Anatomic Variations of the Perineal Arteries and Nerves in the Male and Female Dog and Its Clinical Implications.

The anatomic variations of the perineal arteries and nerves are studied in the dog. The aim of the study is to provide a more detailed understanding of the blood supply and innervation of the perineal region, providing detailed information on the perineal arteries and nerve distribution and their variability in male and female dogs. The study used 232 pelvic halves from 116 adult dogs and analysed the differences using the chi-squared test. The results showed that the presumptive model for perineal artery distribution described in N.A.V. was observed in 46% of the specimens. Additionally, a dorsal perineal artery "long type" was found in 13% of the dogs, and a perineal trunk was present in 41% of the dogs. In the study, there was no variation in perineal nerve distribution, and it was found that the perineal nerve did not run together with the dorsal perineal artery when it was the "short type" as described in the presumptive model for perineal artery distribution. Instead, it always followed the route of the dorsal perineal artery "long type" and the perineal trunk when they were present. The findings of the study may be useful for veterinary surgeons when approaching the perineal region in dogs.

Anatomy-based diagnostic criteria for complex body wall anomalies (CBWA).

BACKGROUND: Precise diagnosis and classification of CBWA cases can be challenging. BSA are considered when there is a body wall anomaly, skeletal abnormalities, and the umbilical cord is anomalous, absent or rudimentary, and LBWC when there is a body wall and structural limb anomalies with or without craniofacial abnormalities. METHODS: PubMed was searched for body stalk anomalies, limb body wall complex, body stalk anomalies and amniotic band syndrome, and limb body wall complex and amniotic band syndrome. Sixty nine articles were selected and reviewed. This article systematically classifies the variants of CBWA in 218 cases, the study is based on the embryological and anatomical criteria established by Martín-Alguacil and Avedillo to study BSA in the pig. RESULTS: Eight different BSA presentation were defined. One hundred and eighty nine cases were classified as BSA, from which five were Type I, nine Type II, 20 Type III, 57 Type IV, 11Type V, 24 Type VI, 11 Type VII, and 52 Type VIII. Twenty six cases presented cranial phenotype, 114 abdominal phenotype, 42 cranio/abdominal overlapping phenotype, and five without defined phenotype. In addition, 52 BSA cases presented some kind of spinal dysraphism (SPDYS) and were classified as BSA/SPDYS, most of these cases did not show structural limb anomalies, except for three cases and were classified as LBWC/SPDYS. CONCLUSION: This morphology-based classification represents a useful tool for clinical diagnosis, it helps to quantify and to evaluate CBWA in a precise, objective manner.

Body stalk anomalies in pig-Definition and classification.

The presence of body wall closing defects (abdominoschisis and thoracoabdominoschisis) in combination with other congenital malformations was studied in the pig (Sus scrofa domesticus). After clinical examination and literature review, body wall defects with multiple congenital anomalies in eight pigs were described, and classified using anatomical and embryological criteria. Several BSA presentations were identified and classified as follows: (a) BSA Type I: fetus with spinal and UC defects, thoracoabdominoschisis, anal atresia and/or other internal organs structural defects, and structural limb defects; (b) BSA Type II: fetus with spinal and UC defects, thoracoabdominoschisis, anal atresia and/or other internal organs structural defects, and nonstructural limb defects; (c) BSA Type III: fetus with spinal and UC defects, abdominoschisis, anal atresia and/or other internal organs structural defects, and structural limb defects; and (d) BSA Type IV: fetus with spinal and UC defects, abdominoschisis, anal atresia and/or other internal organs structural defects, and nonstructural limb defects. Two types of LBWC were differentiated: LBWC Type I: characterized by thoracoabdominoschisis and structural limb defects, and LBWC Type II: characterized by abdominoschisis and structural limb defects, corresponding to BSA type I and type III. This is the first report on BSA and LBWC in the pig.

Body wall defects and amniotic band syndrome in pig (Sus scrofa domesticus).

The amniotic band syndrome is a congenital condition. It is characterized by the presence of fibrous amniotic bands that may entangle or entrap different foetal parts in utero, resulting in deformation, malformation or disruption. We report on a female piglet presenting amniotic band adherences in the right abdominal flank, several body wall defects (gastroschisis, abdominoschisis with omphalocele), severe scoliosis, anomalous umbilical cord with single umbilical artery, anal atresia, anomalous liver and absent gall bladder, hypoplastic genitalia, ankylosis and arthrogryposis in pelvic limbs, and bilateral patellar agenesia. The ethiopatogenia is discussed, as well as the comparative and embryological implications.

VACTERL Association in a Female Pig (Sus scrofa domesticus).

VACTERL/VATER association is a condition defined by the presence of at least three of the following congenital malformations: vertebral defects (V), anal atresia (A), cardiac defects (C), tracheo-oesophageal fistula (TE), renal anomalies (R) and limb abnormalities (L). We describe a stillborn female piglet with cardiac anomalies, renal defects, vertebral anomalies, anal atresia and a single umbilical artery (SUA), which are the main features of VACTERL association. In addition, the piglet had a unilateral abdominal wall defect. This was the only affected animal in a litter of 16 piglets. The molecular inductive mechanisms of this disorder are discussed, as well as the comparative and embryological implications.

Analyses of rapid estrogen actions on rat ventromedial hypothalamic neurons.

Rapid estrogen actions are widely diverse across many cell types. We conducted a series of electrophysiological studies on single rat hypothalamic neurons and found that estradiol (E2) could rapidly and independently potentiate neuronal excitation/depolarizations induced by histamine (HA) and N-Methyl-d-Aspartate (NMDA). Now, the present whole-cell patch study was designed to determine whether E2 potentiates HA and NMDA depolarizations - mediated by distinctly different types of receptors - by the same or by different mechanisms. For this, the actions of HA, NMDA, as well as E2, were investigated first using various ion channel blockers and then by analyzing and comparing their channel activating characteristics. Results indicate that: first, both HA and NMDA depolarize neurons by inhibiting K(+) currents. Second, E2 potentiates both HA and NMDA depolarizations by enhancing the inhibition of K(+) currents, an inhibition caused by the two transmitters. Third, E2 employs the very same mechanism, the enhancement of K(+) current inhibition, thus to rapidly potentiate HA and NMDA depolarizations. These data are of behavioral importance, since the rapid E2 potentiation of depolarization synergizes with nuclear genomic actions of E2 to facilitate lordosis behavior, the primary female-typical reproductive behavior.

Terminal innervation of the male genitalia, cutaneous sensory receptors of the male foreskin.

PURPOSE: Understanding the types of sensory nerve termini within the glabrous skin of the human male foreskin could throw light on surgical outcomes and therapeutic possibilities for the future. Various receptor types sense changes in temperature, position, pressure, pain, light touch, itch, burning and pleasurable sexual sensations. Similarities and differences in innervation characteristics and density might become apparent when the glans penis is compared with homologous structures in the female genitalia. The aim of this study is to document the presence and characteristics of cutaneous sensory receptors in the human penile foreskin using a histopathological study of the nerve termini to achieve a more complete understanding of sensory experiences. METHODS: Foreskin samples were obtained from ten boys (aged 1-9 years) who had undergone circumcision. Informed consent was obtained from the parent/legal guardian. The samples were examined after modified Bielschowsky silver impregnation of neural tissue, and immunocytochemistry against gene protein product (PGP) 9.5 and neuron-specific enolase (NSE). RESULTS: PGP 9.5 appeared to be the most sensitive neural marker. Free nerve endings were identified in the papillary dermis visualized as thin fibers, mostly varicose, with either branched or single processes, either straight or bent. Two types of sensory corpuscle were identified: capsulated and non-capsulated. Meissner-like corpuscles were located in the papillary dermis. Capsulated corpuscles resembled typical Pacinian corpuscles, comprising a single central axon surrounded by non-neural periaxonic cells and lamellae. The capsulated corpuscles were strongly positive for PGP 9.5 and NSE. CONCLUSIONS: Free nerve endings, Meissner's corpuscles and Pacinian corpuscles are present in the human male foreskin and exhibit characteristic staining patterns.

Terminal innervation of female genitalia, cutaneous sensory receptors of the epithelium of the labia minora.

INTRODUCTION: Little information is available regarding the sensory nerve endings within the glabrous skin of the external female genitalia. The diversity of possible sensations suggests a variety of receptor types. Comprehensive knowledge of the sensory stimuli, including stimulus position, changes in temperature, pressure and pain, is critical for addressing pain and sexual function disorders clinically. The aim of this neuro-histological study is document the presence and characteristics of cutaneous sensory receptors in female genital tissue. MATERIALS AND METHODS: Labial skin samples were obtained from ten normal girls (aged 1-9 years). The specimens were waste tissue obtained during surgical intervention. They were all obtained by the senior investigator, a pediatric urologist, after the parent or legal guardian had given informed consent. The specimens were stained by Cajal-type silver impregnation and by immunocytochemistry against protein gene product (PGP) 9.5 and neuron-specific enolase (NSE). RESULTS: PGP 9.5 was the most sensitive neural marker for identifying cutaneous sensory receptors. Free nerve endings (FNEs) in the papillary dermis appeared as thin fibers, varicose, branched or single processed, straight or bent. In the labia minora, FNEs were identified in the strata basale, spinosum and granulosum of the epidermis. Non-capsulated (Meissner-like) corpuscles in the dermal papillae interdigitated with epidermal ridges of the skin. Capsulated corpuscles protruded from the deep dermis into the epidermis. Encapsulated corpuscles and cells located in the inner and outer cores were strongly positive for PGP 9.5. CONCLUSIONS: FNEs, Meissner's corpuscles and Pacinian corpuscles are present in the female labia minora and exhibit characteristic staining patterns.

Immunocytochemical characterization of pacinian-like corpuscles in the labia minora of prepubertal girls.

STUDY OBJECTIVE: To better understand the precise role of sensory corpuscles within the female external genitalia. DESIGN: After IRB approval, waste tissue samples were obtained from 10 normal girls (aged 2-9 years) who underwent surgery for labial fusion. Immunocytochemistry against protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), vasoactive intestinal peptide (VIP), 5-hydroxytryptamine transporter (5HTT), 5-hydroxytryptamine receptor 1A (5HT1A), Neuronal Peptide Y (NPY), neuronal nitric oxide synthase (nNOS), and estrogen receptors (ER) α and ß was performed. RESULTS: Pacinian-like corpuscles were identified in epithelium of labia minora of prepubertal girls. A central structure composed of an axon surrounded by a central core, outer core, external capsule, surrounded by encapsulated stroma, and a subsidiary innervation in the outer aspect of the corpuscle stroma stained for PGP 9.5 in the outer core and layers of the external capsule, NSE positive cells in layers of the outer core, 5HTT in stroma of the corpuscle and cells located in layers of the outer core, 5HT1A in cells of outer core, NPY in stroma of the corpuscle, and nNOS in external core and external capsule of the central structure. ERα was present in stroma, external core, and external capsule, and ERß in stroma of the corpuscle with subsidiary innervation in the stroma positive to PGP 9.5, VIP, and NPY. CONCLUSION: PGP 9.5, NSE, ERα, nNOS, and 5HTT immunoreaction detected in the outer core and external capsule could indicate these areas may play an important role in the functional aspects of the Pacinian-like corpuscle.

Innervation of the labia minora of prepubertal girls.

INTRODUCTION: Surgical and histologic sources of information give little reference to innervation, vascular, and epithelial details of the labia minora. Little is known about areas of nerve density, epithelial qualities, and vascular compartments of the labial minora that contribute to sexual arousal and orgasm. Surgical procedure development and counsel about surgical risks related to labioplasty and surgical flaps created from labial tissue may be based on inadequate information. METHODS: Labial samples from 10 normal girls (aged 2-9 years) who underwent surgery for labial fusion utilized waste tissue strips for immunohistochemical identification of S-100 and neuronal nitric oxide synthase (nNOS) in the labia minora. RESULTS: Vascular and lymphatic plexus lie within the reticular dermis, which contains a dense mesh of nerve fibers with a higher concentration of nerve fiber at the level of the subepithelial plexus. Dense innervations are located at the epidermis, extending along the basal and spinous layers of the epithelium of labia minora. Nerve bundles in the papillary dermis are associated with sebaceous and eccrine glands and nerve terminals located throughout the epithelium. The introital epithelium of the labia minora is highly innervated with widespread and intense staining, detected in the introital border of the labia minora versus the external one. The dermis appeared to display S-100 and nNOS immunolabelling. S-100 was also immunopositive in the epidermis. CONCLUSION: Labia minora is highly innervated along its entire edge. Related vascular compartment tissue involved in engorgement during sexual arousal makes this tissue important for sexual response. Labioplasty risks removal of tissue with an important contribution to sensory sexual arousal. Movement of labial tissue during genitoplasty may have different sensory outcomes dependent on which labial surface is used.

Success of treatment modalities for labial fusion: a retrospective evaluation of topical and surgical treatments.

STUDY OBJECTIVE: Standard treatment for girls with labial fusion has included topical estrogen cream, manual separation, or surgery. Side effects may limit the use of topical estrogen. Betamethasone has recently shown efficacy at separating labial fusion. Local irritation and inflammation may be an initiator of labial fusion. No adverse effects of betamethasone treatment have been documented. Long-term side effects are unknown. This study compares therapies for conservative management of labial fusion for efficacy and focuses on the response rate, time to separation, recurrence, and side effects of treatment. DESIGN: A retrospective chart review. PARTICIPANTS: One hundred fifty-one prepubertal girls, mean age 3 years (range 0.25-8.75 years) diagnosed with labial fusion. MAIN OUTCOME MEASURES: To investigate the incidence of related symptoms, length of topical estrogen or betamethasone treatment, side effects, rate of successful separation, rates of recurrence, percentage requiring surgery, and postoperative outcomes in patients with labial adhesion who underwent treatment. RESULTS: Of 151 patients with labial adhesion, 11 (7.3%) presented with urinary frequency, 30 (19.9%) with urinary tract infections, 13 (8.6%) with vaginitis, and 19 (12.6%) with post-void dripping. When compared to patients treated with betamethasone (1.3 months), patients treated primarily with premarin took nearly twice as long (2.2 months) for resolution of their adhesions. Rates of recurrence were lower for patients receiving betamethasone therapy. Side effects for estrogen therapy included breast budding and vaginal bleeding, and for betamethasone, local irritation was reported. Some patients went on to surgery and experienced recurrence after surgery. CONCLUSION: Initial comparison of topical estrogen and betamethasone treatment of labial fusion suggests that betamethasone may separate fusion quicker with less recurrence and fewer side effects than topical estrogen therapy.

Oestrogen receptor expression and neuronal nitric oxide synthase in the clitoris and preputial gland structures of mice.

OBJECTIVE: To study the presence of oestrogen receptors (ER) and neuronal nitric oxide synthase (nNOS) in the mouse clitoris. MATERIALS AND METHODS: A series of sections of the pelvic area, including the preputial glands and clitoris, of 10 mice were assessed by immunocytochemical studies specific for ER-alpha and -beta, and nNOS; selected sections were also stained with Masson's trichrome. RESULTS: ER alpha was detected in the epithelium of the gland of the clitoris, and in the glandular tissue, preputial and apocrine gland. ER alpha was detected in the nuclei of stromal cells around the cavernous tissue and near the epithelium of the clitoris. Cytoplasm ER alpha was detected in a few cells in an area ventral to the clitoral gland. There was also nuclear staining in the connective tissue cells surrounding the clitoris. Very light ER beta immunostaining was detected in the clitoris and in the tissue related to it. There were some cells with nuclear staining in the vessels of the cavernous tissue of the clitoris. nNOS immunostaining was detected in the clitoris, the preputial gland and the connective tissue. CONCLUSION: ER alpha and beta isoforms, and nNOS, are present in the clitoris and preputial glands of female mice in different cellular locations and with differing levels of receptivity. Functional studies would further elucidate the role of receptor functions and their relationship to the neuronal expression of NO.

Clitoral sexual arousal: neuronal tracing study from the clitoris through the spinal tracts.

PURPOSE: Although genital tactile stimulation is regarded as a precursor to sexual arousal and a recognized initiator of central nervous system arousal, specific afferent neural pathways transmit sensory stimuli of arousal, beginning at the epithelial level on the clitoris and following the course of arousal stimuli through the central nervous system. Limited knowledge exists of the pathway from the cutaneous receptors of nerves originating in the epithelial tissue of the clitoris and continuing to spinal cord afferents. Such information may contribute to an understanding of sexual arousal, particularly in female vertebrates. We further defined the neural pathways and mechanisms responsible for arousal originating in the epithelium of the clitoris as well as related neural pathways to the spinal cord in a murine model. MATERIALS AND METHODS: We performed a comprehensive review of the published relevant clinical and histological material from human and nonhuman vertebrate studies. In 29 adult female C57B1/6 mice the distribution of pelvic nerves and vessels was mapped. Gross dissection of 4 female mice was facilitated by resin injection of the vascular system in 2. Neuronal tracing was performed in 25 mice that received clitoral injection of wheat germ agglutinin-horseradish peroxidase into the clitoris and were sacrificed after 72 to 96 hours. The spinal cord and periclitoral tissue were removed and fixed. Immunohistochemistry was performed. RESULTS: Gross anatomy of the mouse clitoris showed that pudendal and hypogastric nerves have a major role in the innervation of the external genitalia. Neuronal tracing revealed that the greatest nerve density was noted in the L5/6 spinal cord. The distribution extended from S1 to L2 with no labeling seen in the L3 spinal cord. Wheat germ agglutinin-horseradish peroxidase labeling was seen caudal in levels S1 through L4 and rostral in L2. CONCLUSIONS: Understanding the neuroanatomy of the clitoris using a murine model may provide a valuable tool for the study of sexual arousal disorders and the further understanding of sexual function related to neural pathologies and trauma.

Oestrogen receptors and their relation to neural receptive tissue of the labia minora.

OBJECTIVE: To assess the cellular distributions of oestrogen receptors alpha and beta (ER alpha and ER beta) and neuronal nitric oxide synthase (nNOS) in the labia minora, as knowledge about ER type and function may clarify the role of oestrogens in vaginal scar formation and improve outcomes in female genital surgery. SUBJECTS AND METHODS: Labial samples were taken from 10 girls (aged 2-9 years) who underwent surgery for labial fusion. The waste tissue strips obtained were used for immunohistochemical identification of ER alpha and ER beta, and nNOS in the labia minora. RESULTS: There was ER alpha nuclear staining in the stroma of the labia minora close to the clitoris, and basal and suprabasal in the epidermal cells membrane restricted to superficial sections of the labia minora. ER beta was found in the stroma of the labia minora closer to the clitoris and in superficial sections, in the basal epidermal cells membrane and apocrine glandular epithelial cells membrane. There was also ER beta cell membrane staining in the basal and suprabasal epithelial cells and fibroblasts in the lamina propria. CONCLUSIONS: Established ER presence allows the consideration of the introitus of the vagina as a target for oestrogen therapy in various clinical and surgical situations. Continuing elucidation of the immunohistochemistry of this external genital tissue might assist in the development of molecular tools to treat genital abnormalities. Details of this immunohistochemistry may also advance the understanding of the effects of sexual differentiation on the brain and other organ systems.

Clitoral sexual arousal: an immunocytochemical and innervation study of the clitoris.

OBJECTIVE: To further define neural pathways and mechanisms responsible for the arousing properties of the epithelium of the clitoris as well as related neural pathways associated with sexual arousal in a murine model. MATERIALS AND METHODS: Adult female C57B1/6 mice were used for gross dissection, facilitated by resin injection of the vascular system, and silver staining, and immunostaining for S-100 and neuronal nitric oxide synthase (nNOS). We also comprehensively reviewed relevant published clinical and histological material from both human and non-human vertebrate studies. RESULTS: The distal innervation consists of three nerve bundles: one related to the perineal region, one through the corpus cavernosum, and the third between the dorsal part of the clitoris and the urethra. Communicating nerve fibres were identified between the perineal, the corpus cavernous nerve (CN) and the dorsal nerve of the clitoris. Immunostaining for nNOS showed that the CN sends nNOS-positive fibres to join the dorsal nerve of the clitoris. In the same distal area of the clitoris, the connecting branches between the perineal nerve and the dorsal nerve of the clitoris are also nNOS positive. CONCLUSIONS: A rich network of nerve bundles and terminal branches were identified and associated with nNOS immunostaining in the cavernosal tissue of the body of clitoris. NO control of vasodilatation and neuronal signalling between the CN and the dorsal nerve of the clitoris could contribute to the engorgement and subsidence of clitoral tissue. This supports the initiation of sexual arousal by tactile stimuli. The distribution pattern of the general and peptidergic innervation in the murine clitoris is similar to that of the penis.

Significance of topical estrogens to labial fusion and vaginal introital integrity.

PURPOSE: One theory for labial fusion is low prepubertal estrogen levels. Topical estrogens remains the mainstay of therapy. Some patients require surgical lysis of the adhesion. Estrogen's action in regard to collagen may influence recurrent adhesions and adhesions that form after manual disruption or surgical separation. This study assesses the efficacy of topical estrogen to separate the labia, recurrence, and estrogen related side effects. Estrogens may have a role in vaginal healing in genital surgery. MATERIALS AND METHODS: Retrospective chart review of 109 girls from 3 months to 10 years old (mean 44 months) who had labial fusion. Data was collected on the length of topical estrogen treatment, rate of successful separation, rate of recurrence, percentage requiring surgery, and post-operative outcomes. RESULTS: Mean length of topical estrogen treatment was 3.7 months (range 0 to 36 months), with separation in 79% (85/107) of patients. Forty-one percent (44/107) had recurrence of labial fusion one to five times (range 2 to 72 months). Surgery was required in 21% (22/107). Ten percent of patients (2/21) had recurrence of labial fusion post-operatively (age 6,10 years, at 1, 18 month post-operatively). Neither of these two patients was responsive to topical estrogen cream post-operatively and required a second surgical separation. Side effects of estrogen were minimal breast development in 6 children and vaginal bleeding in one child. Discontinuance resulted in resolution of side effects. CONCLUSION: Topical estrogens were effective treatment for labial fusion. After surgical separation of adhesions there was a 10% recurrence rate. Estrogen therapy initiated after surgical failure was unsuccessful.

Arousing properties of the vulvar epithelium.

PURPOSE: The initiation of genital tactile stimulation is regarded as a precursor to sexual arousal and perhaps in women it is the most easily recognized initiator of central nervous system arousal. Unfortunately little published material details the specific mechanisms preceding arousal, beginning at the epithelial level, which are the sensory precursors to arousal. Little is known about its cutaneous receptors, nerves and the other histochemical properties of this epithelial tissue that contribute to sexual arousal. Sexual sensitivity evaluations target female genital somatosensory pathways for cutaneous sensation by testing evoked potentials of nerves, hot/cold and vibratory sensory discrimination. The anatomical bases of these several sensibilities form a subject for future investigation. We reviewed the known influences and mechanisms responsible for the arousing properties of the epithelium in the female external genitalia as well neural pathways associated with sexual arousal originating from the vulvar epithelium. MATERIALS AND METHODS: A comprehensive review was done of published, relevant clinical and histological material in human and nonhuman vertebrate studies. RESULTS: Tactile stimulation of the vulvar epithelium initiates changes suggesting complex integrative mechanisms. Influences of skin temperature, hormonal environment, mechanical tissue compliance and inflammation as well as the large number of transmitters and neuropeptides involved in peripheral pathways serving female sexual arousal speak of a direct sensory role. CONCLUSIONS: Genital epithelial cells may actively participate in sensory function to initiate sexual arousal by expressing receptors and releasing neurotransmitters in response to stimuli, resulting in epithelial-neuronal interactions.