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Estrogen receptor expression and neuronal nitric oxide synthase in the clitoris and prepucial gland structures of mice
ESPU Meeting 2007
S69
S13: Intersex 2 # S13-1 (PP)
ESTROGEN RECEPTOR EXPRESSION AND NEURONAL NITRIC OXIDE SYNTHASE IN THE CLITORIS AND PREPUCIAL GLAND STRUCTURES OF MICE Nieves MARTIN-ALGUACIL, Justine SCHOBER*, Lee-Ming KOWy and Donald PFAFFy The Rockefeller University, Dept of Neurobiology & Behavior, New York, USA, and Dept of Anatomy & Embriology, School of Veterinary Medicine, Univ. Complutense, Madrid, Spain, New York, USA - * Hamot Medical Center, Erie, USA, and Dept of Neurobiology & Behavior, The Rockefeller University, New York, USA, New York, USA - y The Rockefeller University, Department of Neurobiology and Behavior, New York, USA
PURPOSE ERa is the predominant form of estrogen receptor expressed in uterine and vaginal tissue of the murine female reproductive tract. Expression of both isoforms has been detected in vaginal tissue and in the ovary. Estrogen receptors were seen in basal and suprabasal cells of vaginal epithelium and epidermis of labia minora Understanding the locations of the different isoforms of the ER within genital structures may elucidate mechanisms of genital growth and development. It is well known that the sensory condition of the vulva epithelium is highly influenced by estrogen status with enlargement of the genitosensory field in response to this hormone. Estrogens may also prove crucial to the maintenance of urogential sensitivity. Their ability to promote the growth of neurons has been established. Abundant literature has reported on nitric oxide in the context of the internal female
reproductive tract and penis, few studies have investigated the female external genital structure or the clitoris. The aim of this work is to study the presence of estrogen receptors (ER), as well as n-NOS in the mouse clitoris.
MATERIALS AND METHODS Immunocytochemical studies specific for Estrogen Receptor-a, Estrogen Receptorb, and nNOS were performed on a series of sections of the pelvic area including the prepucial glands and clitoris of ten mice. Selected sections were also stained with Masson’s trichrome.
RESULTS ERa was expressed predominantly by the glandular epithelium as well as in the glandular tissue, preputial and apocrine
glands. ERa was observed in the nuclei of stromal cells around the cavernous tissue and near the epithelium of the clitoris. ERb immunostaining was only detected in few cells located in the vascular lumen of the cavernous tissue of the clitoris. nNOS immunostaining was detected in the clitoris, the preputial glans and the connective tissue.
CONCLUSIONS ERa, ERb isoforms and neuronal NOS are present in the clitoris and prepucial glands of female mice in different cellular localizations and with differing levels of receptivity. Functional studies would further elucidate the role of receptor functions and their relationship to neuronal expression of nitric oxide.
# S13-2 (PP)
THREE-DIMENSIONAL RECONSTRUCTION OF THE EPHRIN-B2 FEMALE MOUSE MODEL OF ANORECTAL MALFORMATIONS Juan PRIETO, Nilda GARCIA*, Maria MARTINEZy, Christopher DRAVISz, Dolores VASQUEZ{, Mark HENKEMEYERx and Linda BAKER University of Texas Southwestern Medical Center, Pediatric Urology, Dallas, USA - * University of Texas Southwestern Medical Center, Pediatric Surgery, Dallas, USA - y University of Texas Southwestern Medical Center, Research Pediatric Surgery and Urology, Dallas, USA - z University of Texas Southwestern Medical Center, Center for Development Biology and Kent Waldrep Center for Basic Research, Dallas, USA - { University of Texas Southwestern Medical Center, Urology, Dallas, USA - x University of Texas Southwestern Medical Center, Center for Developmental Biology and kent Waldrep Center for Basic Research, Dallas, USA
PURPOSE Persistent cloaca is a rare, severe congenital anomaly. The Eph family receptor tyrosine kinases and their ligands, the ephrin family, mediate diverse embryonic cell-cell recognition events. We previously reported ephrinB2lacZ/lacZ mice manifest male high imperforate anus and female persistent
cloaca (Dravis et al. Dev Biol 2004; 271(2): 272-290). We studied and demonstrated via 3D-reconstruction whether altered ephrin-B2 signaling in female ephrin-B2 lacZ/lacZ mutant mice affect apoptosis, thereby possibly contributing to persistent cloaca malformation.
MATERIAL AND METHODS Female embryonic days 11.5, 13.5 and 16.5 (E11.5, E13.5 and E16.5) wild type (WT), ephrin-B2lacZ/þ (Z/þ) and ephrinB2 lacZ/lacZ (Z/Z) mice sections were dual labeled for ephrin-B2 and TUNEL immunohistochemistry. Antigalactosidase antibody detected the ephrin-B2-gal fusion protein in Z/þ and
S69
S13: Intersex 2 # S13-1 (PP)
ESTROGEN RECEPTOR EXPRESSION AND NEURONAL NITRIC OXIDE SYNTHASE IN THE CLITORIS AND PREPUCIAL GLAND STRUCTURES OF MICE Nieves MARTIN-ALGUACIL, Justine SCHOBER*, Lee-Ming KOWy and Donald PFAFFy The Rockefeller University, Dept of Neurobiology & Behavior, New York, USA, and Dept of Anatomy & Embriology, School of Veterinary Medicine, Univ. Complutense, Madrid, Spain, New York, USA - * Hamot Medical Center, Erie, USA, and Dept of Neurobiology & Behavior, The Rockefeller University, New York, USA, New York, USA - y The Rockefeller University, Department of Neurobiology and Behavior, New York, USA
PURPOSE ERa is the predominant form of estrogen receptor expressed in uterine and vaginal tissue of the murine female reproductive tract. Expression of both isoforms has been detected in vaginal tissue and in the ovary. Estrogen receptors were seen in basal and suprabasal cells of vaginal epithelium and epidermis of labia minora Understanding the locations of the different isoforms of the ER within genital structures may elucidate mechanisms of genital growth and development. It is well known that the sensory condition of the vulva epithelium is highly influenced by estrogen status with enlargement of the genitosensory field in response to this hormone. Estrogens may also prove crucial to the maintenance of urogential sensitivity. Their ability to promote the growth of neurons has been established. Abundant literature has reported on nitric oxide in the context of the internal female
reproductive tract and penis, few studies have investigated the female external genital structure or the clitoris. The aim of this work is to study the presence of estrogen receptors (ER), as well as n-NOS in the mouse clitoris.
MATERIALS AND METHODS Immunocytochemical studies specific for Estrogen Receptor-a, Estrogen Receptorb, and nNOS were performed on a series of sections of the pelvic area including the prepucial glands and clitoris of ten mice. Selected sections were also stained with Masson’s trichrome.
RESULTS ERa was expressed predominantly by the glandular epithelium as well as in the glandular tissue, preputial and apocrine
glands. ERa was observed in the nuclei of stromal cells around the cavernous tissue and near the epithelium of the clitoris. ERb immunostaining was only detected in few cells located in the vascular lumen of the cavernous tissue of the clitoris. nNOS immunostaining was detected in the clitoris, the preputial glans and the connective tissue.
CONCLUSIONS ERa, ERb isoforms and neuronal NOS are present in the clitoris and prepucial glands of female mice in different cellular localizations and with differing levels of receptivity. Functional studies would further elucidate the role of receptor functions and their relationship to neuronal expression of nitric oxide.
# S13-2 (PP)
THREE-DIMENSIONAL RECONSTRUCTION OF THE EPHRIN-B2 FEMALE MOUSE MODEL OF ANORECTAL MALFORMATIONS Juan PRIETO, Nilda GARCIA*, Maria MARTINEZy, Christopher DRAVISz, Dolores VASQUEZ{, Mark HENKEMEYERx and Linda BAKER University of Texas Southwestern Medical Center, Pediatric Urology, Dallas, USA - * University of Texas Southwestern Medical Center, Pediatric Surgery, Dallas, USA - y University of Texas Southwestern Medical Center, Research Pediatric Surgery and Urology, Dallas, USA - z University of Texas Southwestern Medical Center, Center for Development Biology and Kent Waldrep Center for Basic Research, Dallas, USA - { University of Texas Southwestern Medical Center, Urology, Dallas, USA - x University of Texas Southwestern Medical Center, Center for Developmental Biology and kent Waldrep Center for Basic Research, Dallas, USA
PURPOSE Persistent cloaca is a rare, severe congenital anomaly. The Eph family receptor tyrosine kinases and their ligands, the ephrin family, mediate diverse embryonic cell-cell recognition events. We previously reported ephrinB2lacZ/lacZ mice manifest male high imperforate anus and female persistent
cloaca (Dravis et al. Dev Biol 2004; 271(2): 272-290). We studied and demonstrated via 3D-reconstruction whether altered ephrin-B2 signaling in female ephrin-B2 lacZ/lacZ mutant mice affect apoptosis, thereby possibly contributing to persistent cloaca malformation.
MATERIAL AND METHODS Female embryonic days 11.5, 13.5 and 16.5 (E11.5, E13.5 and E16.5) wild type (WT), ephrin-B2lacZ/þ (Z/þ) and ephrinB2 lacZ/lacZ (Z/Z) mice sections were dual labeled for ephrin-B2 and TUNEL immunohistochemistry. Antigalactosidase antibody detected the ephrin-B2-gal fusion protein in Z/þ and