Genes associated with the development of the male germ line

The development of the mammalian germ line has been well studied, from the designation of primordial germ cells and their migration in the embryo to their progression through gametogenesis. The pattern of germ cell development, as established through classical studies, is now being overlaid with molecular, genetic and epigenetic data. Eventually, proteonomics will lead to a deeper understanding of the function of these genes. Through knowledge of germ cell gene expression patterns, it is now possible to develop transgenic molecular tools for the isolation of germ cells at different stages of development. By linking stage-specific germ cell promoter regions to the green fluorescent protein (GFP) reporter gene it is possible to tag these cells genetically for histological identification and cell sorting. Our long-term goal is to develop male germ cells as stem cells for therapeutic purposes. It is hoped that this goal will be achieved by purifying germ cells at different stages in development and gaining a deeper understanding of them by studying their gene expression patterns, potency and plasticity, both in vivo and in vitro.     

Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease

The human male is characterized by extremely poor semen quality as reflected in the number, morphology and motility of the spermatozoa and a high incidence of nuclear and mitochondrial DNA damage. As a consequence of these factors, defective sperm function is thought to be a major contributor to the aetiology of human infertility, as well as childhood diseases including dominant genetic mutations such as achondroplasia and cancer. Factors associated with the origin of poor semen quality include: (i) a lack of selection pressure for high fecundity genes in developed countries, (ii) an evolutionary lineage associated with the deterioration of several male fertility genes in humans and their close ancestors, (iii) genetic factors including, but not limited to, Y-chromosome deletions (iv) paternal age and (v) environmental factors. A model is proposed whereby factors such as ageing or environmental toxicants initiate DNA strand breakage in the spermatozoa of affected males, eventually leading to a mutation in the embryo. This hypothesis stresses the importance of discovering the identity of those environmental factors that are capable of damaging DNA integrity in the male germ line. Such information could make an important contribution to understanding of the origins of both male infertility and a variety of pathological conditions that affect humans, including cancer and dominant genetic disease.     

Molecular biological features of male germ cell differentiation

Somatic cell differentiation is required throughout the life of a multicellular organism to maintain homeostasis. In contrast, germ cells have only one specific function; to preserve the species by conveying the parental genes to the next generation. Recent studies of the development and molecular biology of the male germ cell have identified many genes, or isoforms, that are specifically expressed in the male germ cell. In the present review, we consider the unique features of male germ cell differentiation. (Reprod Med Biol 2007; 6 : 1–9)     

Origins and consequences of DNA damage in male germ cells

DNA damage in the male germline is associated with poor fertilization rates following IVF, defective preimplantation embryonic development, and high rates of miscarriage and morbidity in the offspring, including childhood cancer. This damage is poorly characterized, but is known to involve hypomethylation of key genes, oxidative base damage, endonuclease-mediated cleavage and the formation of adducts with xenobiotics and the products of lipid peroxidation. There are many possible causes of such DNA damage, including abortive apoptosis, the oxidative stress associated with male genital tract infection, exposure to redox cycling chemicals, and defects of spermiogenesis associated with the retention of excess residual cytoplasm. Physical factors such as exposure to radiofrequency electromagnetic radiation or mild scrotal heating can also induce DNA damage in mammalian spermatozoa, although the underlying mechanisms are unclear. Ultimately, resolving the precise nature of the DNA lesions present in the spermatozoa of infertile men will be an important step towards uncovering the aetiology of this damage and developing strategies for its clinical management.     

Hormonal and genetic control of germ cell apoptosis in the testis

Programmed cell death is an evolutionarily conserved cell death process that plays a major role during normal development and homeostasis. In many cases, the ordered execution of this internal death programme leads to typical morphological and biochemical changes that have been termed apoptosis. The crucial role of this mode of cell death in the pathogenesis of diverse human diseases including cancer, acquired immunodeficiency syndrome, neurodegeneratives disorders, atherosclerosis and cardiomyopathy is now supported by a wealth of data. In adult mammals, including humans, germ cell death is conspicuous during normal spermatogenesis and plays a pivotal role in sperm output. Withdrawal of gonadotrophins and testosterone further enhances the degeneration of germ cells in the testis. The availability of a quantitative method for analysing the testicular DNA fragmentation and in situ methods to localize specific germ cells undergoing apoptosis, either spontaneously or in response to a variety of death triggering signals, opens new avenues in the understanding of the significance of germ cell apoptosis during normal and abnormal states of spermatogenesis. A growing body of evidence demonstrates that both spontaneous (during normal spermatogenesis) and accelerated germ cell death triggered by deprivation of the gonadotrophic support or moderately increased scrotal temperature in adult rats occur almost exclusively via apoptosis. Although there has been spectacular progress in the understanding of the molecular mechanisms of apoptosis in various systems other than spermatogenesis, elucidation of the biochemical and molecular mechanisms by which germ cell apoptosis is regulated has only just begun. It is likely that germ cell apoptosis is controlled in a cell-type specific fashion, but the basic elements of the death machinery may be universal. In addition, there is increasing evidence that homozygous disruption of a number of genes in mice results in infertility through accelerated germ cell apoptosis. Manipulation of spermatogenesis by survival factor(s) deprivation or increases in extrinsic death signals in loss-of-function or gain-of-function mouse models provides a basis for further attempts to define the intrinsic regulation of various death-related genes by external death signals. Such information is crucial for effective management of male factor infertility as well as more targeted approaches to male contraception.     

Fertilization of mouse oocytes using somatic cells as male germ cells

Female and male mouse somatic cells were injected into mouse F(1) oocytes. The cells used included cumulus cells (female) and muscle derived fibroblasts (male). The ability of the cells to fertilize oocytes and support embryonic development was examined. Following activation of the injected oocytes, two second polar bodies were extruded and two pronuclei were formed, one derived from the oocyte chromosomes and the other from the somatic cell chromosomes in a similar way to that observed following fertilization with secondary spermatocytes. Both second polar bodies contained DNA. The fertilization rates by cumulus cells were 10-29%. This was dependent on the artificial activation protocol and on the age of the oocytes. Older oocytes recovered 16-17 h after human chorionic gonadotrophin (HCG) injection were more likely to produce two second polar bodies and two pronuclei than young oocytes which were retrieved at 13-14 h after HCG injection (P < 0.01). The fertilization rates with fibroblasts were 29% using the most effective activation regime and aged oocytes. Most (80-90%) of the 'zygotes' produced by somatic cells cleaved to two cells in culture and ~50% reached the morula stage. However, the developmental competence of the embryos to reach blastocysts was limited. The present study demonstrates that mouse somatic cells undergo haploidization when injected into metaphase II oocytes, fertilize oocytes as diploid male germ cells and support preimplantation development to a degree.     

Transient receptor potential vanilloid receptor-1 confers heat resistance to male germ cells

Testicular hyperthermia in mice lacking transient receptor potential vanilloid receptor-1 results in a much more rapid and massive germ cell depletion from the seminiferous tubules than in wild-type animals, indicating that this receptor protects germ cells against heat stress.     

Antihormones and control of male fertility

Androgens play a very important role at every level in spermatogenesis; it is, therefore, extremely difficult to modify male fertility without imparing sexual behavior. This article reviews different studies conducted with several types of antihormones utilized in rats, hamsters, and rabbits. Agents used were gonadotropin-releasing hormone agonists, antigonadotropins, antiprolactinemic agents, and antiandrogens. It appears that, among the several antihormones, only antifollicle-stimulating hormone do not modify testosteronemia, and would be the ideal spermatogenesis-blocking agents. However, since spermatogenesis is also under the control of the Sertoli cells, future studies should investigate Sertolian secretion, and its influence on epididymal maturation.     

Epigenetic programming of the germ line: effects of endocrine disruptors on the development of transgenerational disease

Epigenetic programming of the germ line occurs during embryonic development in a sex-specific manner. The male germ line becomes imprinted following sex determination. Environmental influences can alter this epigenetic programming and affect not only the developing offspring, but also potentially subsequent generations. Exposure to an endocrine disruptor (i.e. vinclozolin) during embryonic gonadal sex determination can alter the male germ-line epigenetics (e.g. DNA methylation). The epigenetic mechanism involves the alteration of DNA methylation in the germ line that appears to transmit transgenerational adult onset disease, including spermatogenic defects, prostate disease, kidney disease and cancer.     

Co-treatment of the male partner in vaginal candidosis: a double-blind randomized control study

In a double-blind study, 117 non-pregnant women with vaginal candidosis were treated for 3 days with 200 mg-tablets of ketoconazole taken once, twice or three times daily. The incidence of predisposing factors or of a recurrence history did not differ between treatment groups. Their male partners were randomly assigned to receive ketoconazole, 200 mg twice daily or placebo for 3 consecutive days. Cure and recurrence rates were not different in the three treatment groups, with or without simultaneous treatment of the male partner. Treatment of the sexual partner in the three dose-regimen-groups proved not to have influenced the therapeutic effect.     

Co-treatment of the male partner in vaginal candidosis: a double-blind randomized control study

Summary. In a double-blind study, 117 non-pregnant women with vaginal candidosis were treated for 3 days with 200 mg-tablets of ket-oconazole taken once, twice or three times daily. The incidence of predisposing factors or of a recurrence history did not differ between treatment groups. Their male partners were randomly assigned to receive ketoconazole, 200 mg twice daily or placebo for 3 consecutive days. Cure and recurrence rates were not different in the three treatment groups, with or without simultaneous treatment of the male partner. Treatment of the sexual partner in the three dose-regimengroups proved not to have influenced the therapeutic effect.