Desert hedgehog (Dhh) Gene Is Required in the Mouse Testis for Formation of Adult-Type Leydig Cells and Normal Development of Peritubular Cells and Seminiferous Tubules

An H1 histone variant, H1a, has been isolated and purified from the mouse testis. Biochemical and amino acid analyses indicate its similarity with the rat testis H1a. Specific antibodies against the purified H1a have been generated in rabbits and used to study its tissue and species distribution using protein blotting procedures. We have also used the immunocytochemical technique to determine in situ distribution of H1a in spermatogenic cells and somatic tissues of the mouse. A non-random distribution of H1a has been noted in the nuclei of certain somatic cell types such as Sertoli cells, Leydig cells and neurons. By contrast, hepatocyte nuclei lacked detectable levels of H1a. In adult seminiferous tubules, the early primary spermatocyte nuclei displayed a greater level of immunoreactivity relative to other cell types. Developmental studies indicate its initial expression in the 7-day-old mouse testis concomitant with the appearance of intermediate and type B spermatogonia.

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Expression of prostaglandin D synthetase during development in the mouse testis

Reproduction ◽

10.1530/rep.0.1220553 ◽

2001 ◽

pp. 553-559 ◽

Cited By ~ 51

Author(s):

PJ Baker ◽

PJ O'Shaughnessy

Keyword(s):

Leydig Cells ◽

Developmental Process ◽

Major Change ◽

Mouse Testis ◽

Seminiferous Tubules ◽

Interstitial Tissue ◽

Rt Pcr ◽

Subsequent Increase ◽

Splice Isoforms ◽

Neonatal Testis

Prostaglandin D synthetase is expressed relatively highly in the testis and reproductive tract of a number of species, including the mouse. In adult mouse testis, expression is confined largely to the Leydig cells and in this study changes in the expression and localization of prostaglandin D synthetase mRNA during testis development were examined. Initial studies using RT-PCR and isolated testicular compartments indicated that prostaglandin D synthetase expression in the neonatal testis was predominantly within the seminiferous tubules. In situ hybridization studies confirmed that prostaglandin D synthetase mRNA appears to be expressed only in the tubules of neonatal mouse testes and only in the interstitial tissue of the adult testis. TaqMan real-time PCR was used to quantify prostaglandin D synthetase mRNA content during development using an exogenous mRNA as a control standard. Expression per testis decreased after birth to < 10% at day 15 before recovering again by days 25-30. After day 30, expression per testis increased 40-fold during final development to adulthood. Studies using RT-PCR showed that early expression before day 15 was restricted to the tubular compartment, whereas the subsequent increase in expression after day 30 was restricted to the interstitial compartment. Database analysis showed that the 3' end of the prostaglandin D synthetase transcript was subject to alternate splicing. Both splice isoforms were shown by RT-PCR to be present throughout development and without a major change in expression pattern. These results indicate that expression of prostaglandin D synthetase mRNA shifts during development from the tubular compartment of the fetal or neonatal testis to the developing adult Leydig cells, with expression in the Leydig cells increasing markedly after puberty. These changes are similar to those observed for 17beta-hydroxysteroid dehydrogenase type III and may indicate that this developmental process is not uncommon in the testis.

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Localization of 17β-Hydroxysteroid Dehydrogenase/17-Ketosteroid Reductase Isoform Expression in the Developing Mouse Testis—Androstenedione Is the Major Androgen Secreted by Fetal/Neonatal Leydig Cells*

Endocrinology ◽

10.1210/endo.141.7.7545 ◽

2000 ◽

Vol 141 (7) ◽

pp. 2631-2637 ◽

Cited By ~ 62

Author(s):

P. J. O’Shaughnessy ◽

P. J. Baker ◽

M. Heikkilä ◽

S. Vainio ◽

A. P. McMahon

Keyword(s):

Enzyme Activity ◽

Leydig Cells ◽

Hydroxysteroid Dehydrogenase ◽

Mouse Testis ◽

Seminiferous Tubules ◽

Interstitial Tissue ◽

Androgen Synthesis ◽

Isoform Expression ◽

Neonatal Testis ◽

Type 3

The final step in the biosynthesis of testosterone is reduction of androstenedione by the enzyme 17β-hydroxysteroid dehydrogenase/17-ketosteroid reductase (17βHSD/17KSR). In this study, we have examined expression of the four known reductive isoforms of 17βHSD/17KSR (types 1, 3, 5, and 7) in the developing mouse testis and have determined changes in the localization of isoform expression and testosterone secretion during development. Using RT-PCR isoforms 1, 3, and 7 were shown to be expressed in the seminiferous tubules of neonatal testis, whereas isoforms 3 and 7 were expressed in the interstitial tissue of the adult testis. The type 7 isoform is unlikely to be involved in androgen synthesis and further study concentrated on the type 3 isoform. Developmentally, isoform type 3 was expressed in the seminiferous tubules up to day 10, showed little or no expression on day 20 and from day 30 was confined to the interstitial tissue. In situ hybridization confirmed that the type 3 isoform was expressed only in the seminiferous tubules in fetal testes and in the interstitial tissue in adult testes. In accordance with the localization of enzyme messenger RNA expression 17-ketosteroid reductase enzyme activity was very low in isolated interstitial tissue from neonatal testes while interstitial tissue from adult testes showed high activity. Seminiferous tubules from both neonatal and adult testes showed high levels of enzyme activity. The major androgen secreted by the interstitial tissue of prepubertal animals was androstenedione up to day 20 while 5α-androstanediol and/or testosterone were the major androgens secreted from day 30 onwards. These results show that fetal Leydig cells do not express significant levels of a reductive isoform of 17βHSD/17KSR and that androstenedione is the major androgen secreted by these cells. Production of testosterone up until puberty is dependent upon 17βHSD/17KSR activity in the seminiferous tubules—a“ two cell” requirement for testosterone synthesis. Expression of the 17βHSD/17KSR type 3 isoform (the main reductive isoform in the testis) declines in the seminiferous tubules before puberty but then reappears in the developing adult Leydig cell population.

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Androgen Receptor Expression in Sertoli Cells as a Function of Seminiferous Tubule Maturation in the Human Cryptorchid Testis1

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.86.1.7109 ◽

2001 ◽

Vol 86 (1) ◽

pp. 413-421 ◽

Cited By ~ 2

Author(s):

Javier Regadera ◽

Francisco MartÍnez-GarcÍa ◽

Pilar González-Peramato ◽

Alvaro Serrano ◽

Manuel Nistal ◽

...

Keyword(s):

Androgen Receptor ◽

Sertoli Cells ◽

Seminiferous Tubule ◽

Staining Intensity ◽

Receptor Expression ◽

Seminiferous Tubules ◽

Adult Type ◽

Archival Collection ◽

Cryptorchid Testis ◽

Peritubular Cells

Androgen receptor (AR) immunohistochemistry was performed in an archival collection of adult human cryptorchid testes to determine whether AR cellular distribution and intensity of immunostaining were functions of the severity of cellular dysgenesis. The seminiferous tubule histology of cryptorchid testes collected from adults is marked by three specific patterns. 1) Seminiferous tubules are characterized as maintaining focal areas of germinal cell differentiation (albeit incomplete) that are interspersed with 2) tubules composed of Sertoli cells only, these latter cells being principally of the adult type, although dysgenetic and immature Sertoli cells may also be detected. 3) In contrast, there is a class of tubule that is characterized as being composed exclusively of Sertoli cells that are extremely dysgenetic in appearance. The majority of adult-type Sertoli cells found in the first types of tubules exhibited either robust or moderate AR staining intensity. Peritubular cells of these tubules also expressed a similar AR staining intensity. In contrast, in the more dysgenetic and immature type Sertoli cells found in the second type of tubules, the intensity of AR staining was significantly less, if not missing altogether. Finally, in the most dysgenetic tubules, Sertoli cell AR staining was never detected. To our knowledge, this is the first report in the literature that addresses the intensity of AR immunostaining in Sertoli cells of cryptorchid testes. The results presented herein are consistent with the interpretation that the intensity of AR staining in Sertoli cells diminishes as a function of the severity to which the cells are afflicted within a cryptorchid testis and that focal absence of AR expression in Sertoli cells correlates with a lack of local spermatogenesis in the tubules.

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Reducing endogenous estrogen during development alters hormone production by porcine Leydig cells and seminiferous tubules

Domestic Animal Endocrinology ◽

10.1016/j.domaniend.2006.11.003 ◽

2008 ◽

Vol 34 (1) ◽

pp. 100-108 ◽

Cited By ~ 5

Author(s):

Eeman E. At-Taras ◽

In Cheul Kim ◽

Trish Berger ◽

Alan Conley ◽

Janet F. Roser

Keyword(s):

Leydig Cells ◽

Seminiferous Tubules ◽

Hormone Production ◽

Endogenous Estrogen

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Implications of body condition and seasonality on morphological and functional parameters of testes of Myotis nigricans (Chiroptera: Vespertilionidae)

Reproduction Fertility and Development ◽

10.1071/rd17316 ◽

2018 ◽

Vol 30 (7) ◽

pp. 1029

Author(s):

Marcelo Ferreira ◽

Aline Soldati ◽

Sirlene S. S. Rodrigues ◽

Laércio dos Anjos Benjamin

Keyword(s):

Light Microscopy ◽

Body Condition ◽

Environmental Variables ◽

Reproductive Strategies ◽

Leydig Cells ◽

Dry Season ◽

The Body ◽

Reproductive Capacity ◽

Seminiferous Tubules ◽

The Mean

The insectivorous bat Myotis nigricans is widely distributed throughout the Neotropics, including Brazil, and has a reproductive biology that is affected by climate and food availability. To evaluate the reproductive capacity of this species, morphofunctional parameters of the testes were correlated with environmental variables and the body condition of individuals captured. After bats had been killed, their testes were removed, fixed in Karnovsky’s fluid for 24 h and embedded in resin for evaluation by light microscopy. The mean annual tubulosomatic index (0.58%) and the percentage of seminiferous tubules in the testes (88.96%) were the highest ever recorded for the Order Chiroptera. The percentage of Leydig cells and volume of the cytoplasm of Leydig cells were higher in the rainy than dry season (80.62 ± 3.19% and 573.57 ± 166.95 μm, respectively; mean ± s.d.). Conversely, the percentage of nuclei of the Leydig cells in the dry season (26.17 ± 3.70%; mean ± s.d.) and the total number of Leydig cells (6.38 ± 1.84 × 109; mean ± s.d.) were higher in the dry season. The results of the present study could help in future conservation of these bats because they provide a better understanding of the bats’ reproductive strategies and how the species can adapt to changes.

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Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

Journal of Histochemistry & Cytochemistry ◽

10.1177/42.9.8064134 ◽

1994 ◽

Vol 42 (9) ◽

pp. 1271-1276 ◽

Cited By ~ 14

Author(s):

M Numata ◽

T Ono ◽

S Iseki

Keyword(s):

In Situ Hybridization ◽

Significant Role ◽

Meiotic Prophase ◽

Oligonucleotide Probe ◽

Mouse Testis ◽

Seminiferous Tubules ◽

Hybridization Histochemistry ◽

In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

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Sex chimaerism, fertility and sex determination in the mouse

Development ◽

10.1242/dev.113.1.311 ◽

1991 ◽

Vol 113 (1) ◽

pp. 311-325 ◽

Cited By ~ 4

Author(s):

C.E. Patek ◽

J.B. Kerr ◽

R.G. Gosden ◽

K.W. Jones ◽

K. Hardy ◽

...

Keyword(s):

Sex Determination ◽

Sertoli Cells ◽

Leydig Cells ◽

Sex Chromosome ◽

Tunica Albuginea ◽

In Situ Hybridisation ◽

Ovarian Surface Epithelium ◽

Surface Epithelium ◽

Seminiferous Tubules ◽

Coelomic Epithelium

Adult intraspecific mouse chimaeras, derived by introducing male embryonal stem cells into unsexed host blastocysts, were examined to determine whether gonadal sex was correlated with the sex chromosome composition of particular cell lineages. The fertility of XX in equilibrium XY and XY in equilibrium XY male chimaeras was also compared. The distribution of XX and XY cells in 34 XX in equilibrium XY ovaries, testes and ovotestes was determined by in situ hybridisation using a Y-chromosome-specific probe. Both XX and XY cells were found in all gonadal somatic tissues but Sertoli cells were predominantly XY and granulosa cells predominantly XX. The sex chromosome composition of the tunica albuginea and testicular surface epithelium could not, in general, be fully resolved, owing to diminished hybridisation efficiency in these tissues, but the ovarian surface epithelium (which like the testicular surface epithelium derives from the coelomic epithelium) was predominantly XX. These findings show that the claim that Sertoli cells were exclusively XY, on which some previous models of gonadal sex determination were based, was incorrect, and indicate instead that in the mechanism of Sertoli cell determination there is a step in which XX cells can be recruited. However, it remains to be established whether the sex chromosome constitution of the coelomic epithelium lineage plays a causal role in gonadal sex determination. Male chimaeras with XX in equilibrium XY testes were either sterile or less fertile than chimaeras with testes composed entirely of XY cells. This impaired fertility was associated with the loss of XY germ cells in atrophic seminiferous tubules. Since this progressive lesion was correlated with a high proportion of XX Leydig cells, we suggest that XX Leydig cells are functionally defective, and unable to support spermatogenesis.

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HYDROXYSTEROID DEHYDROGENASES IN NORMAL AND ABNORMAL HUMAN TESTES

Journal of Endocrinology ◽

10.1677/joe.0.0350239 ◽

1966 ◽

Vol 35 (3) ◽

pp. 239-NP ◽

Cited By ~ 24

Author(s):

A. H. BAILLIE ◽

W. S. MACK

Keyword(s):

Similar Pattern ◽

Leydig Cells ◽

Seminiferous Tubules ◽

Klinefelter’S Syndrome ◽

Klinefelter's Syndrome ◽

Colour Reaction ◽

Adult Human ◽

Hydroxysteroid Dehydrogenases

SUMMARY 3α-, 3β-, 11β-, 16β-, 17β- and 20β-hydroxysteroid dehydrogenases have been localized histochemically in the Leydig cells of prepubertal and adult human testes; 3α-, 16β- and 17β-hydroxysteroid dehydrogenases were present in the seminiferous tubules also. A similar pattern was found in cryptorchid testes. In addition 3β-sulphoxy steroids, including DHA sulphate, gave a good colour reaction in human Leydig cells. Testes from oestrogen-treated subjects had no histochemically demonstrable hydroxysteroid dehydrogenases and this applied also to infarcted testes. Testes from a case of Klinefelter's syndrome were found to lack 17β- and 20β-hydroxysteroid dehydrogenases in the Leydig cells. The biochemical significance of these results is discussed.

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A translational cellular model for the study of peritubular cells of the testis

Reproduction ◽

10.1530/rep-20-0100 ◽

2020 ◽

Vol 160 (2) ◽

pp. 259-268 ◽

Cited By ~ 2

Author(s):

Nina Schmid ◽

Annika Missel ◽

Stoyan Petkov ◽

Jan B Stöckl ◽

Florian Flenkenthaler ◽

...

Keyword(s):

Smooth Muscle ◽

Replicative Senescence ◽

Smooth Muscle Actin ◽

Proteome Analysis ◽

Inflammatory Factors ◽

Seminiferous Tubules ◽

Cellular Model ◽

Mechanistic Studies ◽

Peritubular Cells

Testicular peritubular cells (TPCs) are smooth muscle-like cells, which form a compartment surrounding the seminiferous tubules. Previous studies employing isolated human testicular peritubular cells (HTPCs) indicated that their roles in the testis go beyond sperm transport and include paracrine and immunological contributions. Peritubular cells from a non-human primate (MKTPCs), the common marmoset monkey, Callithrix jacchus, share a high degree of homology with HTPCs. However, like their human counterparts these cells age in vitro and replicative senescence limits in-depth functional or mechanistic studies. Therefore, a stable cellular model was established. MKTPCs of a young adult animal were immortalized by piggyBac transposition of human telomerase (hTERT), that is, without the expression of viral oncogenes. Immortalized MKTPCs (iMKTPCs) grew without discernable changes for more than 50 passages. An initial characterization revealed typical genes expressed by peritubular cells (androgen receptor (AR), smooth-muscle actin (ACTA2), calponin (CNN1)). A proteome analysis of the primary MKTPCs and the derived immortalized cell line confirmed that the cells almost completely retained their phenotype. To test whether they respond in a similar way as HTPCs, iMKTPCs were challenged with forskolin (FSK) and ATP. As HTPCs, they showed increased expression level of the StAR protein (StAR) after FSK stimulation, indicating steroidogenic capacity. ATP increased the expression of pro-inflammatory factors (e.g. IL1B; CCL7), as it is the case in HTPCs. Finally, we confirmed that iMKTPCs can efficiently be transfected. Therefore, they represent a highly relevant translational model, which allows mechanistic studies for further exploration of the roles of testicular peritubular cells.

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