Germ Cell Maturation Research Articles

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Objectives: The present study was conducted to investigate the effects ofrecombinant human interferon-α-2b (rh-INF-α-2b) on testicular histomorphology in adult ratmodel. Study Design: Experimental study. Place & Duration: Animal house, Sindh AgricultureUniversity Tando Jam and Isra University Hyderabad from January to December 2014.Methodology: 80 adult albino rats were selected according to inclusion and exclusion criteriaand divided into 4 groups. Group I: Control rats 0.9% saline, Group II: rhIFN α-2b (3MIU),Group III: rhIFN α-2b (5MIU) and Group IV received rhIFN α-2b (10MIU). The rhIFN α-2b wasinjected intra-peritoneal (i.p) three times a week for 3 weeks in doses of 3MIU, 5MIU and10MIU. Animals were euthanized. Orchidectomy was performed and testicles were stored in10% formaldehyde. 5μ thick tissue sections were stained by Hematoxylin & Eosin (H & E).Results: Atrophic seminiferous tubules with clumping of lining epithelia were noted. Germ cellmaturation arrested was prominent; hypervascularity with reduced germ cells and sperm cellswere noted in high dose rhIFN α-2b treated groups. Tubular desquamation and thick basementmembrane were visible. The sertoli cells and interstitial cells of Leydig counts were increased.Conclusion: It is concluded that the recombinant human interferon-α-2b exerts serious adverseeffects on testicular histomorphology.

What Animal Models of Testicular Descent and Germ Cell Maturation Tell Us about the Mechanism in Humans.

Testicular descent occurs in most mammals in two main steps that have different hormonal control and anatomical processes. The evolution of testicular descent reveals the same basic processes in humans and animals, with minor differences in timing and anatomy, especially the location of the scrotum and the processus vaginalis. Animal models are useful as they reveal some embryological processes that cannot be studies easily in humans, such as the potential role of the mammary line and the role of the genitofemoral nerve. Postnatal germ cell development is very similar in animal models and humans, except for the timing of arrival of the testis into the scrotum, which is before birth in humans versus around puberty in rodents. Once all the minor differences between animal models and humans are taken into account, animal experimentation has provided amazing insights into the mechanisms of testicular descent, and recently, how the postnatal germ cell develops in normally descended and undescended testes.

Dearth and Delayed Maturation of Testicular Germ Cells in Fanconi Anemia E Mutant Male Mice.

After using a self-inactivating lentivirus for non-targeted insertional mutagenesis in mice, we identified a transgenic family with a recessive mutation that resulted in reduced fertility in homozygous transgenic mice. The lentiviral integration site was amplified by inverse PCR. Sequencing revealed that integration had occurred in intron 8 of the mouse Fance gene, which encodes the Fanconi anemia E (Fance) protein. Fanconi anemia (FA) proteins play pivotal roles in cellular responses to DNA damage and Fance acts as a molecular bridge between the FA core complex and Fancd2. To investigate the reduced fertility in the mutant males, we analyzed postnatal development of testicular germ cells. At one week after birth, most tubules in the mutant testes contained few or no germ cells. Over the next 2–3 weeks, germ cells accumulated in a limited number of tubules, so that some tubules contained germ cells around the full periphery of the tubule. Once sufficient numbers of germ cells had accumulated, they began to undergo the later stages of spermatogenesis. Immunoassays revealed that the Fancd2 protein accumulated around the periphery of the nucleus in normal developing spermatocytes, but we did not detect a similar localization of Fancd2 in the Fance mutant testes. Our assays indicate that although Fance mutant males are germ cell deficient at birth, the extant germ cells can proliferate and, if they reach a threshold density, can differentiate into mature sperm. Analogous to previous studies of FA genes in mice, our results show that the Fance protein plays an important, but not absolutely essential, role in the initial developmental expansion of the male germ line.

Vocal sac development and accelerated sexual maturity in the lesser swimming frog, Pseudis minuta (Anura, Hylidae)

Sexual maturity involves the differentiation of the reproductive system, the maturation of germ cells, and the development of secondary sexual characteristics. Even though this topic has received much attention, little is known about the sequence of events that encompass reproductive maturation in anurans and what it could reveal about the developmental basis of life cycle evolution. The discovery of froglets of Pseudis minuta with incipient vocal sacs calling in breeding pools alongside several larger adult specimens with fully developed vocal sacs raised the question of the timing of sexual maturity in this species. Here we describe the sequence and timing of differentiation, development and maturation of the vocal sac apparatus and the testes in P. minuta (Anura, Hylidae), in order to establish a timeline of events leading to sexual maturity. Differentiation of the vocal sac apparatus begins at the final metamorphic stages, earlier than reported for other species, and the vocal sac acquires its final shape during the early postmetamorphic period. These modifications occur after gonadal differentiation, which begins early during the larval period and proceeds with a highly accelerated rate of development (e.g., secondary spermatids appear well before metamorphic climax), a situation reported previously for other anuran species only in the genus Pseudis. These results, together with a skeletochronological analysis showing that some calling specimens presented no lines of arrested growth, indicate acceleration in the timing of sexual maturity in Pseudis, and raise questions about the interdependence/decoupling during the development of the different components involved in reaching the adult stage.

Interaction of herpesviruses with mature human spermatozoa in the model system in vitro

The DNA of human herpesviruses (HHV), including the herpes simplex virus (HSV) and cytomegalovirus (CMV), is often identified in ejaculates of patients with urogenital diseases and infertility. At least a part of viral DNA is associated with cell fraction of ejaculate. However, it remains unclear how the semen is infected by the virus. It can be located in gametes or be capable of infecting mature germ cells, including motile sperm cells. In order to resolve this issue, interactions of the CMV and HSV with human sperm cells were studied using an original optimized model of the herpesviral infection of male gametes in vitro. The analysis of the immunofluorescent staining of gametes for viral antigens has shown that CMV infected 2% gametes, while HSV infected 17.26 ± 2.58% gametes. The fraction of progressively motile sperm cells contained 13.99 ± 4.64% infected cells. Localization of HSV was studied by the confocal microscopy. Sometimes, viral gB protein was found on sperm cell membrane. In addition, optical scanning of other cells has shown the intracellular localization of the viral proteins. In the majority of spermatozoa, the viral proteins were observed in the head and neck. In some cells, they were located in the middle piece or, rarely, in the equatorial segment. In general, after in vitro infection HSV antigens were located in the same areas of the sperm cells as in ejaculates from infected patients. According to DNA-DNAhybridization in situ, gametes containingHSV DNAaccounted for 16.94 ± 5.28%, which is consistent with the results obtained in the immunofluorescence assay. It can be concluded that mature male gametes are infected by HHV in the genital tract, where the virus binds to the sperm cell membrane and enters the cell. Interaction of HHV with progressively motile sperm cells implies a vertical viral transmission upon fertilization and points to the necessity of testing ejaculate for herpesviruses infections.

Re. “Prospective study of histological and endocrine parameters of gonadal function in boys with cryptorchidism”

Verkauskasa et al. are to be commended on the depth of their investigation on hormonal parameters and histological changes in boys with cryptorchidism [1]. The overall finding of the study, which is that Ad spermatogonia counts decrease with age of cryptorchid testis biopsy, is consistent with other studies and highlights the need for prompt treatment of cryptorchid patients after 6 months of age [2]. In the subgroup that had a high risk of infertility because of absence of Ad spermatogonia stem cells, the LH levels were lower than expected, consistent with this subgroup having a hypogonadotropic hypogonadism. However, this group is small, less than 20% of the total sample. While the Ad numbers were low in the patients studied, it appears that the count was zero in only 21/51 pts (i.e. the majority of testes showed some germ cell maturation). The authors claim that transformation of gonocytes into Ad spermatogonia is testosterone dependent; however, there are also contrary animal and human studies which show that, even in complete androgen insensitivity, Ad spermatogonia appear normal initially. This would suggest that gonocyte transformation into Ad spermatogonia may be androgen-independent. Furthermore, several recent studies in animals and humans show that the pathogenesis and etiology of isolated cryptorchidism is likely multifactorial and in most cases unknown [3]. The bottom line is this: while men with a history of unilateral cryptorchidism tend to have lower sperm counts than the normal

Myt1 inhibition of Cyclin A/Cdk1 is essential for fusome integrity and premeiotic centriole engagement in Drosophila spermatocytes.

Regulation of cell cycle arrest in premeiotic G2 phase coordinates germ cell maturation and meiotic cell division with hormonal and developmental signals by mechanisms that control Cyclin B synthesis and inhibitory phosphorylation of the M-phase kinase, Cdk1. In this study, we investigated how inhibitory phosphorylation of Cdk1 by Myt1 kinase regulates premeiotic G2 phase of Drosophila male meiosis. Immature spermatocytes lacking Myt1 activity exhibit two distinct defects: disrupted intercellular bridges (fusomes) and premature centriole disengagement. As a result, the myt1 mutant spermatocytes enter meiosis with multipolar spindles. These myt1 defects can be suppressed by depletion of Cyclin A activity or ectopic expression of Wee1 (a partially redundant Cdk1 inhibitory kinase) and phenocopied by expression of a Cdk1F mutant defective for inhibitory phosphorylation. We therefore conclude that Myt1 inhibition of Cyclin A/Cdk1 is essential for normal fusome behavior and centriole engagement during premeiotic G2 arrest of Drosophila male meiosis. The novel meiotic functions we discovered for Myt1 kinase are spatially and temporally distinct from previously described functions of Myt1 as an inhibitor of Cyclin B/Cdk1 to regulate G2/MI timing.

Charactering the ZFAND3 gene mapped in the sex-determining locus in hybrid tilapia (Oreochromis spp.).

Zinc finger AN1-type domain 3 (ZFAND3) is essential for spermatogenesis in mice. However, its function in teleosts remains unclear. In this study, we characterized the ZFAND3 gene (termed as OsZFAND3) in an important food fish, tilapia. The OsZFAND3 cDNA sequence is 1,050 bp in length, containing an ORF of 615 bp, which encodes a putative peptide of 204 amino acid residues. Quantitative real-time PCR revealed that the OsZFAND3 transcripts were exclusively expressed in the testis and ovary. In situ hybridization showed that the high expression of OsZFAND3 transcripts was predominantly localized in the spermatocyte and spermatid. These results suggest that OsZFAND3 is involved in male germ cell maturation. Three single nucleotide polymorphisms (SNPs) were detected in the introns of OsZFAND3. The OsZFAND3 gene was mapped in the sex-determining locus on linkage group 1 (LG1). The three SNPs in the OsZFAND3 gene were strictly associated with sex phenotype, suggesting that the OsZFAND3 gene is tightly linked to the sex-determining locus. Our study provides new insights into the functions of the OsZFAND3 gene in tilapia and a foundation for further detailed analysis of the OsZFAND3 gene in sex determination and differentiation.

The mRNA-bound proteome of the early fly embryo.

Early embryogenesis is characterized by the maternal to zygotic transition (MZT), in which maternally deposited messenger RNAs are degraded while zygotic transcription begins. Before the MZT, post-transcriptional gene regulation by RNA-binding proteins (RBPs) is the dominant force in embryo patterning. We used two mRNA interactome capture methods to identify RBPs bound to polyadenylated transcripts within the first 2 h of Drosophila melanogaster embryogenesis. We identified a high-confidence set of 476 putative RBPs and confirmed RNA-binding activities for most of 24 tested candidates. Most proteins in the interactome are known RBPs or harbor canonical RBP features, but 99 exhibited previously uncharacterized RNA-binding activity. mRNA-bound RBPs and TFs exhibit distinct expression dynamics, in which the newly identified RBPs dominate the first 2 h of embryonic development. Integrating our resource with in situ hybridization data from existing databases showed that mRNAs encoding RBPs are enriched in posterior regions of the early embryo, suggesting their general importance in posterior patterning and germ cell maturation.

In Vitro Spermatogenesis: How Far from Clinical Application?

Male infertility affects 7% of the male population, and 10% of infertile men are azoospermic. In these instances, using microsurgical testicular sperm extraction (m-TESE) and intra-cytoplasmic sperm injection (ICSI) helps a significant number of patients. However, in vitro differentiation of diploid germ cells to mature haploid germ cell has the potential to benefit many others, including pediatric cancer survivors who have previously cryopreserved their immature testicular tissue prior to starting gonadotoxic cancer treatment as well as men with spermatogenic arrest. This systematic review evaluates and summarizes half a century of researchers' efforts towards achieving in vitro spermatogenesis in mammalian species. A myriad of experimental assays and approaches has been developed using whole testis tissue or separated single cells from testis in two- or three-dimensional cell culture systems (2D versus 3D). Recent advances in the mammalian in vitro spermatogenesis, particularly in murine and nonhuman primate systems, hold promise towards translating the availability of in vitro spermatogenesis models in the human clinical setting in the near future.

MP70-05 DEVELOPMENT OF A NOVEL 3 DIMENSIONAL (3D) TESTICULAR ORGANOID MODEL FOR THE STUDY OF HUMAN SPERMATOGENESIS AND GENOTOXICITY IN VITRO

You have accessJournal of UrologyInfertility: Basic Research & Pathophysiology1 Apr 2016MP70-05 DEVELOPMENT OF A NOVEL 3 DIMENSIONAL (3D) TESTICULAR ORGANOID MODEL FOR THE STUDY OF HUMAN SPERMATOGENESIS AND GENOTOXICITY IN VITRO Samuel Pendergraft, Hooman Sadri-Ardekani, Tanya Reid, Anthony Atala, and Colin Bishop Samuel PendergraftSamuel Pendergraft More articles by this author , Hooman Sadri-ArdekaniHooman Sadri-Ardekani More articles by this author , Tanya ReidTanya Reid More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and Colin BishopColin Bishop More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1430AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Mammalian spermatogenesis is regulated through paracrine-endocrine activity, cell signaling, and local control mechanisms. These highly specific signaling interactions are effectively absent upon placing testicular cells into two-dimensional (2D) primary culture. Current protocols to produce mature germ cells in vitro are inefficient and are limited in supporting post-meiotic cells. In order to address these limitations we have developed a 3D testis organoid in vitro by combining stem cell and tissue engineering approaches. This model can be utilized as a means to evaluate gonadotoxic agents, and to address critical deficiencies in our understanding of basic human spermatogenesis Human testes tissues for this study were obtained through the National Disease Research Interchange (NDRI). METHODS Development of this model system consisted of (1) Identification and analysis of specific cellular components necessary for use in a 3D culture method, (2) establishment of basic design parameters, culture conditions, and characterization of human testicular organoids using live cell imaging, immunofluorescence, immunohistochemistry, gene expression, and viability assays, and (3) assess the potential to demonstrate testicular organoid function and as a tool for screening for testicular drug toxicity. RESULTS Human Spermatogonial stem cells (SSCs), Sertoli, and Leydig cells were isolated from human adult testes tissues, characterized, and expanded. These cell types were integrated successfully into 3D organoids and maintained viability as determined by ATP and Live/Dead assays for over 3 weeks in culture. These organoids were responsive to hCG and able to produce testosterone in vitro. Changes in gene expression within these multicellular human testis organoids was measured over time for cell and stage-specific markers including UCHL1, ITGA6, DAZL, SYCP3, PRM1, ACROSIN, FSHr, CYP19a1, HSD3B1, and CYP11a1. Dose response curves were generated using a luminescent viability assay in response to exposure to 3 commonly used genotoxic chemotherapeutic drugs and dose-response was compared to corresponding cells cultured in 2D. Calculated IC50 values were determined to be significantly higher for 3D organoid cultures. CONCLUSIONS Functional human 3D testicular organoids were successfully generated. Future directions include optimizing the spermatogenic capacity of the organoids and continuing to evaluate their use as a novel testicular toxicity model © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e907 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Samuel Pendergraft More articles by this author Hooman Sadri-Ardekani More articles by this author Tanya Reid More articles by this author Anthony Atala More articles by this author Colin Bishop More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Autophagy is required for ectoplasmic specialization assembly in sertoli cells

ABSTRACTThe ectoplasmic specialization (ES) is essential for Sertoli-germ cell communication to support all phases of germ cell development and maturity. Its formation and remodeling requires rapid reorganization of the cytoskeleton. However, the molecular mechanism underlying the regulation of ES assembly is still largely unknown. Here, we show that Sertoli cell-specific disruption of autophagy influenced male mouse fertility due to the resulting disorganized seminiferous tubules and spermatozoa with malformed heads. In autophagy-deficient mouse testes, cytoskeleton structures were disordered and ES assembly was disrupted. The disorganization of the cytoskeleton structures might be caused by the accumulation of a negative cytoskeleton organization regulator, PDLIM1, and these defects could be partially rescued by Pdlim1 knockdown in autophagy-deficient Sertoli cells. Altogether, our works reveal that the degradation of PDLIM1 by autophagy in Sertoli cells is important for the proper assembly of the ES, and these findings define a novel role for autophagy in Sertoli cell-germ cell communication.

Is there a role for DAZL in human female fertility?

The RNA binding protein deleted in azoospermia-like (Dazl) is a key determinant of germ cell maturation and entry into meiosis in rodents and other animal species. Although the complex phenotype of Dazl deficiency in both sexes, with defects at multiple stages of germ cell development and during meiosis, demonstrates its obligate significance in fertility in animal models, its involvement in human fertility is less clear. As an RNA binding protein, identification of the in vivo mRNA targets of DAZL is necessary to understand its influence. Thus far, only a small number of Dazl targets have been identified, which typically have pivotal roles in germ cell development and meiotic progression. However, it is likely that there are a number of additional germ cell and meiosis-relevant transcripts whose translation is affected in the absence of Dazl. Efforts to identify these RNA targets have mainly been focused on spermatogenesis, and restricted to mouse. In women, prophase I occurs in fetal life and it is during this period that the ovarian follicle pool is established, thus factors that have a role in determining the quality and quantity of the ovarian reserve may have significant impact on reproductive outcomes later in adult life. Here, we suggest that DAZL may be one such factor, and there is a need for greater understanding of the role of DAZL in human oogenesis and its contribution to lifelong female fertility.

Increased frequency of chromosome congression defects and aneuploidy in mouse oocytes cultured at lower temperature.

Optimal culture conditions are essential for successful IVM of mammalian oocytes and for their further development into an embryo. In the present study we used live cell imaging microscopy to assess the effects of suboptimal culture temperature on various aspects of IVM, including duration of meiosis I, dynamics of polar body extrusion, chromosome congression, anaphase-promoting complex/cyclosome (APC/C) activation and aneuploidy. The data showed that even a small deviation from the optimal incubation temperature causes marked changes in the duration and synchronicity of meiosis, APC/C activity and the frequency of chromosome congression and segregation errors. In vitro manipulation and maturation of germ cells is widely used in both human and animal artificial reproduction techniques. Mammalian oocytes are naturally prone to chromosomal segregation errors, which are responsible for severe mental and developmental disorders. The data presented herein demonstrate that exposure of mouse oocytes to suboptimal temperature during manipulation and maturation could further increase the frequency of chromosome segregation defects in these cells.

Presence of gonadotropin-releasing hormone-like peptide in the central nervous system and reproductive organs of the male blue swimming crab, Portunus pelagicus, and its effect on spermatogenesis.

Our previous studies have demonstrated that lamprey gonadotropin-releasing hormone-III (lGnRH-III)-like peptide occurs in the central nervous system (CNS) of decapod crustaceans (Macrobrachium rosenbergii, Penaeus monodon, Portunus pelagicus), and that lGnRH-III is the most potent in stimulating ovarian maturation compared with other GnRH isoforms. In this study, we examined the localization of lGnRH-III-like peptide in the CNS and male reproductive organs of the blue swimming crab by using anti-lGnRH-III as a probe. In the brain, lGnRH-III immunoreactivity (-ir) was detected in neurons of clusters 6, 10, 11, 14/15, 16, and 17 and in many neuropils. In the subesophageal ganglion, lGnRH-III-ir was present in neurons of the dorso-lateral and ventro-medial clusters. In the thoracic ganglia, lGnRH-III-ir was observed in the large-sized neurons between the thoracic neuropils and in the ventromedial cluster of the abdominal ganglia. In the testis, lGnRH-III-ir was detected in nurse cells, hemocytes, spermatids 2, and the outer and inner zones of the acrosomes of spermatozoa. Bioassay showed that lGnRH-III significantly increased the testis-somatic index, the percentage of late stages of seminiferous tubules (stages VII-IX), the diameter of the seminiferous tubules, and the number of BrdU-labeled early germ cells compared with the control groups. Thus, lGnRH-III-like peptide exists in the male crab and possibly enhances germ cell proliferation and maturation in the testes, leading to increased sperm production.

Cyclin-dependent kinase 4/6 inhibition for the treatment of mature teratoma germ cell tumor: Long-term follow-up of a phase II study.

474 Background: We have previously reported the results of a phase II clinical trial evaluating the safety and efficacy of palbociclib for the treatment of patients with unresectable germ cell tumors, including mature teratoma, teratoma with malignant transformation, and non-teratomatous tumors. In an effort to better assess the efficacy of palbociclib, we now report long-term follow-up of a more homogeneous population of patients with unresectable mature teratoma treated on the phase II clinical trial, with an emphasis on clinically-relevant endpoints for this incurable patient population. Methods: This was a retrospective analysis with long-term follow-up of the patient cohort with mature teratoma treated on the phase II study of palbociclib for the treatment of retinoblastoma protein-expressing germ cell tumors. Patient clinical data was obtained from the medical records and from communication with the enrolled patients and referring medical providers. Major medical events for the treatment of germ cell tumor, including before, during, and after study treatment, were recorded. Clinical events of interest included the initiation of systemic therapy, radiation therapy, surgical debulking, or other invasive procedures. Study endpoints included pre-study and post-study clinical event rates, event-free survival, and radiographic progression-free survival. Results: Long-term follow-up was obtained for 12 patients with mature teratoma. The median pre-study follow-up time was 19.7 months, and the median post-study follow-up time was 38.0 months. The median number of palbociclib treatment cycles was 11. The pre-study clinical event rate was 2.27 events/year (95% CI 1.66 – 3.13), and the post-study event rate was 0.62 events/year (95% CI 0.36 – 1.09). The median progression-free survival was 5.3 months (95% CI 1.8 – 22.6), and the median event-free survival was 16.2 months (95% CI 3.0 – 24.3). Conclusions: The initiation of palbociclib resulted in a clinically-meaningful delay in disease-related major clinical events and radiographic progression. These findings lend further support to the therapeutic activity of CDK4/6 inhibition in this incurable patient population.

Biology of the Sertoli Cell in the Fetal, Pubertal, and Adult Mammalian Testis.

A healthy man typically produces between 50 × 10(6) and 200 × 10(6) spermatozoa per day by spermatogenesis; in the absence of Sertoli cells in the male gonad, this individual would be infertile. In the adult testis, Sertoli cells are sustentacular cells that support germ cell development by secreting proteins and other important biomolecules that are essential for germ cell survival and maturation, establishing the blood-testis barrier, and facilitating spermatozoa detachment at spermiation. In the fetal testis, on the other hand, pre-Sertoli cells form the testis cords, the future seminiferous tubules. However, the role of pre-Sertoli cells in this process is much less clear than the function of Sertoli cells in the adult testis. Within this framework, we provide an overview of the biology of the fetal, pubertal, and adult Sertoli cell, highlighting relevant cell biology studies that have expanded our understanding of mammalian spermatogenesis.

Causes of hypogonadotropic hypogonadism predict response to gonadotropin substitution in adults.

Germ cell and Sertoli cell proliferation and maturation in human testes occur in three main waves, during the late fetal and early neonatal period and at early puberty. They are triggered by periods of increased activity of the hypothalamic-pituitary-gonadal (HPG) axis. In hypogonadotropic hypogonadism (HH), these processes are variably disturbed. The objective of this study was to explore whether success of gonadotropin replacement in HH men is predictable by the origin of HH, indicating time of onset and severity of GnRH/gonadotropin deficiency. The data of 51 adult HH patients who had undergone one cycle of hCG/FSH treatment were reviewed. Five groups were established, according to the underlying HH origin. Therapeutic success by final bi-testicular volumes (BTVs) final sperm concentrations (SC) and conception rates were compared and related to baseline parameters, indicative of the degree of HPG-axis disruption. Overall, BTVs rose from 13 ± 15 to 27 ± 15 mL, spermatogenesis was induced in 98%, with mean SCs of 15 ± 30 mill/mL, spontaneous pregnancies in 37% and additional 18% via intracytoplasmic sperm injection. Kallmann syndrome patients had the poorest responses (BTV: 16.9 ± 10 mL; SC: 3.5 ± 5.6 mill/mL), followed by patients with congenital/infancy-acquired multiple pituitary hormone deficiencies (MPHD) and patients with HH+absent puberty (BTV: 21 ± 14/24 ± 9 mL; SC: 5.5 ± 6.5/ 14.5 ± 23.8 mill/mL). HH men with pubertal arrest and with post-pubertally acquired MPHD had the best results (BTV: 36 ± 14/38 ± 16 mL; SC: 25.4 ± 34.2/29.9 ± 50.5 mill/mL). Earlier conception after 20.3 ± 11.5 months (vs. 43.1 ± 43.8; p = 0.047) of gonadotropin treatment with higher pregnancy rates (62% vs. 42%) was achieved in the two post-pubertally acquired HH subgroups, compared to the three pre-pubertally acquired. Therapeutic success was higher in patients without previously undescended testes, with higher baseline BTVs (pre- vs. post-pubertal HH: 5 ± 4 mL vs. 26 ± 16 mL; p < 0.0001) and higher baseline inhibinB levels (pre- vs. post-pubertal HH: 16.6 vs. 144.5 pg/mL; p = 0.0004). The cause of HH is a valuable predictor of outcome of gonadotropin replacement in adults.

Annual Reproductive Cycle and Unusual Embryogenesis of a Temperate Coral in the Mediterranean Sea.

The variety of reproductive processes and modes among coral species reflects their extraordinary regeneration ability. Scleractinians are an established example of clonal animals that can exhibit a mixed strategy of sexual and asexual reproduction to maintain their populations. This study provides the first description of the annual reproductive cycle and embryogenesis of the temperate species Caryophyllia inornata. Cytometric analyses were used to define the annual development of germ cells and embryogenesis. The species was gonochoric with three times more male polyps than female. Polyps were sexually mature from 6 to 8 mm length. Not only females, but also sexually inactive individuals (without germ cells) and males were found to brood their embryos. Spermaries required 12 months to reach maturity, while oogenesis seemed to occur more rapidly (5–6 months). Female polyps were found only during spring and summer. Furthermore, the rate of gamete development in both females and males increased significantly from March to May and fertilization was estimated to occur from April to July, when mature germ cells disappeared. Gametogenesis showed a strong seasonal influence, while embryos were found throughout the year in males and in sexually inactive individuals without a defined trend. This unusual embryogenesis suggests the possibility of agamic reproduction, which combined with sexual reproduction results in high fertility. This mechanism is uncommon and only four other scleractinians (Pocillopora damicornis, Tubastraea diaphana, T. coccinea and Oulastrea crispata) have been shown to generate their broods asexually. The precise nature of this process is still unknown.

Molecular, cellular, morphological, physiological and behavioral aspects of gonadotropin-inhibitory hormone

Gonadotropin-inhibitory hormone (GnIH) is a hypothalamic neuropeptide that was isolated from the brains of Japanese quail in 2000, which inhibited luteinizing hormone release from the anterior pituitary gland. Here, we summarize the following fifteen years of researches that investigated on the mechanism of GnIH actions at molecular, cellular, morphological, physiological, and behavioral levels. The unique molecular structure of GnIH peptide is in its LPXRFamide (X=L or Q) motif at its C-terminal. The primary receptor for GnIH is GPR147. The cell signaling pathway triggered by GnIH is initiated by inhibiting adenylate cyclase and decreasing cAMP production in the target cell. GnIH neurons regulate not only gonadotropin synthesis and release in the pituitary, but also regulate various neurons in the brain, such as GnRH1, GnRH2, dopamine, POMC, NPY, orexin, MCH, CRH, oxytocin, and kisspeptin neurons. GnIH and GPR147 are also expressed in gonads and they may regulate steroidogenesis and germ cell maturation in an autocrine/paracrine manner. GnIH regulates reproductive development and activity. In female mammals, GnIH may regulate estrous or menstrual cycle. GnIH is also involved in the regulation of seasonal reproduction, but GnIH may finely tune reproductive activities in the breeding seasons. It is involved in stress responses not only in the brain but also in gonads. GnIH may inhibit male socio-sexual behavior by stimulating the activity of cytochrome P450 aromatase in the brain and stimulates feeding behavior by modulating the activities of hypothalamic and central amygdala neurons.