Hormonal Regulation Of Spermatogenesis Research Articles

Advances in research of TGF-Β1 in human testis

Transforming growth factor β1 (TGF-β1) is a polypeptide growth factor with various biological activities. Recent studies have shown that this growth factor and they receptors are expressed in human Leydig cells, Sertoli cells, spermatogonia, spermatocytes and sperm cells after birth, and are involved in the endocrine regulation of spermatogenesis. However, there is no report on the expression of TGF-β in human embryonic testis. TGF-β1 is a peptide growth factor widely distributed in germ cells at various levels in the testis, regulating the function of the testes and maintaining the stability of the testis environment by acting on various cells. This review summarizes the research progress of TGF-β1 in human testis.

Glycoprotein Hormone Receptor Knockdown Leads to Reduced Reproductive Success in Male Aedes aegypti.

Glycoprotein hormone receptors mediate a diverse range of physiological functions in vertebrate and invertebrate organisms. The heterodimeric glycoprotein hormone GPA2/GPB5 and its receptor LGR1, constitute a recently discovered invertebrate neuroendocrine signaling system that remains to be functionally characterized. We previously reported that LGR1 is expressed in the testes of adult Aedes aegypti mosquitoes, where its immunoreactivity is particularly regionalized. Here, we show that LGR1 immunoreactivity is associated with the centriole adjunct of spermatids and is observed transiently during spermatogenesis in mosquitoes, where it may act to mediate the regulation of flagellar development. RNA interference to downregulate LGR1 expression was accomplished by feeding mosquito larvae with bacteria that produced LGR1-specific dsRNA, which led to defects in spermatozoa, characterized with shortened flagella. LGR1 knockdown mosquitoes also retained ∼60% less spermatozoa in reproductive organs and demonstrated reduced fertility compared to controls. To date, the endocrine regulation of spermatogenesis in mosquitoes remains an understudied research area. The distribution of LGR1 and detrimental effects of its knockdown on spermatogenesis in A. aegypti indicates that this heterodimeric glycoprotein hormone signaling system contributes significantly to the regulation of male reproductive biology in this important disease-vector.

Endocrine and local signaling interact to regulate spermatogenesis in zebrafish: follicle-stimulating hormone, retinoic acid and androgens.

Retinoic acid (RA) is crucial for mammalian spermatogonia differentiation, and stimulates Stra8 expression, a gene required for meiosis. Certain fish species, including zebrafish, have lost the stra8 gene. While RA still seems important for spermatogenesis in fish, it is not known which stage(s) respond to RA or whether its effects are integrated into the endocrine regulation of spermatogenesis. In zebrafish, RA promoted spermatogonia differentiation, supported androgen-stimulated meiosis, and reduced spermatocyte and spermatid apoptosis. Follicle-stimulating hormone (Fsh) stimulated RA production. Expressing a dominant-negative RA receptor variant in germ cells clearly disturbed spermatogenesis but meiosis and spermiogenesis still took place, although sperm quality was low in 6-month-old adults. This condition also activated Leydig cells. Three months later, spermatogenesis apparently had recovered, but doubling of testis weight demonstrated hypertrophy, apoptosis/DNA damage among spermatids was high and sperm quality remained low. We conclude that RA signaling is important for zebrafish spermatogenesis but is not of crucial relevance. As Fsh stimulates androgen and RA production, germ cell-mediated, RA-dependent reduction of Leydig cell activity may form a hitherto unknown intratesticular negative-feedback loop.

Constitutively active follicle-stimulating hormone receptor enables androgen-independent spermatogenesis.

Spermatogenesis is regulated by the 2 pituitary gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH). This process is considered impossible without the absolute requirement of LH-stimulated testicular testosterone (T) production. The role of FSH remains unclear because men and mice with inactivating FSH receptor (FSHR) mutations are fertile. We revisited the role of FSH in spermatogenesis using transgenic mice expressing a constitutively strongly active FSHR mutant in a LH receptor–null (LHR-null) background. The mutant FSHR reversed the azoospermia and partially restored fertility of Lhr–/– mice. The finding was initially ascribed to the residual Leydig cell T production. However, when T action was completely blocked with the potent antiandrogen flutamide, spermatogenesis persisted. Hence, completely T-independent spermatogenesis is possible through strong FSHR activation, and the dogma of T being a sine qua non for spermatogenesis may need modification. The mechanism for the finding appeared to be that FSHR activation maintained the expression of Sertoli cell genes considered androgen dependent. The translational message of our findings is the possibility of developing a new strategy of high-dose FSH treatment for spermatogenic failure. Our findings also provide an explanation of molecular pathogenesis for Pasqualini syndrome (fertile eunuchs; LH/T deficiency with persistent spermatogenesis) and explain how the hormonal regulation of spermatogenesis has shifted from FSH to T dominance during evolution.

Regulation mechanisms of spermatogenesis in red-backed mice at different population phases

This paper gives morphological and functional analysis of the endocrine and germinal testicular sections in red-backed mice Myodes glareolus from a natural population at different population sizes. The decrease in the germinal testicular activity at the peak population is due to the reduced functional activity of the endocrine testicular section (Leydig and Sertoli cells), which leads to a decreased content of testosterone, androgen-binding protein, and inhibin that take part in the hormonal regulation of spermatogenesis. The trend towards suppression of spermatogenesis is noted to persist in animals at a low population size.

Violation of spermatogenesis and outcomesof assisted reproductive technologies in various forms of hypogonadism

This article provides an overview of domestic and foreign literature on the hormonal regulation of spermatogenesis, endocrine disorders, leading to hypogonadism, on effectiveness of assisted reproductive technology in patients with Klinefelter’s syndrome, Kallmann syndrome and syndrome of testicular feminization (androgen insensitivity syndrome).

Stage-specific gene expression during spermatogenesis in the Senegalese sole (Solea senegalensis), a fish with semi-cystic type of spermatogenesis, as assessed by laser capture microdissection and absolute quantitative PCR

Spermatogenesis is a complex process where hormonal signals regulate the interaction of different cell types in a tight spatial and temporal fashion. The Senegalese sole (Solea senegalensis) is a marine flatfish that, in contrast to many fish, exhibits a semi-cystic, asynchronous pattern of spermatogenesis progression. This pattern is characterized by the release of spermatids into the tubule lumen, where they transform into spermatozoa. In this study, we used laser capture microdissection (LCM) to isolate cells from cysts containing spermatogonia, spermatocytes, spermatids or spermatozoa in order to investigate developmental patterns of gene expression. Furthermore, we also analyzed the stage-specific expression of the same set of genes throughout spermatogenesis (early-mid, late and maturing spermatogenic stages) in tissue fragments of the Senegalese sole testis. Genes analyzed by absolute qPCR in cysts isolated by LCM and stage-specific testis samples included genes involved in steroid synthesis and action (3β-hsd, 17β-hsd, 20β-hsd, star, star-like, progesterone receptor), gonadotropin action (fshr, lhr), the kisspeptin system (kiss2, kiss2r) and other genes important for the production of mature gametes (zona pellucida 2.2, claudin and clusterin). Our results show that, in general, steroidogenesis-related genes tended to increase with spermatogenesis progression and that 3β-hsd and 20β-hsd were expressed in germ cells but 17β-hsd was not. Our results also show that fshr is expressed in most testicular cell types, including germ cells. In contrast, lhr is expressed only in late spermatogenesis and is not expressed in any of the germ cell types examined, indicating that, in contrast to fshr, lhr may be primarily expressed in non-germinal cells (e.g. Leydig cells). Furthermore, kisspeptin and its receptor were expressed in all germ cell types examined and, as expected, gamete maturation-related genes were more expressed in mature stages. These results illustrate that key factors that participate in the hormonal regulation of spermatogenesis in the Senegalese sole testis show complex cell type- and stage-specific patterns of gene expression.

RFD Award Lecture 2010.Hormonal regulation of spermatogenesis: insights from constructing genetic models

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Genetic Risk Factors in Male Infertility

The etiopathogenesis of testicular failure remains unknown in about half of the cases and is referred to as "idiopathic infertility". "Idiopathic" testicular failure is of probable genetic origin since the number of genes involved in human spermatogenesis is likely thousands and only a small proportion of them have been identified and screened in infertile men. In parallel with studies aimed to identify mutations with a clear cause-effect relationship in spermatogenesis candidate genes, there is an increasing interest towards genetic susceptibility factors to male infertility. Despite many efforts, only a few clinically relevant polymorphisms have been identified. This is mainly related to the multifactorial nature of male infertility and to the inappropriate study design of the majority of the studies. The most promising polymorphisms are in genes involved in the endocrine regulation of spermatogenesis and on the Y chromosome, the "gr/gr" deletions. Polymorphisms are generally considered as co-factors. Their final effect on testis function and fertility is probably modulated by the genetic background of each individual and/or by the presence of certain environmental factors. In this review, recent findings concerning some of the most widely studied polymorphisms and male infertility will be discussed.

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<b>Endocrine Regulation of Spermatogenesis in Teleost Fish<b/>

Use of progestins in male contraception

Hormonal male contraception aims at suppression of spermatogenesis to azoospermia or at least to severe oligoasthenozoospermia, incompatible with the ability to induce a pregnancy. The general principle of this approach is based on interference with the endocrine regulation of spermatogenesis, i.e. the suppression of gonadotropins. Since both FSH (through the Sertoli cell) and LH (through the Leydig cell and testosterone (T)) are required for normal spermatogenesis, both gonadotropins need to be suppressed as strongly as possible. In East Asian men this can be achieved with T alone (preferably in depot preparations such as T undecanoate) but only two-thirds of Caucasian men respond with adequate sperm suppression. Therefore, in Caucasian men additional substances such as GnRH antagonists or progestins are required to suppress the pituitary. Over the past 30 years many combinations of various T preparations with different progestins have been tested in clinical trials. Since self-applicable steroid combinations (e.g. oral levonorgestrel or desogestrel with transdermal T) showed low effectiveness, currently injections and implants are under clinical development. Long-acting intramuscular T esters (e.g. T undecanoate), T pellets or implants (e.g. MENT) are combined with injections of DMPA or noresthisterone enanthate or with implants containing levonorgestrel or etonogestrel. Acute side-effects of these combinations appear to be minimal and tolerable, long-term effects need to be investigated.

EST analysis of gene expression in testis of the ascidian Ciona intestinalis.

To explore the gene expression underlying spermatogenesis, a large-scale analysis has been done on the cDNAs from testis of the ascidian, Ciona intestinalis. A set of 5,461 expressed sequence tags was analyzed and grouped into 2,806 independent clusters. Approximately 30% of the clusters showed significant sequence matches to the proteins reported in DDBJ/GenBank/EMBL database including a set of proteins closely related to the gene regulation during spermatogenesis, functional and morphological changes of spermatogenic cells during spermiogenesis, and physiological functions of sperm, as well as those with housekeeping functions commonly expressed in other cells. Some clones show similarities to the proteins present in vertebrate lymphocytes, suggesting a primitive immune system in ascidians. We have also found some genes that are known to participate in hormonal regulation of spermatogenesis in vertebrates. The large majority of the genes expressed in Ciona testis show no significant matches to known proteins and the further analysis of these genes may shed new light on the molecular mechanism of spermatogenesis and sperm functions.

Reproductive system: aromatase and estrogens

The cytochrome P450 aromatase (P450arom) is the terminal enzyme responsible for the formation of estrogens from androgens. In the rat testis we have immunolocalized the P450arom not only in Leydig cells but also in germ cells and especially in elongated spermatids. Related to the stage of germ cell maturation, we have shown that the level of P450arom transcripts decreases, it is much more abundant in pachytene spermatocytes (PS) than in mature germ cells whereas the aromatase activity is two- to fourfold greater in spermatozoa when compared to younger germ cell preparations. In rat germ cells, the aromatase gene expression is not only under androgen and cyclic AMP control but also subjected to cytokine (TNFα) and growth factor (TGFβ) regulation. In the bank-vole testis we have evidenced a positive correlation between a fully developed spermatogenesis and a strong immunoreactivity for both P450arom and estrogen receptor (ERβ) not only in Sertoli cells but also in PS and round spermatids (RS). Therefore, the aromatase gene expression and its translation in a fully active protein in rodent germ cells evidence an additional site for estrogen production within the testis. Our recent data showing that human ejaculated spermatozoa expressed specific transcripts for P450arom reinforced the observations reported in germ cells of other mammalian species. Together with the widespread distribution of ERs in testicular cells these data bring enlightenment on the hormonal regulation of spermatogenesis.

Hormonal regulation of spermatogenesis in primates and man: insights for development of the male hormonal contraceptive.

A better understanding of the hormonal regulation of spermatogenesis may provide new avenues for management of human infertility, and development of safe and reversible contraceptives. Although much basic work must be conducted in lower mammals, primarily rodents, concepts must, at some stage, be tested for their relevance to humans. In this review, we will draw together existing data in humans and monkeys in formulating an overview of the hormonal regulation of spermatogenesis, focusing particularly on the pursuit of the male hormonal contraceptive. Reference to data from lower mammals will be made when these highlight interesting and relevant differences or similarities with primates. The dual functions of the adult testis; namely, the production of spermatozoa (fertility) and the secretion of testosterone (virility), are both dependent on stimulation by the pituitary gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which are secreted in response to hypothalamic gonadotropin-releasing hormone (GnRH). Testosterone is essential for promoting spermatogenesis and is secreted by the adult Leydig cell under LH stimulation, and acts via androgen receptors (ARs) on Sertoli cells, Leydig cells, and peritubular cells. That testosterone exerts its effects on somatic cells rather than germ cells was highlighted by recent germ cell transplantation studies (Johnston et al, 2001). FSH acts via

Hormonal regulation of spermatogenesis in the hypophysectomized rat: cell viability after hormonal replacement in adults after intermediate periods of hypophysectomy.

A quantitative analysis of germ-cell populations in normal, hypophysectomized (Hx), and Hx-hormone-treated animals was undertaken during periods of regression that were characterized as intermediate, between short-term and long-term regression of the testis. Twenty-one groups of adult rats were administered either follicle-stimulating hormone (FSH), growth hormone, thyroid-stimulating hormone (TSH), or testosterone (T) in various doses and combinations. The dosage of T administered was less than that expected to achieve maximum testis weight. Flutamide and Casodex were used to compete with androgen binding to receptors in Hx animals, as it is known that small amounts of androgen are secreted in the absence of pituitary stimulation. Follicle-stimulating hormone, T, and TSH all significantly maintained testis weight as compared with Hx controls, although FSH and T, singly or in combination, were the most effective. Contamination of the TSH preparation with trace amounts of FSH was apparently responsible for the slight maintenance of testis weight. A novel assay for determination of the numbers of viable germ cells was used in a subset of these groups to determine the cellular sites of FSH and T action. Numbers of type A spermatogonia were lowered after Hx and were maintained by either FSH or T or a combination of these hormones. Other phases of germ-cell development most susceptible to FSH and/or T were the successive conversions of type A spermatogonia to intermediate spermatogonia, intermediate spermatogonia to type B spermatogonia, preleptotene spermatocytes to pachytene spermatocytes, and early pachytene spermatocytes to intermediate maturity pachytene spermatocytes during early and midcycle phases of pachytene spermatocyte development. Germ-cell loss during meiosis and virtually every phase of spermatid development was largely prevented by FSH or T or a combination of these hormones. Thus, in testes in advanced stages of regression, both FSH and T were capable of preventing cell loss, suggesting that both hormones can affect the survival of the same cell type. The present study demonstrated that FSH can partially compensate for lowered T levels. The combined administration of FSH and T was more effective in preventing overall cell degeneration than either hormone alone. Unlike the initial phase of spermatogenesis, in which there is a largely midcycle loss of germ cells due to Hx, the loss of cells during testis regression is more widespread and impacts several cell types in more than one stage of the spermatogenic cycle.

Hormonal regulation of spermatogenesis in the hypophysectomized rat: FSH maintenance of cellular viability during pubertal spermatogenesis.

The potential for follicle-stimulating hormone (FSH) to promote germ-cell survival and the cellular sites of FSH action were studied using a gonadally maturing (pubertal), hypophysectomized (Hx) rat model in which residual testosterone (T) activity was blocked by injections of an androgen-receptor antagonist, flutamide. Recombinant human FSH was given to androgen-deprived and androgen-blocked male rats at 27 days of age to determine maintenance of individual germ-cell types at 35 days of age. Follicle-stimulating hormone significantly increased testis weights and tubular diameters as compared with Hx and Hx-flutamide controls, although testis weights in FSH-treated animals were significantly lower than in pituitary-intact animals. Morphometric assays to determine ratios of germ cells to Sertoli cells and to determine the number of germ cells present per hour of development showed that the population of type A spermatogonia in the early stages of the cycle was not responsive to FSH. Follicle-stimulating hormone had a marked ability to maintain cell viability in the rapid, successive divisions that begin in the latter part of the cycle and that continue through the next cycle (i.e., from type A1 to A4 and from intermediate spermatogonia to type B spermatogonia to preleptotene spermatocytes to leptotene/zygotene spermatocytes to young pachytene spermatocytes). The data also suggest T responsiveness of these cell types since the Hx-FSH-flutamide group showed lower cell viability at the aforementioned steps when compared with the Hx-FSH group. Too few cell types were present at subsequent phases of spermatogenesis to allow a sensitive determination of FSH activity in the maintenance of cell viability. The data show the potential of FSH in the absence or relative absence of T activity to maintain cell viability. These data support the concept of overlapping and synergistic (or additive) effects of T and FSH in the immature rat and identify the cellular sites of FSH action.

The endocrine regulation of spermatogenesis: independent roles for testosterone and FSH

The endocrine regulation of spermatogenesis: independent roles for testosterone and FSH

Factors in nonuniform induction of azoospermia by testosterone enanthate in normal men

To identify factors differentiating men becoming azoospermic from those remaining oligozoospermic within 6 months of T treatment. Prospective, open, noncomparative contraceptive efficacy study. International multicenter study of 271 men in 10 centers in seven countries. Data from 157 achieving azoospermia and 68 remaining oligozoospermic after 6 months of treatment were analyzed. The remaining 46 men were excluded as having unclassifiable suppression status due to discontinuation before completion of suppression. Weekly IM injections of 200 mg T enanthate. Anthropometric, seminal, hormonal, and biochemical data obtained before, during, and after treatment as potential predictors of consistent azoospermia. Azoospermic men had [1] faster rates of fall in sperm output and, after a delay of 75 +/- 4 days (mean +/- SE) for sperm to reappear in the ejaculate, exhibited a faster rate of recovery of sperm output; [2] higher pretreatment levels of FSH (mean +/- SE; 3.7 +/- 0.3 versus 2.7 +/- 0.4 mIU/mL [conversion factor to SI units, 1.00]); and [3] (if treated for > 15 months) a prolonged after treatment rebound in gonadotropins compared with nonazoospermic men. There were no other differences in pretreatment variables or plasma T levels and changes in androgen-sensitive markers during treatment. None of the variables explained the higher rates of azoospermia among men in Chinese (91%, n = 3) compared with non-Chinese centers (60%, n = 7). Nonuniformity of T-induced azoospermia among healthy fertile men is not due to anthropometric or ethnic differences, to variations in androgen effects, or to poor compliance with treatment. The heterogeneity in individual susceptibility to T-induced azoospermia is most consistent with quantitative differences in the hormonal regulation of spermatogenesis and is likely to be evident with other hormonal methods for male contraception.

FSH stimulation of the production of pyruvate and lactate by rat Sertoli cells may be involved in hormonal regulation of spermatogenesis.

Summary. Sertoli cells were obtained from 3\p=n-\6-week-oldrats, which were sterile after prenatal irradiation. The production of lactate by these Sertoli cells, measured 24\p=n-\48h after isolation during incubation in the absence of hormones, increased with age of the rats from 3 to 6 weeks. At all ages investigated, the production of lactate was enhanced in the presence of FSH plus testosterone, but the stimulation was most pronounced at 4 weeks of age. Lactate production was increased by FSH alone but testosterone had no effect in the presence or absence of FSH. Sertoli cells from 4-week-old rats produced both pyruvate and lactate, which accumulated in the incubation medium in a ratio of 1:4. The stimulation of the production of pyruvate and lactate by FSH was dose\x=req- dependent (ED50 at ~10 ng NIH-FSH-S13/ml). The production of pyruvate and lactate was stimulated 2-fold by insulin, 4-fold by FSH and > 6-fold by dibutyryl cyclic AMP (in the presence of 3-isobutyl-1-methylxanthine). The effects of FSH (0\m=.\5\g=m\g NIH-FSH-S13/ml) and insulin (5 \g=m\g/ml) were not additive. Leucine incorporation into isolated pachytene spermatocytes and round spermatids was stimulated by exogenous pyruvate and lactate in a dose-dependent way : maximal incorporation was obtained with 0\m=.\2 mM-pyruvate or 2 mM L-lactate. Spent medium from incubated Sertoli cells (from 4-week-old rats) stimulated the leucine incorporation into isolated pachytene spermatocytes and round spermatids 4\p=n-\8-fold.This effect could be explained by the amounts of pyruvate and lactate present in the spent medium. It is suggested that pyruvate and lactate are major products from Sertoli cells which can support synthetic activities in germ cells, and the present results indicate that pyruvate and lactate may play a role in the hormonal regulation of spermatogenesis.

Endocrine regulation of spermatogenesis in Nereis diversicolor (annelida polychaeta): experimental study of the control of meiotic differentiation.

In Nereidae, spermatogenesis is regulated by a hormone secreted by the supraesophageal ganglion (brain hormone). The hormonal concentration decreases with worm age. Spermatogonial proliferation in young worms proceeds under a high hormonal level whereas differentiation of spermatozoa in aged worms occurs in the absence of brain hormone. Spermatogonia were removed from the endocrine influence of the brain in vitro by the organ culture method. Isolated parapodia produced spermatozoa after 13 days. Control cultures were obtained by associating a parapodium with the prostomium of a young worm. It seems that the brain hormone inhibits the spermatocyte differentiation since removal of the brain results in increased RNA synthesis, followed by DNA replication, meiosis, and spermiogenesis (Bertout, '83). Modalities of the hormonal control of meiosis, especially at the level of the RNA syntheses related to meiotic differentiation, were investigated. Effects of reintroducing the hormonal influence during meiotic differentiation were studied through parapodium-prostomium associations at various time after the brain inhibition has been lifted. Effects or RNA inhibitors (actinomycin D, alpha-amanitin, cordycepin) were also tested. The results lead us to propose a hypothesis according to which brain hormone would interfere with molecular events related to the transition from the stage of spermatogonial proliferation (mitotic behavior) to the stage of meiotic differentiation (meiotic behavior).