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.

Neural interrelationships of autonomic ganglia from the pelvic region of male rats.

The aims of the present study were to describe, in male rats, the anatomical organization of the major and accessory pelvic ganglia (MPG, AG; respectively), the interrelationship of the pelvic plexus components, and the morphometry of the pelvic postganglionic neurons. Anatomical, histochemical and histological studies were performed in anesthetized adult Wistar male rats. We found that the pelvic plexus consists of intricate neural circuits composed of two MPG, and three pairs of AG (AGI, AGII, AGIII) anatomically interrelated through ipsilateral and contralateral commissural nerves. Around 30 nerves emerge from each MPG and 17 from AGI and AGII. The MPG efferent nerves spread out preganglionic information to several pelvic organs controlling urinary, bowel, reproductive and sexual functions, while AG innervation is more regional, and it is confined to reproductive organs located in the rostral region of the urogenital tract. Both MPG and AG contain nerve fascicles, blood vessels, small intensely fluorescent cells, satellite cells and oval neuronal somata with one to three nucleoli. The soma area of AG neurons is larger than those of MPG neurons (p < 0.005). The MPG contains about 75% of the total pelvic postganglionic neurons. Our findings corroborated previous reports about MPG inputs, and add new information regarding pelvic ganglia efferent branches, AG neurons (number and morphometry), and neural interrelationship between the pelvic plexus components. This information will be useful in designing future studies about the role of pelvic innervation in the physiology and pathophysiology of pelvic functions.

Tissue alterations in urethral and vaginal walls related to multiparity in rabbits.

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.

Morphometry of paravaginal ganglia from the pelvic plexus: impact of multiparity, primiparity, and pregnancy.

OBJECTIVES: (1) To describe the morphology of paravaginal ganglia and the neurochemical pattern of their neurons in virgin rabbits. (2) To analyze the effects of multiparity, primiparity and late pregnancy on morphometry of ganglia and their neurons. STUDY DESIGN: The morphology and neurochemical pattern of paravaginal ganglia were described in virgin nulliparous Chinchilla breed rabbits. Acetylcholinesterase histochemistry, Masson's trichrome, and immunohistochemistry for cholinergic and adrenergic neurons, and estrogen receptors (ERs) were undertaken. The area covered by ganglia (ganglionic area), the number of neurons, and the neuron soma area were measured in multiparas (4 consecutive and successive deliveries) and age-matched nulliparas. The same variables were measured in primiparous, late-pregnant, and age-matched nulliparous rabbits. RESULTS: Paravaginal ganglia were adjoined to the dorsolateral walls of the pelvic vagina. Their neurons were mostly cholinergic and expressed ERα and ERß. Multiparity increased the ganglionic area and reduced the number of ganglionic neurons. Late pregnancy transiently reduced the neuron soma area, which was coincident with a low expression of ERs. CONCLUSION: Multiparity but not primiparity affected the morphometry of ganglia. The hormonal state present in late pregnancy alters the neuron soma area and ER expression. Our findings support the notion that reproduction influences the morphometry of the pelvic plexus.

General tissue characteristics of the lower urethral and vaginal walls in the domestic rabbit.

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.