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The trophinin gene encodes a novel group of MAGE proteins, magphinins, and regulates cell proliferation during gametogenesis in the mouse.

Trophinin is a membrane protein that mediates apical cell adhesion between trophoblastic cells and luminal epithelial cells of the endometrium and is implicated in the initial attachment during the process of human embryo implantation. The present study identified novel trophinin gene transcripts, which encode proteins structurally distinct from trophinin protein in the mouse. We designated these proteins "magphinins," because they share consensus amino acid sequences with MAGE (melanoma-associated antigen) superfamily proteins. Among many MAGE proteins, magphinins are closely related to NRAGE, which mediates p75 neurotrophin receptor-dependent apoptosis, and necdin, which is a strong suppressor of cell proliferation in post-mitotic neurons. There are three major forms of magphinins, i.e. magphinin-alpha, -beta, and -gamma, in the mouse, which are formed due to alternative usage of different exons. Northern blot analysis revealed that magphinins are expressed in brain, ovary, testis, and epididymis. In addition, Western blot analysis and in vitro translation experiments showed that magphinins expressed in the mouse ovary and testis are translation products utilizing the second initiation AUG codon and contain an active nuclear localization signal. Ectopic expression of magphinins in mammalian cells resulted in nuclear localization of magphinin and suppressed cell proliferation. Immunohistochemistry of the mouse ovary and testis showed that magphinin proteins are distributed in the cytoplasm of the male and female germ cells, whereas these proteins are translocated to the nucleus at a specific stage of gametogenesis. These results strongly suggest that magphinins regulate cell proliferation during gametogenesis in the mouse.

Alternative Splicing↗

Hsp70 expression and function during gametogenesis.

The dramatic transformations in nuclear content and cellular organization that occur during gametogenesis require unique regulation and execution of the mitotic and meiotic cell cycle, apoptotic cell death, DNA recombination and repair, and cellular differentiation. These processes are accompanied by the constitutive and developmentally regulated expression of a number of hsp70 genes encoding 70 kDa heat shock proteins (Hsp70), including several hsp70s whose expression is unique to male germ cells. Examining the expression and function of Hsp70s in germ cells has provided significant insights into mechanisms of hsp70 gene regulation and Hsp70 protein function, as well as the developmental processes of gametogenesis.

Animals↗

[Histone synthesis in the cells of proliferating and nonproliferating tissues during gametogenesis and early embryogenesis].

A review of available data on the replication-dependent and replication-independent histone synthesis in the proliferating and nonproliferating (quiescent) cells during gametogenesis and early embryogenesis. In each of the considered models the replication-dependent and replication-independent histone synthesis play different roles in the chromatin organization and metabolism. The transition from replication-dependent to replication-independent histone synthesis during gametogenesis is a regular process directed to the formation of a highly compacted metabolically inert chromatin (males) and to the formation of histone protein pool in order to provide the chromatin nucleosome structure in the sperm nucleus during fertilization, as well as the nuclear chromatin in zygotes and blastomeres (females). A suggestion is put forward that the coupling of histone and DNA syntheses should arise not simultaneously in all cells of the embryo but have a regional pattern, due, possibly, to the asynchrony of cell cycle in the early embryos.

Animals↗

[Regularities of radiation effects in ontogenesis of progeny of the 1st generation of Wistar rats after irradiation of germ cells of both parents at different stages of gametogenesis].

The data on antenatal and postnatal periods of ontogenesis of the posterity of the first generation of Wistar rats exposed by doses of 2-4 Gy at various terms before conception showed that the realization of radiation effects depended on the stage of gametogenesis of sex cells of both parents at radiation exposure. Stimulating radiation effect (hyperovulation) is accompanied by the higher rate of antenatal and early postnatal death of the posterity. In the rest cases it died mainly at embryogenesis. The most unfavourable factor for ontogenesis of the posterity appeared to be a participation in the fertilization of sex cells exposed at postmeiotic stages of gametogenesis, with the degree of manifestation of radiation effects depending on radiation dose.

Animals↗

[Gametogenesis and zygote and ookinete formation in some haemoproteid species (Haemosporidia: Haemoproteidae) of birds in vitro].

Observations were made through examination of thin blood films which were prepared at appropriate intervals from citrated fresh blood of infected birds. The comparative description of gametogenesis, zygote and ookinete formation of Haemoproteus tartakovskyi, H. belopolskyi, H. fringillae, H. pallidus under the light microscope is given for the first time. This process has a general pattern for all investigated haemoproteid species. It is shown that macrogametocyte has been transformed directly to macrogamete without forming a residual body, as it was noted by Desser (1972) for H. (= Parahaemoproteus) velans. The most informative signs, which can be used for haemoproteid systematics, were determined. Among them microgamete length, zygote structure, morphological features of developing ookinete as well as the rate of ookinete formation can be pointed out first of all. H. pallidus microgametes are twice shorter than H. tartakovskyi, H. belopolskyi, H. fringillae ones. A large clear vacuole is formed in zygote of H. fringillae only. The majority of diagnostic features has the developing ookinetes. On the basis of ookinetes morphology the investigated species of haemoproteids can be determined easily. There were clear differences between haemoproteid species on the rate of ookinete formation. The ookinetes of H. belopolskyi and H. fringillae are formed for about 12 times more slowly than H. pallidus ones, and for about 6 times more rapidly than H. tartakovskyi ones. The peculiarities of gametogenesis, zygote and ookinete formation under the stable conditions in vitro can be used as usefull additional signs for determination of haemoproteid species.

Animals↗

Gametogenesis processes and multilocus gene identity by descent.

With few exceptions, the determination of unconditional probability of genes shared identical by descent (IBD) by relatives can be very difficult, especially if the relationship is complex or if multiple loci are involved. It is particularly difficult if one needs the IBD probability in a explicit form, expressed in terms of interlocus recombination fractions. In this paper, I will further extend the concept of gametogenesis process introduced elsewhere and indicate that it completely determines the gene IBD events of interest in pedigrees. I will demonstrate that the gametogenesis process not only serves as a convenient conceptual framework in considering IBD events in pedigrees but also provides a simple yet powerful tool to solve a wide range of seemingly difficult problems. In particular, I consider the problem of multilocus IBD probability for relative pairs, k siblings, and a group of pedigree members. In addition, I consider the problem of multilocus autozygosity probability and the problem of gene preservation in close relatives.

Gametogenesis↗

Gene inactivation of Pf11-1 of Plasmodium falciparum by chromosome breakage and healing: identification of a gametocyte-specific protein with a potential role in gametogenesis.

We report the identification of the product of the Plasmodium falciparum Pf11-1 gene and demonstrate that it is a gametocyte-specific protein that has a potential role in the rupture of the host erythrocyte and emergence of the gametes (gametogenesis). The Pf11-1 gene is a large locus (30 kb) whose sequence predicts a glutamic acid-rich polypeptide. Our identification of the Pf11-1 gene product as gametocyte specific was greatly facilitated by the isolation of a mutant parasite clone in which greater than 90% of the Pf11-1 gene was deleted. Molecular analysis of the mutant locus suggests that the underlying genetic mechanism is chromosome breakage and subsequent healing by the addition of telomere repeats. PCR-based analysis showed that similar DNA rearrangements occur commonly in small subpopulations of most laboratory strains, suggesting that the Pf11-1 locus represents a fragile chromosome region. Northern blot analysis demonstrates that a large Pf11-1 gene-specific transcript (much greater than 10 kb) is present in gametocytes but not in asexual blood stage parasites. The Pf11-1 protein was localized by electron microscopy to granules in the cytoplasm of gametocytes adjacent to the membrane of the parasitophorous vacuole. Following in vitro stimulation of gametogenesis, the Pf11-1 protein was found in the membrane of lysed erythrocytes, suggesting a role for Pf11-1 in erythrocyte rupture within the mosquito gut.

Animals↗

Developmental regulation of genomic imprinting during gametogenesis.

Successful mammalian development requires both the male and female genomes. This is due in part to genomic imprinting, which results in offspring inheriting only one functional copy of a gene from either the mother or the father. Evidence suggests that this specialization of the parental genomes is established during gametogenesis when the imprint pattern inherited from the parent is switched to reflect the sex of the progeny. We used reverse transcription-PCR to analyze the allele-specific expression of Igf-2, Igf-2r, and H19 in the testes and ovaries of mice derived from an interspecies cross between Mus musculus and Mus spretus. Because of genomic imprinting, Igf-2 is expressed only from the paternal allele and Igf-2r and H19 only from the maternal allele, in most tissues. Although allele-specific expression was maintained in the neonatal testis and ovary, relaxation of imprinting was detected by 7 days after birth in the male and continued during testis development. In the female, relaxation of the Igf-2 and Igf-2r parental imprints was observed in the adult ovary and oocyte. These results (1) indicate that imprinted expression is relaxed during gametogenesis, presumably as a consequence or prerequisite of the imprinting mechanism, and (2) predict a subsequent imprinting event after which the allele-specific expression of Igf-2, Igf-2r, and H19 reflects the parent of origin.

Alleles↗

The role of X-linked lethal and viable male-sterile mutations in male gametogenesis of Drosophila melanogaster: genetic analysis.

The possibility that viable male-sterile mutations occur in vital genes and the role played by lethal mutations and viable male-steriles in male gametogenesis were studied. Five sterile loci were identified among the 30 most proximal vital loci of the X-chromosome and two of them were shown to be allelic with lethal mutations. Fertility test on gynanders for nonautonomous lethal mutations proved that vital genes operate autonomously in male gonads, independently of their effect on somatic tissues. Fertility tests of ts lethals, shifted to the nonpermissive temperature after the TSP, showed that 40% of vital genes function in male gonads. It is further shown that about the same proportion of vital genes is operating in female gonads and that the two groups overlap by about 70%. The role of viable and lethal male gametogenesis is discussed in detail.

Drosophila melanogaster↗

A genetic locus on Plasmodium falciparum chromosome 12 linked to a defect in mosquito-infectivity and male gametogenesis.

Infection of mosquitoes by Plasmodium spp. requires sexual differentiation of the malarial parasite in the vertebrate host and mating of the heterogametes in the vector midgut. A Plasmodium falciparum clone, Dd2, differentiates into normal-appearing gametocytes, yet poorly infects mosquitoes. The Dd2 clone, however, effectively cross-fertilized HB3, a Central American P. falciparum clone, and yielded several independent recombinant progeny. We have examined 11 HB3 x Dd2 progeny for their ability to infect mosquitoes and to differentiate into male gametes. Our analyses indicate that the poor mosquito-infectivity of the Dd2 clone results from a defect in male gametogenesis. This defect was inherited as a single locus in the independent recombinant progeny of HB3 x Dd2. Comparison with a restriction fragment length polymorphism map of the HB3 x Dd2 cross indicates that the defective phenotype of Dd2 maps to a locus on P. falciparum chromosome 12. This genetic locus may contain determinants that play a crucial role in male gametogenesis by P. falciparum.

Animals↗

The ubiquitin system in gametogenesis.

Ubiquitin is a ubiquitous and highly conserved protein of 76 amino acid residues, that can be covalently attached to cellular acceptor proteins. The attachment of ubiquitin to target proteins is achieved through a multi-step enzymatic pathway, which involves activities of ubiquitin-activating E1 enzymes, ubiquitin-conjugating E2 enzymes, and ligating E3 enzymes. Mono- or poly-ubiquitination of proteins can lead to protein degradation or modification of protein activity. Many components of the complex ubiquitin system show remarkable evolutionary conservation, from yeast to mammalian species. The ubiquitin system is essential to all eukaryotic cells. Among others, several signal transduction cascades show involvement of the ubiquitin system, but there are currently little data supporting a specific role of the ubiquitin system in hormonal control of reproduction. Interestingly, during gametogenesis, many specialized and important aspects of the ubiquitin system become apparent. Components of the ubiquitin system appear to be involved in different steps and processes during gametogenesis, including control of meiosis, and reorganization of chromatin structure.

Animals↗

Selection for conditional gametogenesis in Chlamydomonas reinhardi.

A new technique is described for selection of temperature-sensitive mutants affecting gametogenesis in Chlamydomonas reinhardi. The first mutant found by this technique is characterized. Cells exhibiting the gam-1 phenotype are capable of sexual agglutination, but cannot form zygotes at the restrictive temperature. The mutation, however, has been expressed only in gametes of mating type (-). Cells of mating type (+) which carry this gene are able to engage in normal zygote formation. Temperature shift experiments and antibiotic studies have indicated that this gene is expressed within 6 hr after the onset of gametogenesis in liquid culture, and its product has a lifetime of about 4 hr at 35 degrees.

Chlamydomonas↗

SLOW WALKER1, essential for gametogenesis in Arabidopsis, encodes a WD40 protein involved in 18S ribosomal RNA biogenesis.

The progression of mitotic division cycles and synchronous development between and within the male and female reproductive organs are essential for plant sexual reproduction. Little is known about the genetic control of the progression of mitotic cycles of the haploid genome during gametogenesis in higher plants. Here, we report the phenotypic and molecular characterization of an Arabidopsis thaliana mutant, slow walker1 (swa1), in which the progression of the mitotic division cycles of the female gametophyte was disrupted. Confocal microscopy revealed that megagametophyte development was asynchronous in swa1, causing embryo sacs to arrest at two-, four-, or eight-nucleate stages within the same pistil. A delayed pollination experiment showed that a small fraction of the swa1 embryo sacs were able to develop into functional female gametophytes. The swa1 mutation also showed a slight reduction in penetrance through the male gametophyte, although the pollen grains were morphologically normal. Molecular analysis indicates that SWA1 encodes a protein with six WD40 repeats that is localized in the nucleolus in interphase cells. The SWA1 gene is expressed in cells undergoing active cell divisions, including functional megaspores and the female gametophytic cells. RNA interference results indicated that knockout of SWA1 inhibited root growth significantly and led to the accumulation of unprocessed 18S pre-rRNA. These data suggest that SWA1 most likely plays a role in rRNA biogenesis that is essential for the progression of the mitotic division cycles during gametogenesis in plants.

Amino Acid Sequence↗

Requirement of aminoacyl-tRNA synthetases for gametogenesis and embryo development in Arabidopsis.

Aminoacyl-tRNA synthetases (AARSs) are required for translation in three different compartments of the plant cell: chloroplasts, mitochondria and the cytosol. Elimination of this basal function should result in lethality early in development. Phenotypes of individual mutants may vary considerably, depending on patterns of gene expression, functional redundancy, allele strength and protein localization. We describe here a reverse genetic screen of 50 insertion mutants disrupted in 21 of the 45 predicted AARSs in Arabidopsis. Our initial goal was to find additional EMB genes with a loss-of-function phenotype in the seed. Several different classes of knockouts were discovered, with defects in both gametogenesis and seed development. Three major trends were observed. Disruption of translation in chloroplasts often results in seed abortion at the transition stage of embryogenesis with minimal effects on gametophytes. Disruption of translation in mitochondria often results in ovule abortion before and immediately after fertilization. This early phenotype was frequently missed in prior screens for embryo-defective mutants. Knockout alleles of non-redundant cytosolic AARSs were in general not identified, consistent with the absolute requirement of cytosolic translation for development of male and female gametophytes. These results provide a framework for evaluating redundant functions of AARSs in Arabidopsis, a valuable data set of phenotypes resulting from multiple disruptions of a single basal process, and insights into which genes are required for both gametogenesis and embryo development and might therefore escape detection in screens for embryo-defective mutants.

Amino Acyl-tRNA Synthetases↗

New observations on gametogenesis, fertilization, and zygote transformation in Plasmodium gallinaceum.

The ultrastructure of the sexual stages of Plasmodium gallinaceum during gametogenesis, fertilization, and early zygote transformation is described. New observations are made regarding the parasitophorous vacuole (PV) of gametocytes and the process of emergence in male and female gametocytes. Whereas female gametocytes readily disrupted both the PV membrane and host cell plasmalemma during emergence, male gametocytes frequently failed to break down the plasmalemma of the host cell. New observations and hypotheses are presented on the behavior of the male gamete nucleus. Following fertilization, the male nucleus appears to travel through a channel of endoplasmic reticulum in the female gamete before fusing with the female nucleus at a region in which the nuclear envelope is thrown into extensive convoluted folds. Polarization of the zygote nucleus, in association with the appearance of a perinuclear spindle of cytoplasmic microtubules, preceded all other changes in the developing zygote. After nuclear polarization becomes apparent, electron-dense material is deposited beneath the zygote pellicle, and a canopy is formed which eventually extends over the entire apical end of the developing ookinete. As the apical end begins to extend outward, polar rings, micronemes, and subpellicular microtubules become visible in this portion and a "virus-like" inclusion known as a crystalloid is formed in the posterior portion of the zygote. When female gametes are prevented from being fertilized, the cytoplasm at 24h after gametogenesis is devoid of most of those organelles found in the developing zygote or the mature ookinete. The cell is surrounded only by a single membrane. Although at various points beneath the membrane there are deposits of electron-dense material reminiscent of those deposited in the zygote, no further development of ookinete structures takes place in the unfertilized female gamete.

Animals↗

Gametogenesis, fertilization and ookinete differentiation of Leucocytozoon smithi.

Development of Leucocytozoon smithi during gametogenesis, fertilization, and ookinete differentiation was studied by light and electron microscopy. Gametogenesis occurred rapidly, within 1-2 min after gametocytes were ingested by black flies. Usually one axoneme, but not infrequently two, was observed in microgametes. The macrogamete nucleus was characteristically elongated and fragmented, with a convoluted nuclear envelope. Fertilization occurred within five min after ingestion of gametocytes by the vector. The entire axoneme and nucleus of the microgamete entered the cytoplasm of the macrogamete. Zygote differentiation resembled sporozoite formation in that a thickened inner membrane and subpellicular microtubules developed beneath the plasmalemma, followed by cytoplasmic protrusion or evagination to form the anterior end. Extension of the inner thickened membrane continued as the zygote elongated. Development of sausage-shaped ookinetes was completed within 6-8 h after ingestion of a blood meal by a black fly. Mature ookinetes possessed a single nucleus, double-layered pellicle, canopy, apical pore, polar ring complex, subpellicular microtubules, micronemes, crystalloids, abundant mitochondria, endoplasmic reticulum, and ribosomes. Comparison of development of L. smithi with species of Plasmodium and Haemoproteus revealed general similarities in both sexual and asexual development within the insect vector. A diagram summarizing life cycle events for L. smithi is included.

Animals↗

Developmentally regulated expression of cyclin D3 and its potential in vivo interacting proteins during murine gametogenesis.

To begin to assess the function of the cell cycle regulator cyclin D3 during gametogenesis, the present study examined its expression, interacting partners, and associated kinase activity in the murine testis and ovary. In the early stages of postnatal testicular development, cyclin D3 protein was detected in spermatogonia and Leydig cells. In the adult testis, cyclin D3 was also expressed in terminally differentiating spermatids. In the embryonic ovary, detection of cyclin D3 was limited to somatic cells. In the postnatal ovary, its localization was predominantly in the nuclei of oocytes in primordial and small follicles, a localization that diminished with oocyte growth. Cdk4 and p27 were expressed in a similar subset of testicular and ovarian cells, suggesting that they may regulate cyclin D3 function during testicular and ovarian development in a cell type-specific manner. Cyclin D3-associated kinase activity was detected in immature, but not adult, testes and ovaries. These observations suggest unique roles for cyclin D3 in the control of cell division and differentiation in the germ line and the differential regulation of mitotic and meiotic cell cycles during male and female gametogenesis.

Animals↗

Kinetics of the male gametogenesis.

The kinetics of the male gametogenesis during the pregonadal period, prespermatogenesis, and "early" spermatogenesis has been described in detail. Concerning spermatogenesis in the adult individual reference is made to the articles of Courot, Hochereau-de Reviers and Ortavant (1970) and of Clermont (1972). The comparison of female and male gametogenesis (Fig. 1) shows that the "gonia stage" (asterisks) of the female germ cells is limited to one proliferation wave only, whereas the "gonia stage" of the male germ cells consists of a first proliferation wave, comparable to that of oogonia, a preparative phase to initiate spermatogenesis, and a second proliferation wave with renewal and differentiation of the spermatogonia. Germ cells in the "gonia stage" are highly sensitive towards ionising radiation and cytostatic drugs.

Animals↗