Function of Nanos2 in the male germ cell lineage in mice

Nanos is known as an evolutionarily conserved RNA-binding protein, the function of which is implicated in germ cell development. This includes the maintenance of both the primordial germ cells (PGCs) and germline stem cells. In mice, Nanos2 exhibits a unique feature in which its expression is induced only in the germ cells within the sexually determined male gonad. Nanos2 promotes male germ cell differentiation, while simultaneously suppressing a female program. In addition, Nanos2 is also expressed in the spermatogonial stem cells and functions as an intrinsic factor to maintain the stem cell population during spermatogenesis. Detailed cytological and biochemical analyses in embryonic male gonads in the mouse have revealed that Nanos2 localizes to the P-bodies, a center of RNA processing. It has also been shown that the Nanos2 interacts with protein components of the deadenylation complex involved in the initial step of the RNA degradation pathway.     

Genetics of early mammalian folliculogenesis

Early ovarian folliculogenesis begins with the breakdown of germ cell clusters and formation of primordial follicles. Primordial follicles are the smallest ovarian follicle units continuously recruited to grow into primary and more advanced ovarian follicles. Genes expressed in the germ cells such as Figla, Nobox, Kit and Ntrk2, as well as genes expressed in the surrounding somatic cells such as Foxl2, Kitl and Ngf, play critical functions during early folliculogenesis. Transgenic mice continue to provide important insights into the genetic pathways that regulate early mammalian folliculogenesis. Genes critical in early folliculogenesis are important determinants of reproductive life span and represent candidate genes for human ovarian failure. Received 25 August 2005; received after revision 18 October 2005; accepted 21 November 2005     

Adenylate storage,metabolism and utilization in coelomic cells of the polychaeteNereis virens (Annelida,polychaeta)

Eleocytes are specialized coelomic cells in nereid annelids which assume a central role during germ cell development. They may contain extremely high concentrations of both adenosine monophosphate (AMP) and adenosine diphosphate (ADP) (each >10 mol/ml of cell vol.), whereas the adenosine triphosphate (ATP) content is comparatively low (0.8 mol/ml cell vol.).³¹P nuclear magnetic, resonance (NMR) studies of living eleocytes suggest the compartmentalization of both AMP and ADP in the large acidic vacuole characteristic for this cell type. Eleocytes are thus capable of storing high concentrations of ADP and AMP without inhibiting energy metabolism, by sequestering these compounds in a separate compartment. The high concentrations of both AMP and ADP in the eleocytes decrease in both males and females during the course of maturation. In eleocytes of male animals, the decline of the high nucleotide concentrations was accompanied by a transient increase of two intracellular nucleosides, inosine and guanosine. This suggests the degradation and further metabolism of nucleotides to the corresponding nucleosides. In culture, eleocytes release both inosine and guanosine into the medium. Both nucleosides are also present in the coelomic fluid, the common compartment for both eleocytes and germ cells. Both male and female germ cells incorporate¹⁴C-labelled inosine and guanosine in culture. For oocytes, the further incorporation of [¹⁴C]inosine into the RNA fraction could be demonstrated. The large adenylate pools in the eleocytes may be regarded as a store for purine compounds for later use by the growing germ cells to supplement nucleic acid synthesis. The supply of nucleic acid precursors seems to be another specific function of eleocytes related to gametogenesis, in addition to their known synthesis of vitellogenin.     

Regulation of male sex determination: genital ridge formation and Sry activation in mice

Sex determination is essential for the sexual reproduction to generate the next generation by the formation of functional male or female gametes. In mammals, primary sex determination is commenced by the presence or absence of the Y chromosome, which controls the fate of the gonadal primordium. The somatic precursor of gonads, the genital ridge is formed at the mid-gestation stage and gives rise to one of two organs, a testis or an ovary. The fate of the genital ridge, which is governed by the differentiation of somatic cells into Sertoli cells in the testes or granulosa cells in the ovaries, further determines the sex of an individual and their germ cells. Mutation studies in human patients with disorders of sex development and mouse models have revealed factors that are involved in mammalian sex determination. In most of mammals, a single genetic trigger, the Y-linked gene Sry (sex determination region on Y chromosome), regulates testicular differentiation. Despite identification of Sry in 1990, precise mechanisms underlying the sex determination of bipotential genital ridges are still largely unknown. Here, we review the recent progress that has provided new insights into the mechanisms underlying genital ridge formation as well as the regulation of Sry expression and its functions in male sex determination of mice.     

Hormonal control of Sertoli cell metabolism regulates spermatogenesis

Hormonal regulation is essential to spermatogenesis. Sertoli cells (SCs) have functions that reach far beyond the physical support of germ cells, as they are responsible for creating the adequate ionic and metabolic environment for germ cell development. Thus, much attention has been given to the metabolic functioning of SCs. During spermatogenesis, germ cells are provided with suitable metabolic substrates, in a set of events mediated by SCs. Multiple signaling cascades regulate SC function and several of these signaling pathways are hormone-dependent and cell-specific. Within the seminiferous tubules, only SCs possess receptors for some hormones rendering them major targets for the hormonal signaling that regulates spermatogenesis. Although the mechanisms by which SCs fulfill their own and germ cells metabolic needs are mostly studied in vitro, SC metabolism is unquestionably a regulation point for germ cell development and the hormonal control of these processes is required for a normal spermatogenesis.     

The role of the proteasome in the generation of MHC class I ligands and immune responses

The ubiquitin–proteasome system (UPS) degrades intracellular proteins into peptide fragments that can be presented by major histocompatibility complex (MHC) class I molecules. While the UPS is functional in all mammalian cells, its subunit composition differs depending on cell type and stimuli received. Thus, cells of the hematopoietic lineage and cells exposed to (pro)inflammatory cytokines express three proteasome immunosubunits, which form the catalytic centers of immunoproteasomes, and the proteasome activator PA28. Cortical thymic epithelial cells express a thymus-specific proteasome subunit that induces the assembly of thymoproteasomes. We here review new developments regarding the role of these different proteasome components in MHC class I antigen processing, T cell repertoire selection and CD8 T cell responses. We further discuss recently discovered functions of proteasomes in peptide splicing, lymphocyte survival and the regulation of cytokine production and inflammatory responses.     

Germline development in vertebrates and invertebrates

In all animals information is passed from parent to offspring via the germline, which segregates from the soma early in development and undergoes a complex developmental program to give rise to the adult gametes. Many aspects of germline development have been conserved throughout the animal kingdom. Here we review the unique properties of germ cells, the initial determination of germ cell fates, the maintenance of germ cell identity, the migration of germ cells to the somatic gonadal primordia and the proliferation of germ cells during development in vertebrates and invertebrates. Similarities in germline development in such diverse organisms as Drosophila melanogaster, Caenorhabditis elegans, Xenopus laevis and Mus musculus will be highlighted. Received 11 December 1998; received after revision 25 January 1999; accepted 25 January 1999     

Molecular and Cellular Basis of Regeneration and Tissue Repair

Cell plasticity and mesenchymal-epithelial interactions are regarded as a hallmark of embryonic development and are not believed to occur extensively in the adult. Recently, adult mesenchymal stem cells were reported to differentiate in culture into a variety of mature cell types, including epithelial cells. Progress in stem and progenitor cell biology and recognition of the unique properties of such cells may enable intelligent bioengineering design of replacement skin which allows regeneration to occur in vivo. Ideally, a scaffold-free environment which stimulates skin stem cells in situ to initiate cell signals that result in regeneration rather than scar formation is required. Various skin progenitor cell types are considered along with the signalling cascades that they affect. We also discuss a mammalian model of scar-free regeneration. Many of these mechanisms, if fully understood, could be harnessed after injury to perfectly restore the skin.     

The in vivo role of α-mannosidase IIx and its role in processing of <Emphasis Type="Italic">N</Emphasis>-glycans in spermatogenesis

The surfaces of mammalian cells are covered by a variety of carbohydrates linked to proteins and lipids. N-glycans are commonly found carbohydrates in plasma membrane proteins. The structure and biosynthetic pathway of N-glycans have been analyzed extensively. However, functional analysis of cell surface N-glycans is just under way with recent studies of targeted disruption of genes involved in N-glycan synthesis. This review briefly introduces the potential role of processing -mannosidases in N-glycan biosynthesis and recent findings derived from the -mannosidase IIx (MX) gene knockout mouse, which shows male infertility. Thus, the MX gene knockout experiment unveiled a novel function of specific N-glycan, which is N-acetylglucosamine-terminated and fucosylated triantennary structure, in the adhesion between germ cells and Sertoli cells. Analysis of the MX gene knockout mouse is a good example of a multidisciplinary approach leading to a novel discovery in the emerging field of glycobiology.Received 29 November 2002; received after revision 30 December 2002; accepted 20 January 2003     

The germ cell — oncogenic and embryogenic correlates

Summary This study is based on 100 consecutive germ cell tumors of the gonads, received during a 5-year period. Benign teratomas, with totipotent differentiation are the commonest ovarian germ cell tumors, whereas the nullipotent germinomas from the bulk of the testicular tumors. Differentiation in the rapidly proliferating testicular teratomas, occurs in the form of embryoid bodies and organoid structures. From an analysis of the germ cell tumors it is evident that the ovum confers differentiating functions on the zygote, while the spermatozoon confers functions of organization and proliferation. This difference in the behavior of the 2 germ cells is due to local feedbacks within the cortical and medullary zones of the gonads.Acknowledgments. I wish to thank Dr S. K. Lal, Maulana Azad Medical College, New Delhi, for his valuable suggestions concerning gonadal functions. Reprint requests to B. I., B-72, Pandara Road, New Delhi-110003 (India).     

Gene expression in spermiogenesis

Germ cells convey parental genes to the next generation, and only germ cells perform meiosis, which is a mechanism that preserves the parental genes. The fusion of the products of germ cell meiosis, the haploid sperm and egg, creates the next generation. Sperm are the haploid germ cells that contribute genes to the egg. In preparation for this, the haploid round spermatids produced by meiosis undergo drastic morphological changes to become sperm. During this process of spermiogenesis, the nuclear form of the haploid germ cell takes shape, the mitochondria are rearranged in a specific manner, the flagellum develops and the acrosome forms. Spermatogenesis is supported by precise and orderly regulation of gene expression during the changes in chromatin structure, when protamine replaces histone. In this report, we summarize the molecular mechanisms involved in spermiogenesis.Received 2 September 2004; received after revision 7 October 2004; accepted 7 October 2004     

Defending the genome from the enemy within: mechanisms of retrotransposon suppression in the mouse germline

The viability of any species requires that the genome is kept stable as it is transmitted from generation to generation by the germ cells. One of the challenges to transgenerational genome stability is the potential mutagenic activity of transposable genetic elements, particularly retrotransposons. There are many different types of retrotransposon in mammalian genomes, and these target different points in germline development to amplify and integrate into new genomic locations. Germ cells, and their pluripotent developmental precursors, have evolved a variety of genome defence mechanisms that suppress retrotransposon activity and maintain genome stability across the generations. Here, we review recent advances in understanding how retrotransposon activity is suppressed in the mammalian germline, how genes involved in germline genome defence mechanisms are regulated, and the consequences of mutating these genome defence genes for the developing germline.     

Production of transgenic birds

The avian embryo presents a tremendous challenge for those interested in accessing and manipulating the avian germ line. By far the most successful method of gene transfer is by retrovirus vector. The efficacy of retrovirus vectors has been demonstrated by germ line insertion of replication-competent retroviruses as well as the insertion of replication-defective retrovirus vectors carrying bacterial marker genes. Retroviral vectors have also been shown to be useful for the transfer and expression of genes in somatic cells. Further, germ line transgenesis has been reported in both the chicken and the Japanese quail. In addition, several alternative gene transfer methods are under development. These include transfection of avian sperm, development of germ line chimeras using primordial germ cells and blastodermal cells, and the development of embryonic stem cell lines. Potentially, basic research and the poultry industry will derive substantial benefit from this revolutionary technology.     

Production of transgenic birds

The avian embryo presents a tremendous challenge for those interested in accessing and manipulating the avian germ line. By far the most successful method of gene transfer is by retrovirus vector. The efficacy of retrovirus vectors has been demonstrated by germ line insertion of replication-competent retroviruses as well as the insertion of replication-defective retrovirus vectors carrying bacterial marker genes. Retroviral vectors have also been shown to be useful for the transfer and expression of genes in somatic cells. Further, germ line transgenesis has been reported in both the chicken and the Japanese quail. In addition, several alternative gene transfer methods are under development. These include transfection of avian sperm, development of germ line chimeras using primordial germ cells and blastodermal cells, and the development of embryonic stem cell lines. Potentially, basic research and the poultry industry will derive substantial benefit from this revolutionary technology.     

Base excision repair and design of small molecule inhibitors of human DNA polymerase β

Base excision repair (BER) can protect a cell after endogenous or exogenous genotoxic stress, and a deficiency in BER can render a cell hypersensitive to stress-induced apoptotic and necrotic cell death, mutagenesis, and chromosomal rearrangements. However, understanding of the mammalian BER system is not yet complete as it is extraordinarily complex and has many back-up processes that complement a deficiency in any one step. Due of this lack of information, we are unable to make accurate predictions on therapeutic approaches targeting BER. A deeper understanding of BER will eventually allow us to conduct more meaningful clinical interventions. In this review, we will cover historical and recent information on mammalian BER and DNA polymerase β and discuss approaches toward development and use of small molecule inhibitors to manipulate BER. With apologies to others, we will emphasize results obtained in our laboratory and those of our collaborators.     

Glycolipids and phospholipids as natural CD1d-binding NKT cell ligands

Natural killer T (NKT) cells have been shown by a number of studies to play a protective role against cancers, autoimmune diseases and infectious diseases. Several glycolipids and phospholipids derived from mammalian, bacterial, protozoan and plant species have recently been identified as natural ligands (antigens) for NKT cells. Some of these glycolipid/phospholipid ligands have now been crystallized in forms bound to CD1d molecules, and the tertiary structure of these complexes has finally been revealed. This review is intended to list natural NKT cell ligands identified to date, and discuss how their structures relate to their propensity to bind CD1d molecules and, as a consequence, stimulate NKT cells. Received 14 February 2006; received after revision 31 March 2006; accepted 15 May 2006     

Consequences of experimental triploidy on the fertility and sexual differentiation of a normally amphigonic and diploid lombrician, Eisenia foetida Sav]

The fertility of experimental triploid worms has been studied in Eisenia foetida Sav. This study has led us to the conclusion that the female germ line is abortive while the male germ line may as an exception take part in fertilization. Histological controls have confirmed the differentiating ability of the male line and the break in the development of the female line.     

Stem cells and their niche: a matter of fate

Embryonic stem cells provide an in vitro model for developmental biologists to study cell fate decisions during ontogenesis, while somatic stem cells allow physiologists to understand tissue homeostasis in the adult. The behavior of stem cells is dependent on an intimate relationship with a supportive niche. This brief review highlights some of the most important recent trends in stem cell biology, focusing in particular on the supportive microenvironments for both embryonic and adult stem cells. Known intrinsic and extrinsic molecular players from the best-characterized stem cell types are summarized, illuminating a number of shared environmental cues among tissues originating from all three embryonic germ layers. Received 6 October 2005; received after revision 27 December 2005; accepted 17 January 2006