in testis. Western blot analysis detected hCPβ3 with anti-bovine CPβ3 antibody. Co-IP assay with recombinant protein showed that hCPα3 and hCPβ3 form a protein complex. At each step during spermatogenesis, the cellular localization of hCPβ3 changed dynamically. In spermatogonia, hCPβ3 showed a slight signal in cytoplasm. hCPβ3 expression was conspicuous mainly from spermatocytes, and hCPβ3 localization dynamically migrated from cytoplasm to the acrosomal cap and acrosome. In mature spermatozoa, hCPβ3 accumulated in the postacrosomal region and less so at the midpiece of the tail. Double-staining analysis revealed that hCPα3 localization was identical to hCPβ3 at every step in the spermatogenic cells. Most spermatozoa from the Normo group were stained homogenously by both hCPα3 and hCPβ3. In contrast, significantly more spermatozoa in the O + A versus Normo group showed heterogeneous or lack of staining for either hCPα3 or hCPβ3 (abnormal staining) (P < 0.001). The percentage of abnormal staining was higher in the O + A group (52.4 ± 3.0%) than in the Normo group (31.2 ± 2.5%). Even by confining the observations to morphologically normal spermatozoa selected in accordance with Davids criteria, the percentage of abnormal staining was still higher in the O + A group (39.9 ± 2.9%) versus the Normo group (22.5 ± 2.1%) (P < 0.001). hCPβ3 in conjunction with hCPα3 seemed to play an important role in spermatogenesis and may be associated with male infertility. LARGE SCALE DATA Not applicable. LIMITATIONS REASONS FOR CAUTION Owing to the difficulty of collecting fresh samples of human testis, we used cryopreserved samples from testicular sperm extraction. To examine the interaction of spermatogenic cells or localization in seminiferous tubules, fresh testis sample of healthy males are ideal. WIDER IMPLICATIONS OF THE FINDINGS The altered expression of hCPα3 and hCPβ3 may not only be a cause of male infertility but also a prognostic factor for the results of ART. They may be useful biomarkers to determine the fertilization ability of human sperm in ART. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by a Grant-in-Aid for Young Scientists (B) from the Japan Society for the Promotion of Science (JP16K20133). The authors declare no competing interests.

Intravesical ATP instillation induces urinary frequency because of activation of bladder afferent nerves without inflammatory changes in mice: A promising model for overactive bladder

ATP in the suburothelial layer is released from the bladder urothelium by mechanical stimuli. ATP directly activates purinergic receptors that are expressed on primary bladder afferent neurons and induces the micturition reflex. Although ATP is also released to the bladder lumen from the bladder urothelium, the role of ATP in the bladder lumen is unknown. Recently, clinical studies have reported that urinary ATP levels are much higher in patients with an overactive bladder than healthy controls. These results suggest that ATP in the bladder lumen is also involved in the micturition reflex. In this study, we performed intravesical ATP instillation in the mouse bladder. We evaluated urinary function with novel reliable methods using improved cystometry and ultrasonography, which we previously established. We found that intravesical ATP instillation induced urinary frequency because of activation of bladder afferent nerves without inflammatory changes in the bladder or an increase in post-void residual urine. These results suggest that not only ATP in the suburothelial layer, but also ATP in the bladder lumen, are involved in enhancement of the micturition reflex.

PD70-06 THE ROLE OF UROTHELIAL ATP SIGNALING IN MICTURITION REFLEX

nociceptive neurons (TRPV1-ChR2 and Nav 1.8-Arch). These opsins were activated be either blue (ChR2) or green (Arch) laser or LED illumination of the bladder. We then measured the influence of opsin activation on bladder function using continuous infusion cystometry. In order to translate this into a more clinically relevant gene delivery system, bladder wall injection of herpes simplex viral vector (HSV) with a pan-neuronal promotor controlling expression of ChR2 or Arch were used in rats. RESULTS: In TRPV1-ChR2 mice, ChR2 activation with blue light resulted in voiding events in a half full bladder that increased in magnitude with increase of light intensity. Activation of Arch with green light could delay regular cystometric contractions in Nav 1.8-Arch mice. In rats transduced with HSV-ChR2, we were able to evoke voiding events with ChR2 activation by blue light illumination of the bladder, after less filling than under normal conditions (non-stimulated). These effects were not seen in rats injected with the control viral vector, HSVeYFP. Finally, illumination of the bladder with green light delayed regular cystometric contractions in rats injected with HSV-Arch but not in animals injecting with HSV-eYFP. CONCLUSIONS: Here we demonstrate bidirectional modulation of bladder function using optogenetics.. This bidirectional control of bladder function can be induced by selective targeting of nociceptive afferents. Further restriction of opsin expression to specific populations of bladder sensory fibers could lead to a better understanding of their role in bladder function and disease. The refinement of virally delivery methods for optogenetic proteins, together with the development of medical devices for light delivery could lead to development of future therapies for bladder dysfunction.

MP84-14 THE RELATIONSHIPS BETWEEN LIFESTYLE AND TESTOSTERONE; NIGHT WAKING, SMOKING HISTORY, DRINK HABIT AND THE LENGTH OF TIME USING INTERNET AND PLAYING GAMES ARE ASSOCIATED WITH TESTOSTERONE LEVEL

CONCLUSIONS: Only CFT and T:E ratio were predictive of positive libido response on IIEF11 & 12 questionnaire in our cohort. Estradiol, even at a cutoff of 5 ng/dL, was not independently associated with improved libido. Surprisingly total testosterone did not associate with IIEF11 (desire frequency). The effect of testosterone and estradiol administration on libido requires further prospective study.

The Pattern of Sexual Interest of Female-to-Male Transsexual Persons With Gender Identity Disorder Does Not Resemble That of Biological Men: An Eye-Tracking Study

Introduction Very little has been elucidated about sexual interest in female-to-male (FtM) transsexual persons. Aims To investigate the sexual interest of FtM transsexual persons vs that of men using an eye-tracking system. Methods The study included 15 men and 13 FtM transsexual subjects who viewed three sexual videos (clip 1: sexy clothed young woman kissing the region of the male genitals covered by underwear; clip 2: naked actor and actress kissing and touching each other; and clip 3: heterosexual intercourse between a naked actor and actress) in which several regions were designated for eye-gaze analysis in each frame. The designation of each region was not visible to the participants. Main Outcome Measures Visual attention was measured across each designated region according to gaze duration. Results For clip 1, there was a statistically significant sex difference in the viewing pattern between men and FtM transsexual subjects. Longest gaze time was for the eyes of the actress in men, whereas it was for non-human regions in FtM transsexual subjects. For clip 2, there also was a statistically significant sex difference. Longest gaze time was for the face of the actress in men, whereas it was for non-human regions in FtM transsexual subjects, and there was a significant difference between regions with longest gaze time. The most apparent difference was in the gaze time for the body of the actor: the percentage of time spent gazing at the body of the actor was 8.35% in FtM transsexual subjects, whereas it was only 0.03% in men. For clip 3, there were no statistically significant differences in viewing patterns between men and FtM transsexual subjects, although longest gaze time was for the face of the actress in men, whereas it was for non-human regions in FtM transsexual subjects. Conclusion We suggest that the characteristics of sexual interest of FtM transsexual persons are not the same as those of biological men. Tsujimura A, Kiuchi H, Soda T, et al. The Pattern of Sexual Interest of Female-to-Male Transsexual Persons With Gender Identity Disorder Does Not Resemble That of Biological Men: An Eye-Tracking Study. Sex Med 2017;5:e169–e174.

MP71-16 GENERATION DIFFERENCE ON THE ASSOCIATION BETWEEN LOWER URINARY TRACT SYMPTOMS AND LATE-ONSET HYPOGONADISM SYMPTOMS: ANALYSIS OF 1398 HEALTHY YOUNG AND ELDERLY ADULTS

INTRODUCTION AND OBJECTIVES: Aging is known to affect sexual, psychological and physiological functions including lower urinary tract symptoms (LUTS). Recent data suggested that severity of LUTS was associated with that of late-onset hypogonadism (LOH) in elderly men. However, information on the association in young adults is limited. In this study, we evaluated the relationship between LUTS and LOH symptoms in young and elderly adults. METHODS: This study included 1398 healthy men 20 years; 130 in 20,fs, 519 in 30,fs, 399 in 40,fs, 268 in 50,fs and 82 in 60,fs. LUTS was assessed using I-PSS questionnaire and LOH symptoms was evaluated using Aging Male’s Symptoms (AMS) questionnaire with higher score indicating more severe symptoms. LUTS was diagnosed as I-PSSS 8. Severity of LUTS was defined by I-PSS-QOL as follows; 0-1 as mild, 2-3 as moderate, and 4-6 as severe. Generalized additive models were used for statistical analysis. RESULTS: Total I-PSS score increased with age: LUTS was observed in 3.1% in 20’s, 2.7% in 30’s, 7.8% in 40’s, 15.3% in 50’s and 22.0% in 60’s (p for trend <0.01) (Fig.1). Mild LUTS was found in 751(54%), and moderate LUTS in 556(40%), severe LUTS in 91(7%). Severity of LOH symptoms increased with increasing LUT severity in any years of age. Moreover, young adults with severe LUTS had more severe LOH symptoms than elderly adults (36.3 vs. 28.5, p<0.05) (Fig. 2); they reported higher psychological and physiological sub-score than elderly men, whereas they reported lower sexual score. CONCLUSIONS: Both young and elderly adults with severe LUTS have severe LOH symptoms, but young adults with severe LUTS were more likely to have severe LOH symptoms than elderly adults. Therefore, urologist should take care of hypogonadism symptoms other than LUTS when young adults are presented with LUTS. Source of Funding: none