Morphology and histology of the male reproductive tract of Caecilia thompsoni (amphibia: Gymnophiona).

We studied the male reproductive tract of individuals of different body sizes of Caecilia thompsoni to describe morphological characteristics in comparison to other Gymnophiona. The reproductive tract consists of paired testes segmented into chains of primary and secondary lobes, sperm ducts that empty to Wolffian ducts, the cloaca that receives the Wolffian ducts and possesses a phallodeum. Müllerian ducts are present and develop into paired glands that empty into the cloacal urodeum. Testicular secondary lobes contain lobules with cysts of the entire germinal cell line, whereas primary lobes, in the terminal ends of the chains, only have spermagonia, Sertoli cells, and connective tissue. The smallest individual examined (21 cm body length) was immature and only possessed a few testicular primary lobes. Once the individuals reach sexual maturity, the morphological characteristics are quite consistent at macroscopic and histological level among males of very different body sizes. The histological features of the Wolffian and Müllerian glands suggest a complementary secretory role between the two ducts. In the cloaca we found the propulsor muscle, venous sinuses, and blind sacs in the phallodeum, which differentiate C. thompsoni from other species of the genus. Despite these slight differences, the general morphological characteristics, both macroscopic and microscopic, of the reproductive tracts of adult males of C. thompsoni follow the pattern known for the reproductively active males of Gymnophiona.

Testosterone-induced "Virilization" of Mesonephric Duct Remnants and Cervical Squamous Epithelium in Female-to-Male Transgenders: A Report of 3 Cases.

Mesonephric ducts regress in genotypic females, leaving behind few remnants. These vestigial structures are often recognized in the mesosalpinx and paracervical regions. We report here 3 cases of female-to-male transgenders who underwent hysterectomy following testosterone treatment. Both female and male genital structures were identified on histologic examination. Although the morphologic appearances of the specimens were unremarkable, histologically 1 case revealed a well-formed fallopian tube as well as an epididymis and 2 cases showed prostate glands to be present in the cervical squamous epithelium.

Quiste del conducto de Gartner asociado a carcinoma microinvasor de cérvix y presentado en una paciente como prolapso de órganos pélvicos / Gartner's duct cyst associated with invasive cervical carcinoma mimicking pelvic organ prolapse

La incidencia de los quistes vaginales está estimada en un 1%; de estos, una parte los constituyen los quistes de origen embrionario, como es el caso del quiste del conducto de Gartner. Estos últimos suelen ser de pequeño tamaño y asintomáticos, aunque en algunas situaciones aumentan de tamaño y, por la localización, pueden dar lugar a sintomatología relacionada con patología del suelo pélvico. La importancia radica en un correcto diagnóstico diferencial, donde las pruebas complementarias de elección serán la ecografía transvaginal y, sobre todo, la resonancia magnética, la cual permitirá establecer las características del quiste, la localización y si existe o no asociación a anormalidades del sistema urinario. En este trabajo presentamos un caso de quiste del conducto de Gartner asociado a carcinoma invasivo de cérvix, diagnosticado en una mujer de 49 años que acudió a nuestra consulta presentando sintomatología relacionada con prolapso de órganos pélvicos, concretamente cistocele (AU)

The incidence of vaginal cysts is estimated to be 1%. Some of these cysts, such as Gartner's duct cyst, are of embryological origin. These cysts are usually small and asymptomatic but they can sometimes grow and, due to their location, produce symptoms related to pelvic floor disease. A correct differential diagnosis is essential, in which the main complementary tests are transvaginal ultrasound and especially magnetic resonance imaging, which will establish the features and location of the cyst as well as any anomalies of the urinary system. We report a case of Gartner's duct cyst associated with invasive cervical carcinoma in a 49-year-old woman who presented with symptoms similar to pelvic organ prolapse, specifically cystocele (AU)

The testicular sperm ducts and genital kidney of male Ambystoma maculatum (Amphibia, Urodela, Ambystomatidae).

The ducts associated with sperm transport from the testicular lobules to the Wolffian ducts in Ambystoma maculatum were examined with transmission electron microscopy. Based on the ultrastructure and historical precedence, new terminology for this network of ducts is proposed that better represents primary hypotheses of homology. Furthermore, the terminology proposed better characterizes the distinct regions of the sperm transport ducts in salamanders based on anatomy and should, therefore, lead to more accurate comparisons in the future. While developing the above ontology, we also tested the hypothesis that nephrons from the genital kidney are modified from those of the pelvic kidney due to the fact that the former nephrons function in sperm transport. Our ultrastructural analysis of the genital kidney supports this hypothesis, as the basal plasma membrane of distinct functional regions of the nephron (proximal convoluted tubule, distal convoluted tubule, and collecting tubule) appear less folded (indicating decreased surface area and reduced reabsorption efficiency) and the proximal convoluted tubule possesses ciliated epithelial cells along its entire length. Furthermore, visible luminal filtrate is absent from the nephrons of the genital kidney throughout their entire length. Thus, it appears that the nephrons of the genital kidney have reduced reabsorptive capacity and ciliated cells of the proximal convoluted tubule may increase the movement of immature sperm through the sperm transport ducts or aid in the mixing of seminal fluids within the ducts.

Molecular characteristics and alterations during early development of the human vagina.

Unresolved questions remain concerning the derivation of the vagina with respect to the relative contributions from the Müllerian ducts, the urogenital sinus, and the Wolffian ducts. Recent molecular and cellular studies in rodents have opened up a large gap between the level of understanding of vaginal development in mice and understanding of human vaginal development, which is based on histology. To compare the findings in mice with human vaginal development and to address this gap, we analysed molecular characteristics of the urogenital sinus, Wolffian ducts, and Müllerian ducts in 8-14-week-old human specimens using immunohistochemical methods. The monoclonal antibodies used were directed against cytokeratin (CK) 14, CK19, vimentin, laminin, p63, E-cadherin, caspase-3, Ki67, HOX A13, and BMP-4. The immunohistochemical analysis revealed that, during weeks 8-9, the epithelium of the Müllerian ducts became positive for p63 as p63-positive cells that originated from the sinus epithelium reached the caudal tip of the fused Müllerian ducts via the Wolffian ducts. The lumen of the fused Müllerian ducts was closed by an epithelial plug that contained both vimentin-positive and vimentin-negative cells. Subsequently, the resulting epithelial tube enlarged by proliferation of basal p63-positive cells. The first signs of squamous differentiation were detected during week 14, with the appearance of CK14-positive cells. According to our results, all three components, namely, the urogenital sinus, Wolffian ducts, and Müllerian ducts, interacted during the formation of the human vagina. The sinus epithelium provided p63-positive cells, the Wollfian ducts acted as a 'transporter', and the Müllerian ducts contributed the guiding structure for the vaginal anlagen. Epithelial differentiation began at the end of the period studied and extended in a caudo-cranial direction. The present study is one of the first to provide up-to-date molecular correlates for human vaginal development that can be compared with the results of cell biological studies in rodents.

PAX8 and PAX2 immunostaining facilitates the diagnosis of primary epithelial neoplasms of the male genital tract.

PAX8 and PAX2 are cell-lineage-specific transcription factors that are essential for the development of Wolffian and Müllerian ducts and have recently emerged as specific diagnostic markers for tumors of renal or Müllerian origin. Little is known about their expression in the Wolffian duct-derived human male genital tract. We report our findings of PAX8 and PAX2 expression in the epithelium of the normal male genital tract and in epithelial tumors derived therefrom using immunohistochemistry (IHC). We found that PAX8 and PAX2 were expressed in the epithelium of the male genital tract from the rete testis to the ejaculatory duct. Rare glands in the prostatic central zone, a tissue of purported Wolffian duct origin, were focally positive for PAX2, but no PAX8 was detected in this area, a finding that may warrant further study. We found diffuse expression of PAX8 and PAX2 in 1 case each of serous cystadenoma of the epididymis, carcinoma of the rete testis, Wolffian adnexal tumor of the seminal vesicle, and endometrioid carcinoma of the seminal vesicle. Neither PAX8 nor PAX2 was detected in the seminiferous tubules and interstitium of the normal testis, nor in Leydig cell tumors (n=6), Sertoli cell tumors (n=2), or 48 of 49 germ cell tumors. One pediatric yolk sac tumor showed focal and weak staining for PAX8. Tumors of mesothelial origin, that is, adenomatoid tumors (n=3) and peritoneal malignant mesotheliomas (n=37) in men, were negative for PAX2 and PAX8. Neither PAX2 nor PAX8 was present in other areas of the prostate. Expression of PAX8 and PAX2 in these primary epithelial neoplasms of the male genital tract is due to their histogenetic relationship with Wolffian or Müllerian ducts. PAX8 and PAX2 IHC may facilitate the diagnosis of these tumors and should be included in the differential diagnostic IHC panel.

The pelvic kidney of male Ambystoma maculatum (Amphibia, urodela, ambystomatidae) with special reference to the sexual collecting ducts.

This study details the gross and microscopic anatomy of the pelvic kidney in male Ambystoma maculatum. The nephron of male Ambystoma maculatum is divided into six distinct regions leading sequentially away from a renal corpuscle: (1) neck segment, which communicates with the coelomic cavity via a ventrally positioned pleuroperitoneal funnel, (2) proximal tubule, (3) intermediate segment, (4) distal tubule, (5) collecting tubule, and (6) collecting duct. The proximal tubule is divided into a vacuolated proximal region and a distal lysosomic region. The basal plasma membrane is modified into intertwining microvillus lamellae. The epithelium of the distal tubule varies little along its length and is demarcated by columns of mitochondria with their long axes oriented perpendicular to the basal lamina. The distal tubule possesses highly interdigitating microvillus lamellae from the lateral membranes and pronounced foot processes of the basal membrane that are not intertwined, but perpendicular to the basal lamina. The collecting tubule is lined by an epithelium with dark and light cells. Light cells are similar to those observed in the distal tuble except with less mitochondria and microvillus lamellae of the lateral and basal plasma membrane. Dark cells possess dark euchromatic nuclei and are filled with numerous small mitochondria. The epithelium of the neck segment, pleuroperitoneal funnel, and intermediate segment is composed entirely of ciliated cells with cilia protruding from only the central portion of the apical plasma membrane. The collecting duct is lined by a highly secretory epithelium that produces numerous membrane bound granules that stain positively for neutral carbohydrates and proteins. Apically positioned ciliated cells are intercalated between secretory cells. The collecting ducts anastomose caudally and unite with the Wolffian duct via a common collecting duct. The Wolffian duct is secretory, but not to the extent of the collecting duct, synthesizes neutral carbohydrates and proteins, and is also lined by apical ciliated cells intercalated between secretory cells. Although functional aspects associated with the morphological variation along the length of the proximal portions of the nephron have been investigated, the role of a highly secretory collecting duct has not. Historical data that implicated secretory activity concordant with mating activity, and similarity of structure and chemistry to sexual segments of the kidneys in other vertebrates, lead us to believe that the collecting duct functions as a secondary sexual organ in Ambystoma maculatum.

Ret-dependent cell rearrangements in the Wolffian duct epithelium initiate ureteric bud morphogenesis.

While the genetic control of renal branching morphogenesis has been extensively described, the cellular basis of this process remains obscure. GDNF/RET signaling is required for ureter and kidney development, and cells lacking Ret are excluded from the tips of the branching ureteric bud in chimeric kidneys. Here, we find that this exclusion results from earlier Ret-dependent cell rearrangements in the caudal Wolffian duct, which generate a specialized epithelial domain that later emerges as the tip of the primary ureteric bud. By juxtaposing cells with elevated or reduced RET activity, we find that Wolffian duct cells compete, based on RET signaling levels, to contribute to this domain. At the same time, the caudal Wolffian duct transiently converts from a simple to a pseudostratified epithelium, a process that does not require Ret. Thus, both Ret-dependent cell movements and Ret-independent changes in the Wolffian duct epithelium contribute to ureteric bud formation.

Development and morphogenesis of the Wolffian/epididymal duct, more twists and turns.

The epididymis serves a critical function of preparing the male germ cells for fertilization. In order for the epididymis to carry out this role it must undergo a highly coordinated succession of molecular and morphogenic events during development. These events begin with the formation of the Wolffian or nephric duct, the embryonic precursor of the male reproductive system, and end with the three-dimensional coiled postnatal epididymis that is comprised of several distinctly functional segments. How the duct changes from a simple straight tube to a highly convoluted structure will be the focus of this article. In reviewing the literature's current understanding of epididymal morphogenesis, we will highlight some of the classic morphological studies and discuss some of the more recent genetic models that have all served to contribute to our understanding of this system. Where published information is scarce we will provide potential hypotheses that warrant further investigation and may open up new directions of exploration using the epididymis as a model for tubular morphogenesis.

Prenatal development of the bovine oviduct.

In this study the development of the bovine Fallopian tube was investigated using light microscopic methods. Formation and differentiation of the Müllerian duct were studied in mesonephroi of 16 embryos and fetuses with a crown-rump lengths (CRL) of 0.9-8.4 cm. The funnel field, the rostral beginning of the Müllerian duct was first observed at a CRL of 0.9 cm. It appears as a thickening of the mesothelium on the craniolateral side of the mesonephros. During later development the Müllerian duct emerges by caudal outgrowth from the funnel field. Formation of a common basal lamina surrounding the caudal tips of Müllerian and Wolffian ducts could be observed at all stages up to CRL of 2.7 cm. The mesothelium and the epithelium of the Wolffian duct adjacent to the Müllerian duct showed a modification of epithelium height in all examined stages. Probably the Wolffian duct influences the growth of Müllerian duct by epithelio-mesenchymal interactions. Fetuses from a CRL of 12.0 to 94.0 cm were used for investigation of the prenatal differentiation of the oviductal mucosa. Folding of the oviductal mucosa started at a CRL of 29.0 cm and continued until birth. Individual primary, secondary and tertiary folds are formed in special proliferation zones and epithelium-folding buds. The cellular differentiation of the oviductal epithelium involves the formation of ciliated and secretory cells during different times of prenatal development. Ciliogenesis was first detected at a CRL of 33.0 cm. Active secretory cells could be observed in the oviductal epithelium from a CRL of 64.0 cm onwards.

Helper function of the Wolffian ducts and role of androgens in the development of the vagina.

The development of a vagina as a separate outlet of the birth canal evolves at the transition of egg laying species to eutherian mammals. The derivation of the vagina from the Wolffian and Müllerian ducts and the contribution of the urogenital sinus are still open questions. Here experiments with the complete androgen receptor defect in the testicular feminisation (Tfm) mouse are reported which show that the vagina is formed by caudal migration of Wolffian and Müllerian ducts. The cranial ends of the Wolffian ducts successively regress while the Müllerian ducts fuse to form the vagina. Immunohistochemistry of the androgen receptor reveals that the caudal ends of the Wolffian ducts remain in the indifferent stage and therefore have been mistaken as sinuvaginal bulbs. The Wolffian ducts do not contribute to the vagina itself but have a helper function during downward movement of the vaginal bud in the female. In the male the caudal ends serve as androgen operated switch for the negative control of vaginal development. The results indicate that the rudimentary vagina in the complete androgen insensitivity syndrome (CAIS) corresponds to non obliterated caudal ends of the Müllerian ducts. Selective atresia of the vagina in the MRKH (Mayer-Rokitansky-Kuster-Hauser) syndrome may be explained by the failure of Wolffian and Müllerian ducts to descend caudally.

Experimental contributions to the study of the embryology of the vagina.

BACKGROUND: Acién's hypothesis, deduced from patients with malformations of the female genital tract, especially those with renal agenesis and ipsilateral blind hemivagina, affirms the embryology of the human vagina as deriving from the Wolffian ducts and the Müllerian tubercle and could explain the embryological origin of all the female genital malformations reported. In this study, we investigated the hypothesis in rats. METHODS: Twenty-five pregnant rats were used to analyse female embryos (64) from day 15 (stage indifferent) to day 20 postcoitum (vagina completely formed). We performed transverse and longitudinal sections of embryos, haematoxylin-eosin tinction and immunohistochemical staining using markers specific to Wolffian derivatives. We also analysed the presence of these markers in the vagina of four adult rats. RESULTS: The Müller ducts converge until they fuse into one tube, but caudally they diverge and finally they fuse with the 'urogenital sinus bulbs' that are actually the distal portion of the Wolffian ducts according to the immunohistochemical marking with GZ1 and GZ2. The Müllerian tubercle is observed between those elements. Then, the immunohistochemical staining can be seen all along the completely formed vagina, which is also observed in the vagina of the adult rat. CONCLUSION: We prove the participation of Müller tubercle and Wolffian ducts in the formation of the vagina in rats, so we confirm experimentally Acién's hypothesis about the human vagina embryology.

Androgens and masculinization of genitalia in the spotted hyaena (Crocuta crocuta). 1. Urogenital morphology and placental androgen production during fetal life.

According to common understanding of sexual differentiation, the formation and development of a penile clitoris in female spotted hyaenas requires the presence of naturally circulating androgens during fetal life. The purpose of the present study was to determine potential source(s) of such fetal androgens by investigating the timing of urogenital development and placental production of androgen during early and mid-gestation. Fetuses determined to be female by molecular techniques (lack of SRY gene) at days 33 and 48 of gestation had undifferentiated gonads, but the clitoris was already 'masculinized' and was generally similar to the phallus of a 50-day-old male fetus. Wolffian and Müllerian ducts terminated at the urogenital sinus in both sexes and a urethra was present along the entire length of the clitoris and penis. The adrenal gland was large and histologically differentiated at 33 days. Steroid gradients across the uterus (a drop in delta 4-androstenedione, with increases in oestrogen and androgen), and high androstenedione in ovarian veins indicated that ovarian androstenedione was metabolized and secreted as testosterone by the placenta throughout gestation. In vitro, whole or homogenized placentae at days 48 and 58 of gestation (110 days total) metabolized radiolabelled androstenedione into testosterone and oestradiol; the specific enzymatic activity of early placental tissues was higher than at later stages. A human placental homogenate had higher aromatase activity but did not produce testosterone unless aromatase was inhibited. Infusion of labelled androstenedione into the uterine arteries of hyaenas demonstrated the conversion of this substrate into testosterone and oestradiol and their secretion into the fetal circulation. Evidently, androgen is produced by the placenta and secreted into the fetal circulation from early in pregnancy when masculinization is first evident, before differentiation of the fetal ovary.

Interactions of gonadal steroids and pesticides (DDT, DDE) on gonaduct growth in larval tiger salamanders, Ambystoma tigrinum.

In view of the current worldwide decline in amphibian populations, exploratory studies are needed to assess the potential for environmental contaminants to act as endocrine disrupters of the amphibian reproductive system. The present study investigated the effects of DDT dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethylene (DDE) on the development of amphibian gonaducts. Larval male and female tiger salamanders (Ambystoma tigrinum), with immature gonads, were immersed in a sublethal solution of p,p'-DDE or technical-grade DDT (80% p,p'-DDT and 20% o,p'-DDT). Additionally, larvae were injected with the steroid hormones estradiol or dihydrotestosterone (DHT). Morphometrics were used to analyze the effects and interactions of steroid and pesticide treatments on larval gonaducts. Estradiol and DHT stimulated cell proliferation and hypertrophy of the müllerian duct epithelium in both sexes. Wolffian duct epithelium, however, was stimulated only by DHT treatment. The pesticide DDT antagonized the estrogenic actions of the steroid treatments, and p,p'-DDE acted as an estrogen on the müllerian ducts of females only. The müllerian ducts of males, and the wolffian ducts of both sexes, were unaffected by DDT or DDE alone. While confirming the previously reported estrogenic actions of estradiol and DHT on urodelean gonaducts, the results contradict the expected estrogenic actions of DDT and antiandrogenic actions of p,p'-DDE. Instead, in A. tigrinum, technical-grade DDT had an antiestrogenic action and p,p'-DDE an estrogenic action.

Sexual differentiation of the urogenital system of the fetal and neonatal tammar wallaby, Macropus eugenii.

In male tammar wallabies, the scrotum is the first organ to become sexually differentiated, 4-5 days before birth (day 22 of gestation). This is followed by enlargement of the gubernaculum and processus vaginalis one day before birth. However the indifferent gonad does not show any signs of testicular cord formation or androgen production until later, at around the time of birth; this is more pronounced at 2 days post-partum (p.p.), when the testis takes on a characteristic rounded appearance. Primordial germ cells proliferate throughout the testis at this time, although the testis does not become significantly heavier than the ovary until around 80 days p.p.. In females, the appearance of the mammary glands is the first sign of sexual differentiation 4-5 days before birth. The indifferent gonad first shows signs of developing an ovarian cortex and medulla 7 days after birth. The migrating germ cells are confined to the cortex, and first start to enter meiosis about 25 days after birth. The Wolffian (mesonephric) ducts are patent to the urogenital sinus in fetuses at day 21 of gestation. In the female they have started to regress by 10 days p.p. and only rudiments remain by day 25 p.p.. The Müllerian (paramesonephric) ducts develop adjacent to the cranial pole of the mesonephros at about day 25 of gestation and grow caudally to meet the urogenital sinus between days 2 and 7 p.p.. The Müllerian duct of the female develops a prominent ostium abdominale by day 9 p.p., but this structure has completely regressed in males by day 13 p.p.. The testis and ovary both migrate caudally, together with the adjacent mesonephros, at about day 10 p.p.. The ovaries remain around the level of lumbar vertebra 4 after about day 7 p.p., while the testes continue to descend. The testes enter the internal inguinal ring at about day 25 p.p., about the time that prostatic buds first appear in the urogenital sinus, and are in the inguinal canal from days 25 to 36 p.p.. They enter the scrotum at around day 36 p.p., and testicular descent is complete by days 65-72 p.p.. Melanin develops in the tunica vaginalis 72 days after birth. The overall development of the urogenital system in this marsupial is similar to that of eutherians but the sequence of events differs, with some aspects of genital differentiation preceding gonadal differentiation, apparently because they are directly controlled by X-linked genes, rather than indirectly controlled by gonadal steroids.

Origin of müllerian duct and its later developmental changes in relation to wolffian duct in bovine fetuses.

At the 1.6-1.8 cm stages in both male and female bovine fetuses, the anlage of the Müllerian duct was first observed as an elevation of mesothelium on the cranial tip of the mesonephros. The duct extended toward the urogenital sinus in parallel with the Wolffian duct. Its caudal end was solid and was fused with the Wolffian duct at about the level of the middle of the gonadal anlage. Thereafter, the Müllerian duct became gradually independent of the Wolffian duct throughout its whole length. Therefore, it may be that the main part of the Müllerian duct is established by budding from the Wolffian duct. At the 4.3-4.5 cm stages the male Müllerian duct began to degenerate and the female one continued to grow. This suggests that the male Müllerian duct is affected by the Müllerian inhibiting substance at these stages. At the 12.0 cm stage, the female Wolffian duct disappeared. The male Wolffian duct was also much reduced in diameter at this stage, but began to grow again from the 14 cm stage onward.

Differentiation and development of the reproductive system in the iguanid lizard, Sceloporus undulatus.

Embryos of the lizard Sceloporus undulatus were sampled throughout incubation, and the differentiation and development of the reproductive system was documented histologically. The undifferentiated gonads possess both a cortex and medulla, both of which contain germ cells until embryonic stage 34. Beginning at stage 34, the cortex of the presumptive ovary thickens, and cortical germ cells are more abundant. By the time of hatching, the ovarian cortex is 6 to 10 cells thick and filled with oogonia and oocytes; primordial follicles, however, are not yet present. In males at embryonic stage 34, seminiferous tubules appear in the medulla of the testis, and Sertoli cells begin to differentiate. Seminiferous tubule formation is complete by hatching, and both Sertoli and Leydig cells are apparent. The mullerian ducts develop in both sexes but begin regressing in the male at embryonic stage 37. The wolffian ducts also develop in both sexes and are present in males and females at hatching.

The mesonephric (wolffian) and paramesonephric (müllerian) ducts of golden hamsters express different intermediate-filament proteins during development.

We analysed the expression of intermediate-filament proteins in the developing mesonephric duct (the precursor of the male genital ducts) and the paramesonephric duct (the precursor of the female genital ducts) of golden-hamster embryos using immunohistochemical methods. Embryos were investigated from the early stages of duct development, i.e. at 9.5 days post conceptionem (dpc), through sexual differentiation, until birth (15.5 dpc). Monospecific antibodies to vimentin or keratins 7, 8, 18 or 19 as well as two keratin antibodies that are pan-epithelial in human tissues were tested. Both ducts expressed vimentin to some degree from their early stages (mesonephric duct from 9.5 dpc onwards; paramesonephric duct from 10.5 dpc onwards) until birth. No keratins were detectable at these earliest stages. In the mesonephric duct, keratins 7, 18 and 19 appeared simultaneously at 10.5 dpc and persisted until birth. In the paramesonephric duct, only keratin 18 was detectable at first (at 12.0 dpc), with the expression of keratins 7 and 19 being delayed until 14.5 dpc. This feature was irrespective of sexual differentiation, which begins at 11.0 dpc, so that, in males, these keratins appeared on cue, even though the paramesonephric duct was regressing at this time. The expression of keratin 8 could not be demonstrated in either duct using the antibodies tested in our study. By 14.5 dpc, the differentiated male mesonephric duct and the differentiated female paramesonephric duct exhibited the same intermediate-filament protein pattern (weak vimentin expression and strong expression of keratins 7, 18 and 19), in spite of differences in the intermediate-filament protein patterns exhibited by the two ducts during early development. These different programmes of intermediate-filament protein regulation do not support the concept that the mesonephric duct makes a cellular contribution to the paramesonephric duct during the development of the latter.

The vagina is formed by downgrowth of Wolffian and Müllerian ducts. Graphical reconstructions from normal and Tfm mouse embryos.

We use the Tfm (testicular feminization) mutation of the mouse to reexamine the role of Wolffian and Müllerian ducts during formation of the vagina. Three dimensional graphical reconstructions of the lower genital tract are prepared from serial sections of male, female, and Tfm embryos from day 15 p.c. until 8 days after birth. The reconstructions show that in female and Tfm animals the caudal segments of Wolffian and Müllerian ducts fuse and migrate caudally, whereas in the male they do not fuse and remain in their original position. Following down-growth, separate Wolffian and Müllerian ducts emerge from the fused caudal tips of the ducts. The Wolffian ducts degenerate, while the Müllerian ducts fuse with each other and form the vagina. Wolffian and Müllerian ducts are connected to the urogenital sinus by the sinus ridges which in later stages are separated from the sinus by lateral furrows. The sinus ridges are replaced by the Müllerian ducts. We conclude that the vagina develops by down-growth of Wolffian and Müllerian ducts along the sinus ridges. Wolffian ducts and sinus ridges regress so that the definitive vagina is formed by the Müllerian ducts. In Tfm embryos the vagina forms as in the female but subsequently degenerates, probably due to the action of AMH. The vaginal pocket in the Tfm is the variable remainder of the vagina at the end of the degeneration process.