Lack Germ Cells Research Articles

The histone demethylase KDM1A is essential for the maintenance and differentiation of spermatogonial stem cells and progenitors.

Little is known of the fundamental processes governed by epigenetic mechanisms in the supplier cells of spermatogenesis, the spermatogonial stem cells (SSCs). The histone H3 lysine demethylase KDM1A is expressed in spermatogonia. We hypothesized that KDM1A serves in transcriptional regulation of SSCs and fertility. Using a conditional deletion of Kdm1a [conditional knockout (cKO)] in mouse spermatogonia, we determined that Kdm1a is essential for spermatogenesis as adult cKO males completely lack germ cells. Analysis of postnatal testis development revealed that undifferentiated and differentiating spermatogonial populations form in Kdm1a-cKO animals, yet the majority fail to enter meiosis. Loss of germ cells in the cKO was rapid with none remaining by postnatal day (PND) 21. To gain insight into the mechanistic implications of Kdm1a ablation, we isolated PND 6 spermatogonia enriched for SSCs and analyzed their transcriptome by RNA sequencing. Loss of Kdm1a was associated with altered transcription of 1206 genes. Importantly, differentially expressed genes between control and Kdm1a-cKO animals included those that are essential for SSC and progenitor maintenance and spermatogonial differentiation. The complete loss of fertility and failure to establish spermatogenesis indicate that Kdm1a is a master controller of gene transcription in spermatogonia and is required for SSC and progenitor maintenance and differentiation.

TAF4b is required for mouse spermatogonial stem cell development.

Long-term mammalian spermatogenesis requires proper development of spermatogonial stem cells (SSCs) that replenish the testis with germ cell progenitors during adult life. TAF4b is a gonadal-enriched component of the general transcription factor complex, TFIID, which is required for the maintenance of spermatogenesis in the mouse. Successful germ cell transplantation assays into adult TAF4b-deficient host testes suggested that TAF4b performs an essential germ cell autonomous function in SSC establishment and/or maintenance. To elucidate the SSC function of TAF4b, we characterized the initial gonocyte pool and rounds of spermatogenic differentiation in the context of the Taf4b-deficient mouse testis. Here, we demonstrate a significant reduction in the late embryonic gonocyte pool and a deficient expansion of this pool soon after birth. Resulting from this reduction of germ cell progenitors is a developmental delay in meiosis initiation, as compared to age-matched controls. While GFRα1+ spermatogonia are appropriately present as Asingle and Apaired in wild-type testes, TAF4b-deficient testes display an increased proportion of long and clustered chains of GFRα1+ cells. In the absence of TAF4b, seminiferous tubules in the adult testis either lack germ cells altogether or are found to have missing generations of spermatogenic progenitor cells. Together these data indicate that TAF4b-deficient spermatogenic progenitor cells display a tendency for differentiation at the expense of self-renewal and a renewing pool of SSCs fail to establish during the critical window of SSC development.

Emerging methods to generate artificial germ cells from stem cells.

Germ cells are responsible for the transmission of genetic and epigenetic information across generations. At present, the number of infertile couples is increasing worldwide; these infertility problems can be traced to environmental pollutions, infectious diseases, cancer, psychological or work-related stress, and other factors, such as lifestyle and genetics. Notably, lack of germ cells and germ cell loss present real obstacles in infertility treatment. Recent research aimed at producing gametes through artificial germ cell generation from stem cells may offer great hope for affected couples to treat infertility in the future. Therefore, this rapidly emerging area of artificial germ cell generation from nongermline cells has gained considerable attention from basic and clinical research in the fields of stem cell biology, developmental biology, and reproductive biology. Here, we review the state of the art in artificial germ cell generation.

A germ cell determinant reveals parallel pathways for germ line development in Caenorhabditis elegans.

Despite the central importance of germ cells for transmission of genetic material, our understanding of the molecular programs that control primordial germ cell (PGC) specification and differentiation are limited. Here, we present findings that X chromosome NonDisjunction factor-1 (XND-1), known for its role in regulating meiotic crossover formation, is an early determinant of germ cell fates in Caenorhabditis elegans. xnd-1 mutant embryos display a novel 'one PGC' phenotype as a result of G2 cell cycle arrest of the P4 blastomere. Larvae and adults display smaller germ lines and reduced brood size consistent with a role for XND-1 in germ cell proliferation. Maternal XND-1 proteins are found in the P4 lineage and are exclusively localized to the nucleus in PGCs, Z2 and Z3. Zygotic XND-1 turns on shortly thereafter, at the ∼300-cell stage, making XND-1 the earliest zygotically expressed gene in worm PGCs. Strikingly, a subset of xnd-1 mutants lack germ cells, a phenotype shared with nos-2, a member of the conserved Nanos family of germline determinants. We generated a nos-2 null allele and show that nos-2; xnd-1 double mutants display synthetic sterility. Further removal of nos-1 leads to almost complete sterility, with the vast majority of animals without germ cells. Sterility in xnd-1 mutants is correlated with an increase in transcriptional activation-associated histone modification and aberrant expression of somatic transgenes. Together, these data strongly suggest that xnd-1 defines a new branch for PGC development that functions redundantly with nos-2 and nos-1 to promote germline fates by maintaining transcriptional quiescence and regulating germ cell proliferation.

Molecular Reproduction & Development Volume 81, Issue 10, October 2014 cover image

In the absence of germ cells, the zebrafish gonad develops as an empty somatic testis. Hartung et. al. (this issue) demonstrate that Vasa is essential for progression beyond the pachytene stage of meiosis by a tumor protein 53 (tp53)‐independent mechanism. This image shows a hematoxylin‐and‐eosin‐stained, transverse section through the trunk of day‐42 vasa;tp53 double‐mutant fish. The somatic cells of the gonads, which lack germ cells at this stage, are seen above the intestine.

In utero exposure to the oestrogen mimic diethylstilbestrol disrupts gonadal development in a viviparous reptile

The ubiquitous presence of endocrine-disrupting chemicals (EDCs) in the environment is of major concern. Studies on oviparous reptiles have significantly advanced knowledge in this field; however, 30% of reptilian species are viviparous (live-bearing), a parity mode in which both yolk and a placenta support embryonic development, thus exposure to EDCs may occur via multiple routes. In this first study of endocrine disruption in a viviparous lizard (Niveoscincus metallicus), we aimed to identify effects of the oestrogen mimic diethylstilbestrol (DES) on gonadal development. At the initiation of sexual differentiation, pregnant N. metallicus were treated with a single dose of DES at 100 or 10µgkg(--1), a vehicle solvent or received no treatment. There was no dose-response effect, but the testes of male neonates born to DES-exposed mothers showed reduced organisation of seminiferous tubules and a lack of germ cells compared with those from control groups. The ovaries of female neonates born to DES-exposed mothers exhibited phenotypic abnormalities of ovarian structure, oocytes and follicles compared with controls. The results indicate that, in viviparous lizards, maternal exposure to oestrogenic EDCs during gestation may have profound consequences for offspring reproductive fitness.

Predictive factors of successful sperm retrieval in azoospermia

IntroductionTesticular sperm extraction with intracytoplasmic sperm injection is the standard treatment for azoospermia. The objective of this study is to identify predictive factors of successful sperm retrieval. Materials and methodsBetween June 2003 and May 2011, we tried testicular sperm extraction (TESE) in 74 azoospermic patients in the Reproductive Medicine Unit of Son Espases Hospital (Palma de Mallorca). Serum follicle stimulating hormone (FSH) and inhibin B levels, testicular histology, genetic study, presence or not of cryptozoospermia and testicular volume were examined. ResultsSpermatozoa were successfully recovered in 47.2% of the total patients, in 36% of non-obstructive azoospermic patients and in 100% of obstructive azoospermic patients. Low inhibin B and high FSH were correlated to sperm retrieval failure. The cutoff points were determined using ROC curves that were 67pg/mL for inhibin B and 12.2mUI/mL for FSH. Spermatozoa were not successfully retrieved in any patient with Y microdeletions in AZFa,b regions. Spermatozoa were successfully retrieved in 100% of the patients with CFTR mutations. The highest sperm retrieval rate was for hypospermatogenesis, followed by maturation arrest and Sertoli-cell-only. Spermatozoa were successfully retrieved in all cryptozoospermic patients. Although using a non-significant test, there seems to be a correlation between higher testicular volume and a higher probability of successful sperm retrieval. ConclusionsExcept for Y microdeletions in AZFa,b regions, there is no predictive factor of testicular sperm retrieval to rule out a patient for TESE. Lower inhibin B is more related to sperm retrieval failure than higher FSH. Sperm retrieval is possible for all cases of CFTR mutations but in any case of microdeletion Y in AZFa,b. The lack of germ cells is correlated with a high probability of sperm retrieval failure. The presence of cryptozoospermia is correlated with a high probability of sperm retrieval success. We do not find a statistically significant relation between testicular volume and successful sperm retrieval.

Germ cells are required to maintain a stable sexual phenotype in adult zebrafish

Sex in zebrafish is not determined by a major chromosomal locus, but instead relies on a mechanism that is influenced by a germ cell-derived signal, as animals that lack germ cells, or specifically oocytes, develop as phenotypic males. These data suggest that during primary sex determination, an oocyte-derived signal acts on the bipotential somatic gonad to promote the female-specific program. However, it is not known if germ cells are required only during the primary sex-determining window, or if they are required throughout adult life to maintain the female sexual phenotype. Here, we show that while wild-type zebrafish do not switch sex as adults, germ cell-depleted adult females readily convert to a male phenotype. Notably, when oocytes are depleted, but germline stem cells remain, adult females sex-revert to sperm-producing males, indicating that a germ cell-derived signal acts on the somatic gonad to promote female development directly or indirectly by repressing male-specific gene expression. These results also confirm that signals from the somatic gonad in turn ensure that the sex appropriate gamete is produced.

Mutations of NANOS1, a human homologue of the Drosophila morphogen, are associated with a lack of germ cells in testes or severe oligo-astheno-teratozoospermia

BackgroundThe Nanos gene is a key translational regulator of specific mRNAs involved in Drosophila germ cell development. Disruption of mammalian homologues, Nanos2 or Nanos3, causes male infertility in mice. In...

Factores predictivos de recuperación espermática en las azoospermias

IntroductionTesticular sperm extraction with intracytoplasmic sperm injection is the standard treatment for azoospermia. The objective of this study is to identify predictive factors of successful sperm retrieval. Materials and methodsBetween June 2003 and May 2011, we tried testicular sperm extraction (TESE) in 74 azoospermic patients in the Reproductive Medicine Unit of Son Espases Hospital (Palma de Mallorca). Serum follicle stimulating hormone (FSH) and inhibin B levels, testicular histology, genetic study, presence or not of cryptozoospermia and testicular volume were examined. ResultsSpermatozoa were successfully recovered in 47.2% of the total patients, in 36% of non-obstructive azoospermic patients and in 100% of obstructive azoospermic patients. Low inhibin B and high FSH were correlated to sperm retrieval failure. The cutoff points were determined using ROC curves that were 67 pg/mL for inhibin B and 12.2 mUI/mL for FSH. Spermatozoa were not successfully retrieved in any patient with Y microdeletions in AZFa,b regions. Spermatozoa were successfully retrieved in 100% of the patients with CFTR mutations. The highest sperm retrieval rate was for hypospermatogenesis, followed by maturation arrest and Sertoli-cell-only. Spermatozoa were successfully retrieved in all cryptozoospermic patients. Although using a non-significant test, there seems to be a correlation between higher testicular volume and a higher probability of successful sperm retrieval. ConclusionsExcept for Y microdeletions in AZFa,b regions, there is no predictive factor of testicular sperm retrieval to rule out a patient for TESE. Lower inhibin B is more related to sperm retrieval failure than higher FSH. Sperm retrieval is possible for all cases of CFTR mutations but in any case of microdeletion Y in AZFa,b. The lack of germ cells is correlated with a high probability of sperm retrieval failure. The presence of cryptozoospermia is correlated with a high probability of sperm retrieval success. We do not find a statistically significant relation between testicular volume and successful sperm retrieval.

Germ cells are not the primary factor for sexual fate determination in goldfish

The presence of germ cells in the early gonad is important for sexual fate determination and gonadal development in vertebrates. Recent studies in zebrafish and medaka have shown that a lack of germ cells in the early gonad induces sex reversal in favor of a male phenotype. However, it is uncertain whether the gonadal somatic cells or the germ cells are predominant in determining gonadal fate in other vertebrate. Here, we investigated the role of germ cells in gonadal differentiation in goldfish, a gonochoristic species that possesses an XX–XY genetic sex determination system. The primordial germ cells (PGCs) of the fish were eliminated during embryogenesis by injection of a morpholino oligonucleotide against the dead end gene. Fish without germ cells showed two types of gonadal morphology: one with an ovarian cavity; the other with seminiferous tubules. Next, we tested whether function could be restored to these empty gonads by transplantation of a single PGC into each embryo, and also determined the gonadal sex of the resulting germline chimeras. Transplantation of a single GFP-labeled PGC successfully produced a germline chimera in 42.7% of the embryos. Some of the adult germline chimeras had a developed gonad on one side that contained donor derived germ cells, while the contralateral gonad lacked any early germ cell stages. Female germline chimeras possessed a normal ovary and a germ-cell free ovary-like structure on the contralateral side; this structure was similar to those seen in female morphants. Male germline chimeras possessed a testis and a contralateral empty testis that contained some sperm in the tubular lumens. Analysis of aromatase, foxl2 and amh expression in gonads of morphants and germline chimeras suggested that somatic transdifferentiation did not occur. The offspring of fertile germline chimeras all had the donor-derived phenotype, indicating that germline replacement had occurred and that the transplanted PGC had rescued both female and male gonadal function. These findings suggest that the absence of germ cells did not affect the pathway for ovary or testis development and that phenotypic sex in goldfish is determined by somatic cells under genetic sex control rather than an interaction between the germ cells and somatic cells.

The C. elegans MicroRNA mir-71 Acts in Neurons to Promote Germline-Mediated Longevity through Regulation of DAF-16/FOXO

The life span of Caenorhabditis elegans is controlled by signaling between the germline and the soma. Germ cell removal extends life span by triggering the activation of the DAF-16/FOXO transcription factor in the intestine. Here we analyze microRNA function in C.elegans aging and show that the microRNA mir-71 functions to mediate the effects of germ cell loss on life span. mir-71 is required for the life span extension caused by germline removal, and overexpression of mir-71 further extends the life span of animals lacking germ cells. mir-71 functions in the nervous system to facilitate the localization and transcriptional activity of DAF-16 in the intestine. Our findings reveal a microRNA-dependent mechanism of life span regulation by the germline and indicate that signaling among the gonad, the nervous system, and the intestine coordinates the life span of the entire organism.

Rescue of tropomyosin deficiency inDrosophilaand human cancer cells by synaptopodin reveals a role of tropomyosin α in RhoA stabilization

Tropomyosins are widespread actin-binding proteins that influence numerous cellular functions including actin dynamics, cell migration, tumour suppression, and Drosophila oocyte development. Synaptopodin is another actin-binding protein with a more restricted expression pattern in highly dynamic cell compartments such as kidney podocyte foot processes, where it promotes RhoA signalling by blocking the Smurf1-mediated ubiquitination of RhoA. Here, we show that synaptopodin has a shorter half-life but shares functional properties with the highly stable tropomyosin. Transgenic expression of synaptopodin restores oskar mRNA localization in Drosophila oocytes mutant for TmII, thereby rescuing germline differentiation and fertility. Synaptopodin restores stress fibres in tropomyosin-deficient human MDA-MB 231 breast cancer cells and TPMα-depleted fibroblasts. Gene silencing of TPMα but not TPMβ causes loss of stress fibres by promoting Smurf1-mediated ubiquitination and proteasomal degradation of RhoA. Functionally, overexpression of synaptopodin or RhoA(K6,7R) significantly reduces MDA-MB 231 cell migration. Our findings elucidate RhoA stabilization by structurally unrelated actin-binding proteins as a conserved mechanism for regulation of stress fibre dynamics and cell motility in a cell type-specific fashion.

Reconstituting mammalian spermatogenesis using stem cells

While stock markets and economies are smoldering all over the world, stem cell science is on fire. The promise of curing many untreatable forms of human male infertility is now one small step closer based on a recent paper in Cell.1 In this work, scientists from Kyoto University in Japan created mature, fertile sperm from embryonic and induced pluripotent stem cells by grabbing the primitive germ cells called primordial germ cells just as they were being made from their pluripotent precursors in vitro and transplanting them back into genetically sterile mice. As background, stem cells possess the unique ability to either propagate by self-renewal or to differentiate to mature tissues under the influence of appropriate molecular cues. This remarkable dual behavior has focused the research spotlight on using stem cells for regenerative medicine. There are now several classes of stem cells, all varying in degrees of pluripotency.2 Among these, embryonic stem cells represent the prototypical pluripotent stem cells. Another, more recent stem cell is derived from the reprogramming of somatic cells to pluripotent stem cells (iPSCs) and is based on a set of landmark studies conducted by Yamanaka and others3 that showed that just four factors (Oct4, Sox2, Klf4 and c-Myc) normally produced in the embryo are sufficient to reprogram differentiated, non-embryonic cells into pluripotent cells. In addition, during development, a small population of embryonic stem cells are allocated to the germ line (primordial germ cells), while the remaining cells give rise to the three germ layers. It is this particular stem cell type that has now captured the research limelight. Over the past 2 years, germ line stem cell research has seen spectacular advance which suggests that these particular stem cells are quite plastic and manipulatable for potential cell-based therapy. Some of these advances include: The demonstration by several groups that both adult mouse and human testis stem cells, termed spermatogonial stem cells, can be ‘induced' or ‘transformed' into embryonic-like stem cells.4, 5, 6 The observation that both human embryonic and adult-induced pluripotency stem cells can be directed down the pathway of germ cell differentiation through meiosis in vitro.7, 8 The finding that neonatal, mouse, testis stem cells (gonocytes) can placed in an in vitro organ culture system and be prodded into making mature, fertile sperm.9 The report that both pre- and post-pubertal human spermatogonial stem cells can be propagated in vitro for prolonged periods of time.10, 11 In the Kyoto study, both embryonic stem cells and iPSCs from mice were observed as they began to differentiate into other cell types in vitro. From developing epiblast-like cells, using global transcriptome profiling, information from epigenetic programming and cellular dynamics, they defined the most primitive of true germ cells, the differentiating primordial germ cells (PGCs). They observed that PGCs are indeed distinct from epiblast cells and also noted that they are transient in the germ cell lineage. When properly characterized and injected into the testes of genetically (W/Wv) mice, the PGCs formed competent sperm 50% of the time. The sperm was also capable of producing normal F1 mice after assisted reproduction. And, the F1 mice were naturally fertile! This success took a lot of work, and invoked a host of molecular and expression characterization of developing cells and lots of sweat equity. And not all the results were rosy. When cells slightly different from primordial germ cells were injected into sterile testicles, benign tumors formed, the scourge of stem cell science. This emphasizes the importance of identifying putative surface markers that delineate a pure population of PGCs for use with this approach. Not only that, when iPSCs were used to create PGCs and transplanted into sterile mice testes, some of the offspring died prematurely with ‘tumors around the neck region', a finding that will certainly will need further investigation if the technique is to become clinically viable in humans. Recreating mammalian spermatogenesis, an enormously complex process involving both mitosis and meiosis and multilayered quality control check points, is a scientific and clinical holy grail. Clinically, these techniques hold promise for creating man-made sperm for infertile men who lack germ cells for genetic or acquired (i.e. cancer treatment) reasons. If fully realized in humans, it may be possible to use non-germ cell sources to make sperm; maybe even a simple skin biopsy could be used as starting point. Saitou and colleagues have taken us one step closer to its realization by employing embryonic stem cells and iPSCs as precursors and functionally recapitulating the process in mice.

A late phase of germ plasm accumulation during Drosophila oogenesis requires Lost and Rumpelstiltskin

Asymmetric mRNA localization is an effective mechanism for establishing cellular and developmental polarity. Posterior localization of oskar in the Drosophila oocyte targets the synthesis of Oskar to the posterior, where Oskar initiates the assembly of the germ plasm. In addition to harboring germline determinants, the germ plasm is required for localization and translation of the abdominal determinant nanos. Consequently, failure of oskar localization during oogenesis results in embryos lacking germ cells and abdominal segments. oskar accumulates at the oocyte posterior during mid-oogenesis through a well-studied process involving kinesin-mediated transport. Through live imaging of oskar mRNA, we have uncovered a second, mechanistically distinct phase of oskar localization that occurs during late oogenesis and results in amplification of the germ plasm. Analysis of two newly identified oskar localization factors, Rumpelstiltskin and Lost, that are required specifically for this late phase of oskar localization shows that germ plasm amplification ensures robust abdomen and germ cell formation during embryogenesis. In addition, our results indicate the importance of mechanisms for adapting mRNAs to utilize multiple localization pathways as necessitated by the dramatic changes in ovarian physiology that occur during oogenesis.

Roles of brca2 (fancd1) in Oocyte Nuclear Architecture, Gametogenesis, Gonad Tumors, and Genome Stability in Zebrafish

Mild mutations in BRCA2 (FANCD1) cause Fanconi anemia (FA) when homozygous, while severe mutations cause common cancers including breast, ovarian, and prostate cancers when heterozygous. Here we report a zebrafish brca2 insertional mutant that shares phenotypes with human patients and identifies a novel brca2 function in oogenesis. Experiments showed that mutant embryos and mutant cells in culture experienced genome instability, as do cells in FA patients. In wild-type zebrafish, meiotic cells expressed brca2; and, unexpectedly, transcripts in oocytes localized asymmetrically to the animal pole. In juvenile brca2 mutants, oocytes failed to progress through meiosis, leading to female-to-male sex reversal. Adult mutants became sterile males due to the meiotic arrest of spermatocytes, which then died by apoptosis, followed by neoplastic proliferation of gonad somatic cells that was similar to neoplasia observed in ageing dead end (dnd)-knockdown males, which lack germ cells. The construction of animals doubly mutant for brca2 and the apoptotic gene tp53 (p53) rescued brca2-dependent sex reversal. Double mutants developed oocytes and became sterile females that produced only aberrant embryos and showed elevated risk for invasive ovarian tumors. Oocytes in double-mutant females showed normal localization of brca2 and pou5f1 transcripts to the animal pole and vasa transcripts to the vegetal pole, but had a polarized rather than symmetrical nucleus with the distribution of nucleoli and chromosomes to opposite nuclear poles; this result revealed a novel role for Brca2 in establishing or maintaining oocyte nuclear architecture. Mutating tp53 did not rescue the infertility phenotype in brca2 mutant males, suggesting that brca2 plays an essential role in zebrafish spermatogenesis. Overall, this work verified zebrafish as a model for the role of Brca2 in human disease and uncovered a novel function of Brca2 in vertebrate oocyte nuclear architecture.

Uncovering Gene Regulatory Networks During Mouse Fetal Germ Cell Development

The commitment of germ cells to either oogenesis or spermatogenesis occurs during fetal gonad development: germ cells enter meiosis or mitotic arrest, depending on whether they reside within an ovary or a testis, respectively. Despite the critical importance of this step for sexual reproduction, gene networks underlying germ cell development have remained only partially understood. Taking advantage of the W(v) mouse model, in which gonads lack germ cells, we conducted a microarray study to identify genes expressed in fetal germ cells. In addition to distinguishing genes expressed by germ cells from those expressed by somatic cells within the developing gonads, we were able to highlight specific groups of genes expressed only in female or male germ cells. Our results provide an important resource for deciphering the molecular pathways driving proper germ cell development and sex determination and will improve our understanding of the etiology of human germ cell tumors that arise from dysregulation of germ cell differentiation.

Abnormal gonad development in Kit W-2Bao mice caused by a Kit gene missense mutation

KitW-2Bao mice are single-gene autosomal dominant mutation mice with a B6 background that were bred in our laboratory. Heterozygotes had morphological characteristics including albinism of the abdomen, extremities, and tail, whereas the homozygotes had albinism of the body, black eyes, and infertility. The homozygous mutants showed small, structurally abnormal gonads, and lacked germ cells. Heterozygous male mice lacked germ cells in some contorted seminiferous tubules. This mutation has been mapped at 43.8 cM from the centromere in chromosome 5 by linkage analysis and Kit has been identified as the candidate gene. After Kit full-length mRNA amplification, it was found that a G to T conversion at position 1228 in the ORF changed the 410th amino acid from V to F. This amino acid change could affect the protein’s secondary structure. Heterozygous mutant mice were intercrossed and homozygous mutant mice were bred and genotyped. We found that no primordial germ cells (PGCs) appeared in the urogenital ridge area at fetus day 11.5 in the homozygotes. The number of PGCs also significantly decreased in heterozygotes. At fetus day 15.5, the differentiation of the testis tubule structure was unclear; as well, they contained no spermatogonia. Female homozygotes contained no primordial follicles in the ovary. The numbers of PGCs and primordial follicles were significantly decreased in heterozygous mice. W−2Bao is the only mutated site in the extracellular 4th Ig-like domain and this mutant mouse model provides new material for the study of the mechanism of reproductive system development.

Sexual dimorphism of gonadal structure and gene expression in germ cell-deficient loach, a teleost fish

Germ cell-deficient fish usually develop as phenotypic males. Thus, the presence of germ cells is generally considered to be essential for female gonadal differentiation or the maintenance of ovarian structure. However, little is known of the role of germ cells in the determination of the sexual fate of gonadal somatic cells. We have established an inducible germ cell deficiency system in the loach (Misgurnus anguillicaudatus, Cypriniformes: Cobitidae), a small freshwater fish, using knockdown of the dead end gene with a morpholino antisense oligonucleotide. Interestingly, loach lacking germ cells could develop as either phenotypic males or females, as characterized morphologically by the presence or absence of bony plates in the pectoral fins, respectively. The phenotypic males and females had testicular and ovarian structures, respectively, but lacked germ cells. Gene expression patterns in these male and female germ cell-deficient gonads were essentially the same as those in gonads of normal fish. Our observations indicate that sexually dimorphic gonads can develop in germ cell-deficient loach. In contrast to the situation in other model fish species, the gonadal somatic cells in phenotypic females autonomously differentiated into ovarian tissues and also played a role in the maintenance of gonadal structure. On the basis of our observations, we propose two possible models to explain the role of germ cells in sex determination in fish.

The Somatic Reproductive Tissues of C. elegans Promote Longevity through Steroid Hormone Signaling

Author SummaryReproductive tissues are known to generate important intercellular signals. For example, in mammals, the reproductive tissues produce steroid hormones such as estrogen and testosterone that have profound effects on development and physiology. Studies of the nematode C. elegans and other organisms have shown that the reproductive system can also affect the rate at which an animal ages. Removal of C. elegans' germ cells extends lifespan but this effect is not simply due to sterility, as removal of both the somatic reproductive tissues and the germ cells does not extend lifespan. Instead, loss of the germ cells extends lifespan by activating a pathway that requires input from the somatic gonad. In this study, we demonstrate that the somatic reproductive tissues promote longevity by controlling the activity of a steroid signaling pathway that regulates the DAF-12 nuclear hormone receptor.