Shift from Pro- to Anti-Inflammatory Phase in Pelvic Floor Muscles at Postpartum Matches Histological Signs of Regeneration in Multiparous Rabbits.

Background and Objectives: Pelvic floor muscles (PFM) play a core role in defecation and micturition. Weakening of PFM underlies urogynecological disorders such as pelvic organ prolapse and stress urinary incontinence. Vaginal delivery damages PFM. Muscle trauma implies an inflammatory response mediated by myeloid cells, essential for subsequent recovery. Molecular signaling characterizing the pro-inflammatory phase shifts M1 macrophages to M2 macrophages, which modulate muscle repair. The present study aimed to evaluate histological characteristics and the presence of M1 and M2 macrophages in bulbospongiosus (Bsm) and pubococcygeus muscles (Pcm). Materials and Methods: Muscles from young nulliparous (N) and multiparous rabbits on postpartum days three (M3) and twenty (M20) were excised and histologically processed to measure the myofiber cross-sectional area (CSA) and count the centralized myonuclei in hematoxylin-eosinstained sections. Using immunohistochemistry, M1 and M2 macrophages were estimated in muscle sections. Kruskal-Wallis or one-way ANOVA testing, followed by post hoc tests, were conducted to identify significant differences (p < 0.05). Results: The myofiber CSA of both the Bsm and Pcm of the M3 group were more extensive than those of the N and M20 groups. Centralized myonuclei estimated in sections from both muscles of M20 rabbits were higher than those of N rabbits. Such histological outcomes matched significant increases in HLA-DR immunostaining in M3 rabbits with the CD206 immunostaining in muscle sections from M20 rabbits. Conclusions: A shift from the pro- to anti-inflammatory phase in the bulbospongiosus and pubococcygeus muscles of multiparous rabbits matches with centralized myonuclei, suggesting the ongoing regeneration of muscles.

Sucrose consumption modifies the urethrogenital reflex and histological organization of the bulbospongiosus muscle in the male rat.

Disorders of the bulbospongiosus muscle (Bsm) are associated with male sexual dysfunction, such as premature ejaculation. We determined the effect of sucrose-water consumption during pregnancy-lactation and postnatal on reflex responses and morphology of Bsm fibers in adult male Wistar rat offspring. Female rats were mated and grouped into consumed tap water mothers and sucrose-water (5 %) mothers during pregnancy-lactation to obtain experimental groups. Male pups were weaned and assigned into four groups (n = 12; each group). Those from control mothers who continued drinking tap water (CM-CO group) or sucrose water (CM-SO group), and those from sucrose mothers who drank tap water (SM-CO group) or continued drinking sucrose water (SM-SO group) until adult life. In male rat offspring (n = 6 per group) was recorded the electrical activity of Bsm was recorded during penile stimulation and urethrogenital reflex (UGR). Other male rat offspring were designated for histological analysis (n = 6 per group). Sucrose consumption during prenatal stages increased the frequency of the Bsm during UGR, while pre and postnatal consumption modified muscle fiber cross-sectional area and increased the collagen content, suggesting that a combination of a diet with pre- and postnatal sucrose changes the Bsm morphophysiology possibly causing male sexual dysfunctions.

Preventive Effect of Molecular Iodine in Pancreatic Disorders from Hypothyroid Rabbits.

Pancreatic alterations such as inflammation and insulin resistance accompany hypothyroidism. Molecular iodine (I2) exerts antioxidant and differentiation actions in several tissues, and the pancreas is an iodine-uptake tissue. We analyzed the effect of two oral I2 doses on pancreatic disorders in a model of hypothyroidism for 30 days. Adult female rabbits were divided into the following groups: control, moderate oral dose of I2 (0.2 mg/kg, M-I2), high oral dose of I2 (2.0 mg/kg, H-I2), oral dose of methimazole (MMI; 10 mg/kg), MMI + M-I2,, and MMI + H-I2. Moderate or high I2 supplementation did not modify circulating metabolites or pancreatic morphology. The MMI group showed reductions of circulating thyroxine (T4) and triiodothyronine (T3), moderate glucose increments, and significant increases in cholesterol and low-density lipoproteins. Acinar fibrosis, high insulin content, lipoperoxidation, and overexpression of GLUT4 were observed in the pancreas of this group. M-I2 supplementation normalized the T4 and cholesterol, but T3 remained low. Pancreatic alterations were prevented, and nuclear factor erythroid-2-related factor-2 (Nrf2), antioxidant enzymes, and peroxisome proliferator-activated receptor gamma (PPARG) maintained their basal values. In MMI + H-I2, hypothyroidism was avoided, but pancreatic alterations and low PPARG expression remained. In conclusion, M-I2 supplementation reestablishes thyronine synthesis and diminishes pancreatic alterations, possibly related to Nrf2 and PPARG activation.

Multiparity Reduces Urethral and Vaginal Pressures Following the Bulboglandularis Muscle Stimulation in Rabbits.

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.

Secondary urethral sphincter function of the rabbit pelvic and perineal muscles.

Perineal and pelvic floor muscles play an important role in continence by providing mechanical support to pelvic organs. It is also known that the pubococcygeus muscle (PcM) contracts in the storage phase and is inactive during voiding, while the bulbospongiosus muscle (BsM) is active during the voiding phase. Recent evidence suggested an additional role of these muscles in supporting urethral closure in rabbits. However, the individual role of perineal and pelvic muscles as urethral sphincters is not well-defined. Here we evaluated the individual, sequential and synergistic roles of the PcM and BsM in assisting urethral closure and defined the optimal electrical stimulation parameters that can effectively contract these muscles and increase the urethral pressure (P ura ) in young nulliparous animals (n = 11). Unilateral stimulation of either the BsM or PcM at 40 Hz induced modest increases in average P ura (0.23 ± 0.10 and 0.07 ± 0.04 mmHg, respectively). Investigation on the changes in P ura evoked by stimulation frequencies between 5 and 60 Hz show that sequential contralateral PcM-BsM activation at 40 Hz induced a 2-fold average P ura increase (0.23 ± 0.07 mmHg) compared to that evoked by PcM stimulation. Simultaneous activation of PcM and BsM at 40 Hz also showed an increased average P ura (0.26 ± 0.04 mmHg), with a 2-fold increase in average P ura observed during the unilateral sequential PcM-BsM stimulation at 40 Hz (0.69 ± 0.2 mmHg). Finally, stimulation at 40 Hz of the bulbospongiosus nerve (BsN) induced an approximate 4-fold increase in average P ura (0.87 ± 0.44 mmHg; p < 0.04) compared to that elicited by BsM stimulation, confirming that direct nerve stimulation is more effective. Together, this study shows that in the female rabbit, both perineal and pelvic muscles support of the urethral function during continence, and that unilateral stimulation of the BsN at 40-60 Hz is sufficient to achieve maximal secondary sphincter activity. The results also support the potential clinical value of neuromodulation of pelvic and perineal nerves as bioelectronic therapy for stress urinary incontinence.

High-sucrose diet-fed male rats show affectations in pubococcygeus reflex activation and myofiber content.

In males, the function of the pubococcygeus muscle (Pcm) shows a high androgen dependency. High sucrose intake increases testosterone levels in pubertal male rats. We aimed to analyze the Pcm reflex activity and fiber type in response to a 30 % sucrose diet. Wistar male rats were assigned into two groups: control and sugar. The Pcm electromyographic activity was recorded during the mechanical stimulation of the scrotal skin and penis. We also determined the relative levels of Glut4 expression, glycogen content, myofiber cross-sectional area (CSA), and the content of glycolytic and oxidative with NADH-TR or fast, intermediate, and slow fibers with alkaline (pH 9.4) ATPase histochemistry. The sugar group showed a short Pcm reflex activity, a higher Glut4 expression, and glycogen content; the CSA fibers showed a significant difference in the percentage of fibers with the different transversal areas. The number of nuclei was positively correlated with the CSA of the Pcm fibers. There was also a decrease in oxidative fibers but an increase in glycolytic fibers, while the content of fast muscle fibers increased to the detriment of intermediate and slow fibers. Thus, a high-sugar diet reduced Pcm reflex activity, fiber type, and dysregulation of skeletal muscle energy metabolism.

G protein-coupled estrogen receptor (GPER/GPR30) levels in pelvic floor muscles and its association with estrogen status in female rabbits.

Objective: To assess the relative expression of the G-protein coupled estrogen receptor (GPER) in the bulbospongiosus (Bsm) and pubococcygeus (Pcm) muscles in control, ovariectomized (OVX), and OVX with estradiol benzoate supplementation (OVX + EB) rabbits.Methods: We used tissues from C, 1-month OVX, and OVX plus 15-day EB implanted (OVX + EB) groups. The GPER expression was evaluated by Western blot and immunohistochemistry for both Bsm and Pcm. Results: Both muscles showed a GPER immunoreactivity in blood vessels, inside myofibers next to myonuclei, and in polymorphonuclear cells. Four-week ovariectomy did not modify the GPER expression in the Bsm and Pcm, but two-week estradiol benzoate increased it in the latter muscle alone.Conclusions: We demonstrated that the Bsm and Pcm of female rabbits express GPER. High serum estradiol levels elevate GPER relative expression in the Pcm alone. The present study supports the remarkable estrogen sensitivity of the Pcm.

Effects of hypothyroidism on the female pancreas involve the regulation of estrogen receptors.

This study aimed to investigate the impact of short-time hypothyroidism on the expression of aromatase, estrogen receptors (ERα, ß), and GPR30 in the pancreas of female rabbits. The formation of new islets and the expression of insulin, GLUT4, and lactate dehydrogenase (LDH) were also analyzed. This purpose is based on actions that thyroid hormones and estrogens have on ß-cells differentiation, acinar cell function, and insulin secretion. Twelve Chinchilla-breed adult virgin female rabbits were divided into control (n = 6) and hypothyroid (n = 6; methimazole 10 mg/kg for 30 days) groups. In the complete pancreas, expressions of aromatase and estrogen receptors, as well as proinsulin, GLUT4, and LDH were determined by western blot. Characteristics of islets were measured in slices of the pancreas with immunohistochemistry for insulin. Islet and acinar cells express aromatase, ERα, ERß, and GPR30. Hypothyroidism increased the expression of ERα and diminished that for aromatase, ERß, and GPR30 in the pancreas. It also promoted a high number of extra small islets (new islets) and increased the expression of proinsulin and GLUT4 in the pancreas. Our results show that actions of thyroid hormones and estrogens on ß-cells neogenesis, acinar cell function, and synthesis and secretion of insulin are linked. Thus, the effects of hypothyroidism on the pancreas could include summatory actions of thyroid hormones plus estrogens. Our findings indicate the importance of monitoring estrogen levels and actions on the pancreas of hypothyroid women, particularly when serum estrogen concentrations are affected such as menopausal, pregnant, and those with contraceptive use.

Estrogens influence differentially on the pelvic floor muscles activation at somatovisceral reflexes involved in micturition of rabbits.

OBJECTIVE: To determine the estrogen-dependency of the bladder and urethral function and the coordinated activation of pelvic floor muscles (PFM) during micturition. METHODS: We allocated age-matched female rabbits to control, 1-month ovariectomized (OVX), and OVX plus 2-week estradiol benzoate (EB) groups to record cystometry, urethral pressure, and electromyograms of bulbospongiosus (Bsm), and pubococcygeus muscles (Pcm) simultaneously. We also measured serum estradiol levels and myofiber cross-sectional area. We assessed urodynamic and urethral variables, categorized the Bsm-Pcm activation patterns at storage and voiding phases, and obtained the power spectrum density of muscle activation around the voiding phase. We investigated the influence of ovarian hormones, in general, and the contribution of estrogen, particularly on the functions of the bladder, urethra, and PFM. Statistical significance was set at P < 0.05. RESULTS: Ovarian hormones influence the bladder, urethral, and PFM functions. The urodynamics analyses indicated estrogens contribute to voiding duration and, to a lesser extent, to the time between bladder contractions. Urethral pressure at closure (maximal pressure-to-maximal urethral pressure ratio) improved partially (8%, P < 0.05) in the OVX plus 2-week estradiol benzoate compared with OVX, but urethral resistance increased (∼1.9-fold, P < 0.05) compared with control rabbits. Our findings support that Pcm activity at voiding is estrogen-sensitive, albeit EB administration reduced it at storage resume, which relates to high urethral resistance. CONCLUSIONS: Ovariectomy impairs bladder and urethral pressures and Bsm and Pcm activation at micturition in anesthetized rabbits. Estrogen administration partially reverts some of these effects and influences Pcm activation.

Targeted neuromodulation of pelvic floor nerves in aging and multiparous rabbits improves continence.

Pelvic floor muscle stretch injury during pregnancy and birth is associated with the incidence of stress urinary incontinence (SUI), a condition that affects 30-60% of the female population and is characterized by involuntary urine leakage during physical activity, further exacerbated by aging. Aging and multiparous rabbits suffer pelvic nerve and muscle damage, resulting in alterations in pelvic floor muscular contraction and low urethral pressure, resembling SUI. However, the extent of nerve injury is not fully understood. Here, we used electron microscopy analysis of pelvic and perineal nerves in multiparous rabbits to describe the extent of stretch nerve injury based on axon count, axon size, myelin-to-axon ratio, and elliptical ratio. Compared to young nulliparous controls, mid-age multiparous animals showed an increase in the density of unmyelinated axons and in myelin thickness in both nerves, albeit more significant in the bulbospongiosus nerve. This revealed a partial but sustained damage to these nerves, and the presence of some regenerated axons. Additionally, we tested whether electrical stimulation of the bulbospongiosus nerve would induce muscle contraction and urethral closure. Using a miniature wireless stimulator implanted on this perineal nerve in young nulliparous and middle age multiparous female rabbits, we confirmed that these partially damaged nerves can be acutely depolarized, either at low (2-5 Hz) or medium (10-20 Hz) frequencies, to induce a proportional increase in urethral pressure. Evaluation of micturition volume in the mid-age multiparous animals after perineal nerve stimulation, effectively reversed a baseline deficit, increasing it 2-fold (p = 0.02). These results support the notion that selective neuromodulation of pelvic floor muscles might serve as a potential treatment for SUI.

Differential estrogen-related responses in myofiber cross-sectional area of pelvic floor muscles in female rats.

OBJECTIVE: To determine the role of estrogens in myofiber cross-sectional area (CSA) of the pubococcyegeus (Pcm) and iliococcygeus muscles (Icm). METHODS: In Experiment 1, we excised the Pcm and Icm during the metestrus and proestrus stages of the estrous cycle to measure the myofiber CSA. In Experiment 2, we allocated other rats into the following groups: sham (Sh), ovariectomized (OVX), OVX plus 1,4,6-androstatriene-3,17-dione (ATD; OVX + ATD), an aromatase inhibitor, and OVX plus estradiol benzoate (OVX + EB). We carried out appropriate statistical tests to determine significant differences (p ≤ 0.05) in variables measured for both Experiments. RESULTS: The Pcm myofiber CSA at proestrus was higher than at metestrus, while the Icm myofiber CSA did not change. Ovariectomy increased the Pcm myofiber CSA, which was exacerbated with the ATD administration. The EB supplementation successfully reversed the ovariectomy-induced enlargement of the CSA. No significant changes were detected for the Icm myofiber CSA. CONCLUSIONS: Fluctuating ovarian steroid levels at the estrus cycle significantly influence the CSA myofiber of the Pcm but not that of the Icm. Estrogen actions, having a gonadal or extragonadal origin, influence importantly the CSA of the Pcm.

Heat shock proteins and antioxidants as mechanisms of response to ivermectin in the dung beetle Euoniticellus intermedius.

Ivermectin is the most common antiparasitic drug used in livestock in many regions of the world. Its residues are excreted in dung, threatening non-target fauna such as dung beetles, fundamental for cleaning dung in pastures. However, it is unclear which are the physiological mechanisms used by dung beetles to cope with ivermectin. Here we evaluated experimentally the physiological responses of the dung beetle Euoniticellus intermedius to ivermectin-induced stress. We measured metabolic rates, heat shock protein 70 (Hsp70) expression, antioxidant capacity, and oxidative damage in lipids in both males and females exposed to a sublethal dose. Compared to control beetles, ivermectin-treated males and females had increased metabolic rates. Moreover, ivermectin-treated females increased their expression of Hsp70 whereas males increased their antioxidant capacity. No changes in the levels of oxidative damage to lipids were detected for either sex, suggesting a process of hormesis, such that exposure to a moderate concentration of ivermectin could stimulate the action of a protective mechanism against oxidative stress, that differs between sexes. However, it does not exclude the possibility that damage to other biomolecules might have occurred. Sexual differences in physiological responses can be interpreted as the result of hormonal differences or life-history trade-offs that favor different mechanisms in females and males. Hsps and antioxidants are involved in the physiological response of beetles to ivermectin and may be key in providing resistance to this contaminant in target and non-target species, including dung beetles.

Lumbosacral ventral root avulsion alters reflex activation of bladder, urethra, and perineal muscles during micturition in female rabbits.

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.

The Expression of Hormone Receptors as a Gateway toward Understanding Endocrine Actions in Female Pelvic Floor Muscles.

OBJECTIVE: To provide an overview of the hormone actions and receptors expressed in the female pelvic floor muscles, relevant for understanding the pelvic floor disorders. METHODS: We performed a literature review focused on the expression of hormone receptors mainly in the pelvic floor muscles of women and female rats and rabbits. RESULTS: The impairment of the pelvic floor muscles can lead to the onset of pelvic floor dysfunctions, including stress urinary incontinence in women. Hormone milieu is associated with the structure and function alterations of pelvic floor muscles, a notion supported by the fact that these muscles express different hormone receptors. Nuclear receptors, such as steroid receptors, are up till now the most investigated. The present review accounts for the limited studies conducted to elucidate the expression of hormone receptors in pelvic floor muscles in females. CONCLUSION: Hormone receptor expression is the cornerstone in some hormone-based therapies, which require further detailed studies on the distribution of receptors in particular pelvic floor muscles, as well as their association with muscle effectors, involved in the alterations relevant for understanding pelvic floor disorders.

Bladder and urethral dysfunction in multiparous and mature rabbits correlates with abnormal activity of pubococcygeus and bulbospongiosus muscles.

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.

Signs of damage in pelvic floor muscles at the end of pregnancy in rabbits.

INTRODUCTION AND HYPOTHESIS: Temporary effects to pelvic floor muscles are linked to impairments in micturition, particularly stress urinary incontinence (SUI), during pregnancy. We hypothesize that bulbospongiosus (Bsm) and pubococcygeus (Pcm) are differently damaged in primigravid and primiparous rabbits. METHODS: Twenty-four rabbits allocated evenly (n = 6) into nulliparous, pregnant, and primiparous groups on postpartum days 3 (P3) and 20 (P20) were used to evaluate the myofiber cross-sectional area (CSA), ß-glucuronidase activity, and anti-3-nitrotyrosine (anti-3-NTyr) immunoreactivity in Bsm and Pcm muscles. Appropriate statistical tests were done to determine significant differences among groups (P ≤ 0.05). RESULTS: The average CSA of Bsm was not significantly different, albeit a high percentage of myofibers was enlarged in late-pregnant and primiparous rabbits on P3; ß-glucuronidase activity and indirect parameter of muscle damage was also higher. These variables did not change in the Pcm muscle during the different reproductive stages. In contrast, the 3-NTyr immunoreactivity, an indicator of oxidative damage, was increased on P3 for Pcm myofibers and P20 for myofibers of both muscles. CONCLUSIONS: Our findings demonstrate reliable signs of damage to Bsm and Pcm muscles in young female rabbits passing different reproductive stages. Damage to the Bsm muscles as detected at the end of pregnancy persisted after delivery. This was not the case for Pcm muscles, in which damage seems to appear after delivery.

High Estradiol Differentially Affects the Expression of the Glucose Transporter Type 4 in Pelvic Floor Muscles of Rats.

PURPOSE: To characterize the relationship between serum estradiol levels and the expression of glucose transporter type 4 (Glut4) in the pubococcygeus and iliococcygeus muscles in female rats. METHODS: The muscles were excised from virgin rats during the metestrus and proestrus stages of the estrous cycle, and from sham and ovariectomized rats implanted with empty or estradiol benzoate-filled capsules. The expression of estrogen receptors (ERs) was inspected in the muscles at metestrus and proestrus. Relative Glut4 expression, glycogen content, and serum glucose levels were measured. Appropriate statistical tests were done to identify significant differences (P≤0.05). RESULTS: The pubococcygeus and iliococcygeus muscles expressed ERα and ERß. Glut4 expression and glycogen content in the pubococcygeus muscle were higher at proestrus than at metestrus. No significant changes were observed in the iliococcygeus muscle. In ovariectomized rats, the administration of estradiol benzoate increased Glut4 expression and glycogen content in the pubococcygeus muscle alone. CONCLUSION: High serum estradiol levels increased Glut4 expression and glycogen content in the pubococcygeus muscle, but not in the iliococcygeus muscle.

Multiparity affects conduction properties of pelvic floor nerves in rabbits.

INTRODUCTION: Women often develop pelvic floor dysfunction due to damage to the pelvic musculature during childbirth; however, the effect on pelvic floor nerves function is less understood. This study used adult rabbits to evaluate the electrophysiological and histological characteristics of the bulbospongiosus (Bsn) and pubococcygeus nerves (Pcn) in multiparity. METHODS: Compound nerve action potentials (CNAP) were compared between age-matched nulliparous and multiparous animals and associated to the histological characteristics of myelinated axons from the Bsn and Pcn nerves. The extensor digitorum longus nerve (EDLn) was used as negative control. Data were analyzed with unpaired two-tailed Student's t test or Mann-Whitney U test to determine significant differences between groups. RESULTS: The onset and peak latencies, duration, and conduction velocity of the motor fibers in these pelvic nerves were not significantly different between nulliparous and multiparous animals. However, the peak-to-peak amplitude and area of the CNAP in both Bsn and Pcn were reduced in multiparous rabbits. Histology showed a higher percentage of axons with myelin disorganization caused by multiparity in these pelvic nerves. Together, the data indicate a reduction in the number of functional pelvic axons due to multiparity. As expected, no effect of parity was observed in the EDLn controls. CONCLUSIONS: Present findings demonstrated that multiparity affects myelination and consequently conduction properties in the small pelvic floor nerves.

Hypothyroidism affects lipid and glycogen content and peroxisome proliferator-activated receptor δ expression in the ovary of the rabbit.

Dyslipidaemia and hyperglycaemia are associated with ovarian failure and both have been related to hypothyroidism. Hypothyroidism promotes anovulation and ovarian cysts in women and reduces the size of follicles and the expression of aromatase in the ovary of rabbits. Considering that ovarian steroidogenesis and ovulation depend on lipid metabolism and signalling, the aim of the present study was to analyse the effect of hypothyroidism on the lipid content and expression of peroxisome proliferator-activated receptor (PPAR) δ in the ovary. Ovaries from female rabbits belonging to the control (n=7) and hypothyroid (n=7) groups were processed to measure total cholesterol (TC), triacylglycerol (TAG) and glycogen content, as well as to determine the presence of granules containing oxidized lipids (oxysterols and lipofuscin) and the relative expression of perilipin A (PLIN-A) and PPARδ. Hypothyroidism increased TC and glycogen content, but reduced TAG content in the ovary. This was accompanied by a reduction in the expression of PLIN-A in total and cytosolic extracts, changes in the presence of granules containing oxidative lipids and low PPARδ expression. The results of the present study suggest that hypothyroidism modifies the content and signalling of lipids in the ovary, possibly affecting follicle maturation. These results could improve our understanding of the association between hypothyroidism and infertility in females.

Hypothyroidism impairs somatovisceral reflexes involved in micturition of female rabbits.

AIMS: To determine the impact of hypothyroidism on the bladder and urethral functions as well as in the activation of the pubococcygeous (Pcm) and bulbospongiosus (Bsm) during micturition. METHODS: Age-matched control and methimazole-induced hypothyroid female rabbits were used to simultaneously record cystometrograms, urethral pressure, and the reflex activation of Pcm and Bsm during the induced micturition. Urodynamic and urethral variables were measured. Activation or no activation of the Pcm and Bsm during the storage and voiding phases of micturition were categorized as 1 or 0. Significant differences (P ≤ 0.05) between control and hypothyroid groups were determined with unpaired Student-t or Mann-Whitney tests. RESULTS: One-month induced hypothyroidism increased the residual volume and threshold pressure while the opposite was true for the voided volume, maximal pressure, and voiding efficiency. Urethral pressure was also affected as supported by a notorious augmentation of the urethral resistance, among other changes in the rest of measured variables. Hypothyroidism also affected the reflex activation of the Pcm in the voiding phase of micturition. CONCLUSIONS: Our findings demonstrate hypothyroidism impairs the bladder and, urethral functions, and reflex activation of Pcm and Bsm affecting the micturition in female rabbits.