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Unlike male mammals showing a well-delimited external urethral sphincter, female mammals have urogenital sphincters shaped by muscles like the urethrovaginal sphincter. Childbirth-related injuries affect morphometry and function of urogenital sphincters in women, which frequently underlies pelvic floor disorders, including stress urinary incontinence and pelvic organ prolapse. The bulboglandularis muscle (Bgm) seems to shape a urogenital sphincter in rabbits. We determined herein the effect of multiparity on urethral and vaginal pressures generated by the Bgm stimulation in age-matched nulliparous and multiparous chinchilla-breed rabbits to stimulate the Bgm with trains of ascending frequencies (from 1 to 100 Hz; 4 s duration each). Subsequently, the Bgm was excised, measured in width, and weighed. Significant differences (P ≤ 0.05) were determined with Mann-Whitney U or Student t-tests or repeated measures two-way ANOVA followed by Tukey tests. Spearman's partial coefficients were calculated to investigate the correlation between the highest pressure (urethral or vaginal) and the Bgm width. Multiparity reduced the weight and the width in the Bgm origin and medial regions. Urethral and vaginal pressures increased in response to the electrical stimulation of Bgm with frequencies from 20 to 100 Hz. Multiparas showed significant reductions in both types of pressures. We detected a strong correlation (conditioned by multiparity) between the medial Bgm width and the highest vaginal pressure. Our present findings demonstrate that multiparity impairs the function of Bgm, resulting in diminished urethral and vaginal pressures. Furthermore, the significant narrowness of the Bgm was correlated with the vaginal pressure recorded., (© 2023. The Author(s).)

Aim: To determine the effect of the lumbosacral ventral root avulsion (VRA) on the reflex activation of bladder, urethra, and activation of perineal muscles during micturition in female rabbits., Methods: We allocated 14 virgin female rabbits to evaluate, first, the gross anatomy of lumbosacral spinal cord root (n = 5) and, second, to determine the effect of VRA on perineal muscles during micturition (n = 9). We recorded cystometrograms, urethral pressure, and electromyograms of the bulbospongiosus (Bsm) and ischiocavernosus (Ism) muscles before and after the L6-S2 VRA. Standard variables were measured from each recording and analyzed to identify significant differences (P < .05)., Results: We found that the L6-S2 VRA affected directly the bladder and urethral function and reduced the duration and the frequency of the bursting of Ism and Bsm muscles during voiding. The Ism and Bsm showed a phasic activation, of different frequencies, during the voiding phase and the L6-S2 VRA inhibited the co-contraction of the Ism and Bsm-bladder-urethra., Conclusions: The Ism and Bsm are activated at different frequencies to trigger the voiding phase. The L6-S2 VRA affected the activity pattern of both perineal muscles. These modifications affected the bladder and urethra function. It is possible that the restoration of the activation frequency of perineal muscles contributed for an efficient bladder contraction., (© 2020 Wiley Periodicals, Inc.)

Aims: To evaluate the combined effect of age and multiparity on the micturition reflex, including pelvic floor muscle activation., Methods: Young and mature nulliparous rabbits were compared to young and mature multiparas (n = 6 per group). Cystometrograms and urethral pressure (UP) were performed while simultaneously recording the electromyogram (EMG) activity of the pubococcygeus and bulbospongiosus muscles to establish their functional correlation to urological function., Results: Multiparity and age significantly influence the bladder and UP affecting the voiding efficiency and intercontraction interval. Such interaction also reduced the UP threshold, timing, and duration. Other bladder and urethral variables were predominantly affected only by age. Urodynamic alterations correlated with abnormal patterns or absent EMG activity of the pubococcygeus and bulbospongiosus muscles., Conclusions: The present findings strongly suggest that multiparity and age affects specific pelvic floor muscle reflex activation during micturition, and may contribute to alterations in bladder and urethral function. This data broadens our understanding of the critical role of the appropriate activity of the individual pelvic floor muscles in micturition., (© 2019 Wiley Periodicals, Inc.)

Aim: To determine the anatomical organization and somatic axonal components of the lumbosacral nerves in female rabbits., Methods: Chinchilla adult anesthetized female rabbits were used. Anatomical, electrophysiological, and histological studies were performed., Results: L7, S1, and some fibers from S2 and S3 form the lumbosacral trunk, which gives origin to the sciatic nerve and innervation to the gluteal region. From S2 to S3 originates the pudendal nerve, whose branches innervates the striated anal and urethra sphincters, as well as the bulbospongiosus, ischiocavernosus, and constrictor vulvae muscles. The sensory field of the pudendal nerve is ∼1800 mm 2 and is localized in the clitoral sheath and perineal and perigenital skin. The organization of the pudendal nerve varies between individuals, three patterns were identified, and one of them was present in 50% of the animals. From S3 emerge the pelvic nerve, which anastomoses to form a plexus localized between the vagina and the rectum. The innervation of the pelvic floor originates from S3 to S4 fibers., Conclusions: Most of the sacral spinal nerves of rabbit are mixed, carrying sensory, and motor information. Sacral nerves innervate the hind limbs, pelvic viscera, clitoris, perineal muscles, inguinal and anal glands and perineal, perigenital, and rump skin. The detailed description of the sacral nerves organization, topography, and axonal components further the knowledge of the innervation in pelvic and perinal structures of the female rabbit. This information will be useful in future studies about the physiology and physiopathology of urinary, fecal, reproductive, and sexual functions., (© 2017 Wiley Periodicals, Inc.)

Aim: To determine the response of the pubococcygeus muscle (Pcm) during the urethrogenital reflex (UGR)., Methods: Urethane-anesthetized male rats (n = 20) were used to describe the gross anatomy of the Pcm (n = 3), the effect of the electrical stimulation of the Pcm on the urethral pressure (n = 3), and the reflex activity of the Pcm due to the mechanical stimulation of the urethra (n = 3) and during penile and urethrogenital reflexes (n = 11). The urethral pressure (UP) was recorded as a response to penile stimulation (brushing, extension, and occlusion) and during the UGR; the electromyographic activity of the Pcm was simultaneously evaluated. The role of the Pcm was assessed by measuring urethral pressure variables before and after denervation of this muscle., Results: The Pcm is innervated by the caudal branch of the somatomotor branch of the pelvic nerve. The electrically induced contraction of the Pcm increased the UP. The mechanical stimulation of the urethra during the induced micturition caused the reflex activity of the Pcm. The different penile stimuli caused bursts of activity of the Pcm. During the UGR, the Pcm exhibited a tonic activity. The transection of the caudal branch of the Smb reduced the maximal UP during the penile stimulation. The same was true regarding the duration of the UGR, the pressure that triggers the UGR, and the highest pressure observed during the UGR., Conclusion: Our results suggest that the activation of the Pcm is relevant for the UGR in male rats. Neurourol. Urodynam. 36:80-85, 2017. © 2015 Wiley Periodicals, Inc., (© 2015 Wiley Periodicals, Inc.)

Aim: To evaluate the morphometry and thyroid-hormone receptor (TR) expression in pelvic (pubococcygeus, Pcm) and perineal (bulbospongiosus, Bsm) muscles of control and hypothyroid female rabbits., Methods: Hypothyroidism was induced administering 0.02% methimazole in the drinking water for one month. Hematoxylin-eosin stained muscle sections were used to evaluate the fiber cross-sectional area (CSA) and the number of peripheral myonuclei per fiber. Immunohistochemistry was used to calculate the proportion of TR immunoreactive nuclei per fiber. Significant differences were considered at a P ≤ 0.05., Results: As compared to control rabbits, hypothyroidism increased the averaged fiber CSA and the myonuclei per fiber in the Bsm. Although the myonuclei number per fiber was also increased in the Pcm, the effect concerning the fiber CSA was only observed in a fraction of the Pcm fibers. Both TRα and TRβ were similarly expressed in the Pcm and Bsm. Hypothyroidism increased the expression of the TRα in the Bsm. Meanwhile, the expression of TR isoforms in the Pcm was not altered., Conclusion: Our findings support that the TR signaling is directly involved in morphometrical changes induced by hypothyroidism in the Pcm and Bsm. The effect of hypothyroidism on the Pcm and Bsm could be related to the different type of fiber and metabolism that these muscles have. Neurourol. Urodynam. 35:895-901, 2016. © 2015 Wiley Periodicals, Inc., (© 2015 Wiley Periodicals, Inc.)

Beyond its role in the conversion of androgens to estrogens, the expression of aromatase could influence on the estrogenic signalling in targeted tissues. Considering the well-defined biochemical and physiological differences between the pubococcygeus (Pcm) and bulbospongiosus (Bsm) muscles in female rabbits, it is presently hypothesized that the aromatase expression is differentially linked to the estrogen sensitivity of each muscle. To this end, serum estradiol levels and the aromatase expression, presence of ERα and ERβ and morphometry were evaluated in the Pcm and Bsm of female rabbits allocated in control, ovariectomized (OVX) and OVX treated with estradiol benzoate (OVX + EB) groups. Aromatase expression was high in the Pcm. Independently to serum estradiol, ovariectomy increased aromatase expression in the Pcm while decreased it in the Bsm. The EB treatment avoided the effect of ovariectomy only in the Pcm. The number of immunoreactive nuclei anti-ERα and anti-ERβ was high in the Pcm of OVX and OVX + EB rabbits, while those in the Bsm remained unchanged. The number of peripheral nuclei per fibre and the cross-sectional area-to-myonucleus ratio were modified only in the Pcm. Our findings support aromatase expression in the Pcm, and Bsm of rabbits is differentially linked to estrogenic sensitivity of each muscle., (Copyright © 2015 John Wiley & Sons, Ltd.)

In rodents, vaginal distention after delivery or experimental manipulation affects innervations as well as the amount of striated/smooth musculature or collagen in both the urethra and vagina. These changes are associated with modifications in excretory and reproductive processes. Although successive and consecutive vaginal deliveries (multiparity) affect the contractile and functional properties of the female lower urogenital tract (LUT), its impact on LUT morphometry, including persistency, has been barely studied. The caudal urethra (CU) and cranial (V1) and caudal (V2) pelvic vaginal regions were excised from young and adult nulliparous (YN and AN, respectively) and multiparous (YM and AM, respectively) rabbits. Tissues were histologically processed and stained with Masson's trichrome. The thickness of the tissue layers and areas covered by tissue components were measured and compared using two-way ANOVA followed by Student-Newmann-Keuls tests to determine statistical differences (P ≤ 0.05). Compared to YN, YM, and AN tissues showed a reduction in the thickness of the epithelium, as well as in areas covered by striated musculature, collagen, and blood vessels of the LUT. In comparison with YM, only some morphometric changes were recovered in the AM group. Our study shows that multiparity and age can be associated with epithelial and muscular atrophy of urethral and vaginal walls. The morphometry of the LUT between young and adult female rabbits varies with multiparity. These findings may help to better understand the effects of multiparity on young and adult females and its correlation with the development of pelvic dysfunctions., (© 2014 Wiley Periodicals, Inc.)

Aim: To evaluate the role of pelvic and perineal striated muscles on urethral function during micturition., Methods: Pubococcygeus, or both bulbospongiosus and ischiocavernosus muscles were electrically stimulated during the voiding phase of micturition, and bladder and urethral pressure were simultaneously recorded in urethane anesthetized female rabbits. Bladder and urethral function were assessed measuring urodynamic and urethral pressure variables obtained before and during the stimulation of muscles. Two-tail paired t-tests were carried out in order to determine significant differences (P < 0.05) between groups., Results: Electric stimulation of the pubococcygeus during voiding decreased voiding efficiency and voided volume, whereas residual volume, the duration of voiding, the interval between bladder contraction and urethral resistance increased. Simultaneously, there was an increase in maximum urethral pressure, as well as an increase in the pressure to return to baseline and in the pressure required to close the urethra. Electrical stimulation of bulbospongiosus and ischiocavernosus muscles increased voiding efficiency, voiding duration, and the maximal pressure in bladder. Meanwhile, the maximal urethral pressure, the time related to the rise of urethral pressure, and the urethral pressure required to close the urethra decreased., Conclusions: The stimulation of pelvic and perineal muscles have opposing roles in urethral function during micturition. Pubococcygeus muscles facilitate urethral closure, while they inhibit bladder contraction. In contrast, bulbospongiosus and ischiocavernosus muscles prevent urethral contraction while they promote bladder contraction., (© 2013 Wiley Periodicals, Inc.)

In European rabbits, the distal urethra (DU) opens into the anterior pelvic vagina forming a single canal by which females copulate, give birth, and urinate. We investigated whether the histological characteristics of the DU and the pelvic and perineal vagina are different. The DU and vagina of rabbits (Oryctolagus cuniculus) were cut and stained with hematoxylin-eosin and Masson's trichrome (n = 3). Data were compared by using Friedman's ANOVA for repeated measures. The walls of the DU and vagina are composed of mucosa, submucosa, smooth muscle, and an external layer. Differences in tissue characteristics of the mucosa, orientation of the smooth muscle fibers, components of the external layer (connective tissue, blood vessels, and striated musculature), and thickness of the tissue layers were found among regions. The lack of histological homogeneity along the urethra and vagina possibly reflects differences in the functions of each segment.