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Expression of anti-Müllerian hormone mRNA during gonadal and follicular development in the brushtail possum (Trichosurus vulpecula).

The ontogeny of anti-Müllerian hormone (AMH) gene expression in the brushtail possum during formation of the ovary and growth of follicles was examined using in situ hybridization. For comparative purposes, the expression pattern of AMH was also examined in the developing testis. In the female, AMH mRNA was observed in the ovary of 50% (3/6) of pouch young collected around the time of sexual differentiation of the gonad (Days 1-5): the signal was predominately localized to the inner-cortical and outer-medullary region of the ovary. Thereafter, AMH mRNA was not observed in the developing ovary until Days 78-113 of postnatal life when follicles first formed at the cortical-medullary boundary. At this time, AMH mRNA was observed in the cuboidal granulosa cells of some early growing (i.e. transitional) follicles and in the granulosa cells of primary follicles. Thereafter, AMH mRNA was present in granulosa cells at all subsequent stages of follicular growth (i.e. primary through antral), but not in preovulatory follicles. In all cases, once follicles had formed, AMH mRNA was limited to the granulosa cells and was not observed in the surface epithelium, stromal cells, oocytes, theca, corpus luteum, medullary cords, rete or interstitial glands. In the possum testis, Sertoli cells strongly expressed AMH around the time of sexual differentiation of the gonad, but expression decreased to very low levels in adults, suggesting that AMH plays a similar role in brushtail possums to that observed in other mammalian species. In conclusion, localization of mRNA for AMH exclusively to granulosa cells of growing follicles in the brushtail possum is consistent with a central role for this hormone in control of granulosa cell function in marsupials. In addition, expression of AMH in the developing ovary around the time of morphological sexual differentiation raises intriguing questions regarding the possible role of AMH at this time.

Animals↗

The fate of germ cells in the testis of fetal Sex-reversed mice.

XX germ cells in the fetal testes of XX Sex-reverse male mice were observed mostly to develop in the same manner as do XY germ cells in a normal fetal testis; but some, in the vicinity of the mesonephric rete region, entered the prophase of meiosis at the same time as do XX germ cells in a normal fetal ovary. No germ cells in meiosis were found in the fetal testes of XY males, nor of XO Sex-reversed males. It is suggested that a second X chromosome renders a germ cell more susceptible to the meiosis-inducing influence of the mesonephric rete.

Animals↗

Cell-specific expression of betaC-activin in the rat reproductive tract, adrenal and liver.

betaC-activin expression was assessed in rat tissues, using reverse transcription and real-time polymerase chain reaction, Western blotting and immunohistochemistry with a specific monoclonal antibody. betaC-activin mRNA was predominantly expressed in liver, but significant amounts were found in rat whole pituitary extracts (n = 5), and in three of five extracts of ovary, testis, and adrenal gland. Specific betaC-activin immunoreactivity was demonstrated in the cytoplasm of hepatocytes, neurosecretory cell terminals in posterior pituitary, ovarian primordial follicles, theca interna, large luteal cells and rete ovarii, spermatogonia, pachytene spermatocytes and Leydig cells of the testis, uterine endometrium, oviduct epithelium and zona glomerulosa of the adrenal. The observation of stage-specific expression in gonadal cells suggests this activin subunit has specific roles, different from those of other activin/inhibin subunits. Small amounts of mRNA in the presence of significant betaC-activin protein highlights the importance of examining betaC-activin expression at both the mRNA and protein level.

Adrenal Glands↗

Blood supply to the testis of a Brazilian marsupial (Didelphis azarae) and its abdominotesticular temperature gradient.

The testicular arteries of Didelphis azarae originate from the abdominal aorta either independently from each other or by way of a common trunk. Accessory testicular arteries may be found. At the spermatic cord they form a rete mirabile having 26.8 +/- 5.0 and 29.3 +/- 4.9 slender branches on the right and left sides, respectively. The arterial branches are intermingled with veins of similar caliber and number. Near the testis the branches of the rete reunite in a single vessel which then penetrates the parenchyma of the testis. Inside the testis the artery divides usually into two main branches that course toward the caudal pole. The rectal, scrotal and testicular temperatures were 32, 28.5 and 30.4 degrees C, respectively, appearing than an abdominotesticular gradient temperature exists in this animal. Whether this mechanism is thermoregulatory for the normal spermatogenesis cannot be inferred from the present work.

Animals↗

Testicular development in Chinese Meishan boars.

The developmental process of the testis and age-related changes in the morphology of rete testicular spermatozoa were investigated in Meishan boars at 1 to 364 days of age. Testicular weight and the diameter of seminiferous tubules increased rapidly until 150 to 180 days of age. Leptotene stage spermatocytes, round spermatids and spermatozoa were first found in the section of seminiferous tubules at 30 to 45, 60 and 75 days of age, respectively. However, after 105 to 120 days of age, most rete testicular spermatozoa were morphologically normal. These results indicate that Meishan boars reach puberty as early as 75 days of age, though the testes acquire the ability to produce morphologically normal spermatozoa at about 120 days.

Journal Article↗

[Vascular morphology of the bovine testis. Light and scanning electron microscopic studies].

The testicular artery of the bull-testis shows a straight course from the end of the pampiniform plexus to the caudal extremity of testis. There it branches off in Rami tunicales, which lie as stratum arteriosum superficially to the albugineal veins of testis: a multi-layered stratum venosum. Arterial Rami parenchymales centripetales run directly to the mediastinum testis, form coils and then divide in approximately 10 or more thinner Rami parenchymales centrifugales, which extend from the coils into the parenchyma of the gonad. The three-dimensional microvasculature of the bull-testis is strikingly different from that of rodents: The peritubular network of capillaries in the interstitial space is positioned in a more irregular way. Only here and there is discernible a rope-ladder-like or polygonal arrangement of capillaries. A subalbugineal plexus does not exist in the bovine testis. Parenchymal veins drain in albugineal veins and these empty in the venous networks of the pampiniform plexus. Valves are a rare finding in testicular veins. Already low perfusion-pressure easily forces the corrosion-compound to leave the capillary bed and form typical extravasations as bent shovel-like plates, thus filling the clefts of peritubular spaces. Arteries and veins are directly embedded in the parenchyma of testis, surrounded only by a relatively thin margin of perivascular connective tissue. There are no septula testis and therefore a lobular organisation of bovine testis does not exist. The angioarchitecture of the testis plays an important role in thermoregulatory and androgen-transfer mechanisms as well as in the transport of rete-fluid to the epididymis.

Animals↗

A histologic study of the auto-allergic testis lesion in the guinea pig.

The auto-allergic lesion in guinea pigs inoculated with homologous testis plus the Freund adjuvant was investigated histologically. The lesion was found to consist of disseminated foci of perivenous inflammation, lymphocytes and histiocytes predominating in the cellular infiltrate, with invasion of epididymal, rete, and seminiferous tubules and destruction of tubular contents. Guinea pigs up to 800 gm. showed a rapidly progressing diffuse hypo- or aspermatogenesis, which appeared to be secondary to the inflammatory disease. In these animals, the process resolved leaving an atrophic testis with few or no indications of the preceding inflammation and fibrotic scarring only in the rare instances in which actual necrosis of connective tissue elements had occurred. In 1200 gm. animals there was no general hypospermatogenesis and the late findings were limited to foci of aspermatogenesis. This disease then is an experimental auto-allergic orchitis followed by testicular atrophy without scarring. Its morphologic similarity to mumps orchitis and to sterility with "germinal cell aplasia" in man is commented on.

Animals↗

Gonadal development in the opossum, Monodelphis domestica: the rete ovarii does not contribute to the steroidogenic tissues.

The gonads of 273 animals, ranging in age from newborn to adult, were examined in serial histological sections. Primordial germ cells were located in the hindgut, dorsal mesentery and gonadal primordia of neonates but were alkaline phosphatase negative. The testis differentiated between d 13 of gestation and birth, about half a day later. Testis cords, composed of pre-Sertoli cells and containing germ cells, were distributed peripherally in the gonad. Ovarian differentiation began on d 6, when an ill defined cortex and medulla became discernible. Meiosis commenced on d 14, medullary cords formed from blastema cells by d 26 and gave rise to granulosa cells around d 29. The rete ovarii was first observed in the hilar region of the gonad primordium. It penetrated maximally two thirds of the ovarian length between d 26 and d 29. It is concluded that, at least postnatally, the mesonephros does not contribute to the rete ovarii and that the granulosa cells are derived not from the rete but from the medullary cords.

Animals↗

Differential distribution of type IV collagen chains in the developing rat testis and ovary.

The localization of type IV collagen alpha 1-alpha 5 chains in the differentiating rat testis and ovary was studied by immunocytochemistry. The initial formation of the testis and ovary included the appearance of collagen alpha 1/alpha 2(IV) chains in the gonadal blastemas. Upon further differentiation of the epithelia of the gonads alpha 1/alpha 2(IV) chains became localized in all of the respective basement membranes (BMs). The alpha 3, alpha 4 and alpha 5 chains of type IV collagen were not detectable in the prenatal rat testis and ovary. With the postnatal differentiation of the rat testis the alpha 3-alpha 5(IV) chains gradually appeared, and were localized in BMs of the testicular cords and seminiferous tubules, rete cords, myoid cells, surface epithelium, Leydig cells, and in some blood vessels. In the postnatal rat ovary, the alpha 3(IV) chain appeared in the BMs of small cortical follicles whereas the BMs of secondary and more deeply localized follicles were devoid of this chain. The alpha 1/alpha 2(IV) chains were abundant in the theca. A reaction for alpha 3-alpha 5(IV) chains also appeared in the BM of the ovarian surface epithelium and of some blood vessels after birth. The present results show that the alpha 3-alpha 5(IV) chains are not only less widely distributed than the alpha 1/alpha 2(IV) chains but are also synthesized much later in development. The late appearance of the alpha 3-alpha 5(IV) chains shows that the development of the mature testicular and ovarian BMs is a long process and that the time schedule for the synthesis of these chains is different from that of many other extracellular matrix proteins. A careful analysis of the expression of alpha 3(IV) chain may be useful in the further study of the kinetics and regulation of ovarian follicular growth.

Animals↗

Immunolocalization of receptors for androgen and estrogen in male caprine reproductive tissues: unique distribution of estrogen receptors in efferent ductule epithelium.

Androgens and estrogens affect physiological processes in the testis and male excurrent duct system. This study was designed to identify and characterize distribution of androgen receptors (AR) and estrogen receptors (ER) in the reproductive organs of the male goat. Tissues, including testis, efferent ductules, epididymis (regions I-V), and ductus deferens, were obtained from five mature Nubian goats, fixed in 4% paraformaldehyde, and embedded in paraplast. Antigenic sites for AR were unmasked by microwave treatment (four times, 5 min each) of tissue sections immersed in 10 mM citrate (pH 6) and were detected using the PG-21 rabbit anti-rat/human antibody. Antigenic sites for ER were identified using the H-222 rat anti-human monoclonal antibody after tissue sections were treated with pronase (0.5 mg/ml, 37 degrees C, 8 min). Avidin-biotin horseradish peroxidase procedures were used to identify positive immunoreactivity. Irrelevant IgG was substituted for primary antibody in negative controls. Positive nuclear immunostaining for AR was observed in all types of epithelial cells, peritubular smooth muscle cells, and intertubular fibroblasts of the intratesticular rete, efferent ductules, epididymis (regions I-V), and ductus deferens, as well as in Sertoli, Leydig, and peritubular myoid cells and intertubular fibroblasts of the testis. In contrast, nuclear immunostaining for ER was confined to nonciliated cells of the efferent ductules. Thus, AR-positive cells are ubiquitously distributed in caprine testicular and excurrent ductular tissues, and ER-positive cells are unique to the efferent ductules. The caprine model should be useful in studies designed to determine mechanisms through which androgens and estrogens regulate development and function of the testes and excurrent ducts.

Animals↗

Testis differentiation in the fetal and postnatal ferret.

Testis development has been examined in a series of 59 fetal and 9 postnatal ferrets from day 22 of the 40-42 day pregnancy, to 12 days after birth. Developing seminiferous tubules and interstitial cells were first seen on day 26, and were well established one day later. Differentiation was associated with cell enlargement and cell division, and consequently the testes grew distinctly more rapidly at this time than the corresponding ovaries. Up to the end of pregnancy the epithelioid interstitial cells, derived from stromal tissue, formed a large proportion of the testis. Lipid was distinguishable in them, histologically, from about day 30. As in various other mammals, there appeared to be some regression of the interstitial Leydig cells around the time of birth, when the seminiferous tubules resumed their growth. The intra-gonadal rete was present from day 22 onwards; it established connexions with the seminiferous tubules through the small tubuli recti by day 32.

Age Factors↗

Post-hypophysectomy ovarian senescence and its relation to the spontaneous structural changes in the ovary of intact aged rats.

Hypophysectomy performed in 25- to 26-day-old Wistar rats leads within 1 years to the formation of ovarian testis-like tubes and epithelial cellular cords which are typical structures in the senile ovaries of normal 24-month-old rats. Testis-like tubes represent an unusual late stage of follicular degeneration; the origin of the cords is more complex; the revival of cell differentiation from stroma and/or rete ovarii in the absence of the pituitary is hypothesized. At 16 months of age, cords proliferate and are responsible for ovarian weight increase to almost twice the minimal weight seen 4 months after the operation. Variability in cord proliferation is considerable from one rat to another, but also between the two ovaries of the same animal. Thus, an intrinsic age-related ovarian factor is implicated in cord proliferation. From this study, it is inferred that during intact rat senescence, intrinsic ovarian aging is responsible for the proliferation of cords, whereas their induction depends on a hypothalamic-hypophyseal imbalance occurring at about 1 year of age.

Aging↗

Alterations in the immunohistochemical localization patterns of alpha-smooth muscle actin (SMA) and vimentin in the postnatally developing bovine cryptorchid testis.

Previously, we reported the normal postnatal developmental changes in immunohistochemical localization of alpha-smooth muscle actin (SMA) and vimentin in the bovine testis. In this study, we demonstrate the alterations of these cytoskeletal proteins in the bovine cryptorchid testis as compared to the contralateral scrotal testis during postnatal development. Seminiferous peritubular alpha-SMA did not appear in the cryptorchid testis until 8 months of age, except for very weak intermittent filaments in relatively larger seminiferous tubules. However, a similar peritubular pattern was observed in the 18-month-old cryptorchid and scrotal testes. Moreover, weak expression of alpha-SMA in the straight tubules and rete testes at 5 months of age did not improve until 18 months of age in the cryptorchid testes. The Sertoli cell vimentin in the cryptorchid testes revealed a highly immature pattern at 5 months of age, a pattern similar to a transforming pattern with infranuclear vimentin extensions at 8 months of age, and a pattern that was almost a transforming pattern, but with considerable weakening of the vimentin filaments, at 18 months of age. In conclusion, cryptorchidism may cause considerable delay in testicular myoid cell differentiation and in attainment of the transforming pattern of the Sertoli cell vimentin, which weakens and fails to attain the mature pattern in the cryptorchid testis. These alterations may be related to the structural immaturity and functional failure of postnatally developing bovine testes exposed continuously to body heat.

Actins↗

Differential distribution of the alpha 6 subunit of integrins in the development and sexual differentiation of the mouse testis.

The distribution of the alpha 6 subunit of integrins in the development and sexual differentiation of mouse testis was analyzed by light and electron microscopy during the embryonic, fetal and early postnatal periods. At the pregonadal phase only the epithelial cells of the mesonephric duct and of the distal mesonephric tubules showed a reaction to alpha 6, whereas the surface epithelium and the mesenchyme of the mesonephros were negative or contained only a rudimentary amount of the alpha 6 subunit. With the formation of the gonadal ridge and the testicular blastema, the gonadal cells became positive for the alpha 6 subunit. This expression remained in embryonic cord cells and in the vascular endothelial cells, whereas the differentiating cells of the surface epithelium, tunica albuginea, the Leydig cells, and the interstitial mesenchymal cells were negative. With the fetal and postnatal differentiation, the expression of the alpha 6 subunit gradually diminished in the cord cells, and by the prepubertal phase, alpha 6 was found only at adhesion sites between some Sertoli cells. Similar changes were seen in the mesonephric duct and tubules, and in the rete cords. The presence of alpha 6 in regions undergoing developmental cell aggregation processes and their disappearance during tissue maturation, suggest that alpha 6 plays a specific but transient role in gonadal cell adhesion necessary for the histogenetic organization of the testis. In addition to its role in developing and organizing cells, alpha 6 integrin was also a prominent component in degenerating cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Human testicular development and the role of the mesonephros in the origin of a dual Sertoli cell system.

Some aspects of the development of the human testis (and overy) are discussed and the main theories regarding gonadal differentiation summarized. The major part of this review deals with the origin and differentiation of the three groups of somatic cellular content: Sertoli cells, Leydig cells and peritubular cells. The most important role of the mesonephros in gonadal development is described. Under the influence of the mesonephros, a second type of meiosis-inducing Sertoli cell differentiates and becomes the opponent of a meiosis-preventing type of Sertoli cell which derives from the coelomic epithelium. All somatic cells are pooled in the central gonadal blastema which is part of the medulla. They migrate via the rete blastema to the sites of their final differentiation. Included are the precursors of the Leydig cells and the peritubular cells.

Cell Differentiation↗

Evaluation of testicular toxicology: a synopsis and discussion of the recommendations proposed by the Society of Toxicologic Pathology.

BACKGROUND: Detection of chemically induced effects on male fertility and on testicular spermatogenesis in particular, has become of increasing concern. More stringent regulatory guidelines, introduced by ICH, EPA and OECD (Table 1) have raised the awareness of toxicologists and pathologists for the need to conduct sensitive and careful evaluation of the male reproductive tract for potential toxic effects of administered compounds. With it has come confusion and in many cases, inappropriate procedures, often based on misunderstanding of what is required and on inadequate understanding of spermatogenesis. This article summarizes and discusses the main recommendations recently proposed by the Society of Toxicologic Pathology on recommended approaches for the evaluation of testicular and epididymal toxicity [Lanning LL, Creasy DM, Chapin RE, Mann PC, Barlow NJ, Regan KS, Goodman DG. Toxicologic Pathology 30:518-531, 2002]. The major recommendations are: Use sexually mature animals to evaluate effects on spermatogenesis. Sample left and right testes and epididymides and record organ weights. Use modified Davidson's fixative to fix testes from all species from studies of 13 wks duration and less. Examine transverse sections of the testes (including part of the rete), and longitudinal sections of the epididymides. Embed tissues in paraffin wax. For rodent studies up to 28 days, examine periodic acid-Schiff's-hematoxylin stained sections. For all other studies examine hematoxylin and eosin stained sections. Microscopic evaluation of the testis should be a qualitative evaluation carried out with an awareness of the spermatogenic cycle. Quantitative procedures are inappropriate for screening studies. Nomenclature and grading of findings for spermatogenic disturbances will vary on a case by case basis.

Animals↗

Effects of obstruction of the flow of seminiferous tubule fluid on the germinal epithelium in the rat.

Blocking the lumen of a single seminiferous tubule by introducing a plug of non-toxic latex produced a lesion in that tubule, but not in immediately adjacent tubules. The lesion extended for up to 50 mm from the end of the latex. Nearest to the block the tubule was completely aspermatogenic; further along the tubule, the lesion was less severe, involving disorganization and reduction in germ cell numbers, with the cells showing vacuolation, pycnosis and karyolysis. Binucleate and giant cells were common, and cells were often exfoliated into the lumen. The lesion tended to increase in length with longer times after introduction of the plug, but there appeared to be no preferential involvement of the shorter segment of the tubule between the block and the rete. The transition from damaged to healthy tubule was abrupt.

Animals↗