Gamete development encompasses key cellular and molecular processes common to gametes of both sexes as well as sex-specific processes crucial for fertilization and embryogenesis. Common gametogenic pathways include meiosis, to carry out recombination and segregation of homologous chromosomes, and silencing of retrotransposons, to maintain genomic integrity. Precise control of these processes is essential for transmission of parental genetic materials to subsequent generations and creation of genetic diversity. Errors can cause infertility, miscarriage, or birth defects and pose genetic threats to the offspring and subsequent generations. Here we put forward the hypothesis that meiosis arrest female 1 (MARF1) expressed in mouse oocytes is a functional counterpart of the nuage-associated components of spermatocytes that carry out these crucial facets of regulation of meiosis and control of retrotransposon expression.

In April 2008, outside of Cambridge, England, The Hinxton Group convened an international consortium to discuss an exciting new area of research that has emerged over the last few years: the production of germ cells from pluripotent stem cells in vitro. Although functional eggs or sperm have not yet been produced in vitro from either embryonic stem (ES) cells or induced pluripotent stem (iPS) cells, progress has been tantalizing. This research opens new opportunities for studies on the development of germ cells and prospects for clinical applications. The goal of the Hinxton Group was to provide conceptual framework and guiding principles for policy makers, and to this end, ‘‘A Consensus Statement: Science, Ethics and Policy Challenges of Pluripotent Stem Cell-Derived Gametes’’ emerged after three days of discussion and debate. This statement, reproduced here verbatim, should stimulate interest and discussion throughout the community of reproductive and developmental biologists and provide a framework for societal debate.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Abstract The objective of this study was to improve the conditions for oocyte development in vitro beginning with the primordial follicles of newborn mice. Previous studies showed that oocytes competent of meiotic maturation, fertilization, and preimplantation could develop in vitro from primordial follicles. However, the success rates were low and only one live offspring was produced (0.5% of embryos transferred). A revised protocol was compared with the original protocol using oocyte maturation and preimplantation development as end points. The percentage of oocytes maturing to metaphase II and developing to the blastocyst stage was significantly improved using the revised protocol. In addition, we compared the production of offspring from two-cell stage embryos derived from in vitro-grown and in vivo-grown oocytes. Of 1160 transferred two-cell stage embryos derived from in vitro-grown oocytes, 66 (5.7%) developed to term and 7 pups (10.6%) died at birth. The remaining 59 pups (27 females, 32 males) survived to adulthood. By comparison, of 437 transferred two-cell stage embryos derived from in vivo-grown oocytes, 76 (17.4%) developed to term and 4 (5.3%) died at birth. The remaining 72 pups (35 females, 37 males) survived to adulthood. These studies provide proof of the principle that fully competent mammalian oocytes can develop in vitro from primordial follicles and present a significant advance in oocyte culture technology.

All vertebrate eggs are surrounded by an extracellular matrix. This matrix is known as the zona pellucida in mammals and is critically important for the survival of growing oocytes, successful fertilization and the passage of early embryos through the oviduct, The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2 and ZP3), each encoded by a single copy gene, Using targeted mutagenesis in embryonic stem cells, Zp2-null mouse lines have been established, ZP1 and ZP3 proteins continue to be synthesized and form a thin zona matrix in early follicles that is not sustained in pre-ovulatory follicles, The abnormal zona matrix does not affect initial folliculogenesis, but there is a significant decrease in the number of antral stage follicles in ovaries isolated from mice lacking a zona pellucida, Few eggs are detected in the oviduct after stimulation with gonadotropins, and no two-cell embryos are recovered after mating Zp2-null females with normal male mice. The structural defect is more severe than that observed in Zp1-null mice, which have decreased fecundity, but not quite as severe as that observed in Zp3-null mice, which never form a visible zona pellucida and are sterile. Although zona-free oocytes matured and fertilized in vitro can progress to the blastocyst stage, the developmental potential of blastocysts derived from either Zp2- or Zp3-null eggs appears compromised and, after transfer to foster mothers, live births have not been observed. Thus, in addition to its role in fertilization and protection of early embryos, these data are consistent with the zona pellucida maintaining interactions between granulosa cells and oocytes during folliculogenesis that are critical to maximize developmental competence of oocytes.

With the reports of the DNA sequence of the human and mouse genomes, research attention is now on questions of protein function and interactions, developmental and physiological pathways, and systems biology. The International Mouse Mutagenesis Consortium proposes goals and outlines plans for the next 10 years for annotating the mouse genome and compiling data on mouse mutations. In addition to the challenges of identifying new assays to probe biological functions, more efficient and reliable methods are needed for archiving, managing, analyzing, displaying, and disseminating the complex phenotype data sets resulting from mutagenesis programs. Achieving these goals will require the biomedical research community to improve efficiencies, to reduce costs, and to coordinate international expertise and resources.

Little is known about gene action in the preimplantation events that initiate mammalian development. Based on cDNA collections made from each stage from egg to blastocyst, 25438 3'-ESTs were derived, and represent 9718 genes, half of them novel. Thus, a considerable fraction of mammalian genes is dedicated to embryonic expression. This study reveals profound changes in gene expression that include the transient induction of transcripts at each stage. These results raise the possibility that development is driven by the action of a series of stage-specific expressed genes. The new genes, 798 of them placed on the mouse genetic map, provide entry points for analyses of human and mouse developmental disorders.

Mammalian embryos can only survive if they attach to the uterus (implantation) and establish proper maternal-fetal interactions. To understand this complex implantation pathway, we have initiated genomic analysis with a systematic study of the cohort of genes expressed in extraembryonic cells that are derived from the conceptus and play a major role in this process. A total of 2103 cDNAs from the extraembryonic portion of 7.5-day post-conception mouse embryos yielded 3186 expressed sequence tags, approximately 40% of which were novel to the sequence databases. Furthermore, when 155 of the cDNA clones with no homology to previously detected genes were genetically mapped, apparent clustering of these expressed genes was detected in subregions of chromosomes 2, 7, 9 and 17, with 6.5% of the observed genes localized in the t-complex region of chromosome 17, which represents only approximately 1.5% of the mouse genome. In contrast, X-linked genes were under-represented. Semi-quantitative RT-PCR analyses of the mapped genes demonstrated that one third of the genes were expressed solely in extraembryonic tissue and an additional one third of the genes were expressed predominantly in the extraembryonic tissues. The over-representation of extraembryonic-expressed genes in dosage-sensitive autosomal imprinted regions and under-representation on the dosage-compensated X chromosome may reflect a need for tight quantitative control of expression during development.

We have demonstrated previously that noncoding sequences of genes are a robust source of polymorphisms between mouse species when tested using single-strand conformation polymorphism (SSCP) analysis, and that these polymorphisms are useful for genetic mapping. In this report we demonstrate that presumptive 3'-untranslated region sequence obtained from expressed sequence tags (ESTs) can be analyzed in a similar fashion, and we have used this approach to map 262 loci using an interspecific backcross. These results demonstrate SSCP analysis of genes or ESTs is a simple and efficient means for the genetic localization of transcribed sequences, and is furthermore an approach that is applicable to any system for which there is sufficient sequence polymorphism.

A system is described here by which live mice can be produced from oocytes isolated from 12-day-old mice, be grown, matured, and fertilized in vitro, and then be transferred to pseudopregnant females. These oocytes were, at the time of isolation from preantral follicles, in about mid-growth phase and incompetent of undergoing germinal vesicle breakdown (GVB) without further development. The developmental competence of mouse oocytes that grew and underwent maturation in vitro was compared to oocytes that grew in vivo and underwent maturation in vitro. After isolation from mice 16 through 28 days old, oocytes were found to increase in size and to sequentially acquire the ability to undergo GVB, produce a polar body, cleave to the 2-cell stage after insemination, and develop to the blastocyst stage. Moreover, the number of cells per blastocyst increased with the age of the mice from which the immature oocytes were isolated. Oocyte-granulosa cell complexes isolated from 12-day-old mice were cultured for 10 days. At the end of the culture period, the oocytes had grown to a size equivalent to oocytes isolated from 16-day-old mice, and 87% of the in-vitro-grown (IVG) oocytes underwent GVB; 79% of these produced a clearly visible polar body when maturation occurred in the presence of follicle-stimulating hormone (FSH). The IVG oocytes cleaved to the 2-cell stage after insemination in vitro with a frequency equivalent to superovulated ova and ova that matured in vitro after isolation from 22-day-old mice.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between cAMP and the capacity of two partially-purified fractions of porcine follicular fluid (PFF) to suppress mouse oocyte maturation was investigated. The fractions used were: 1) a low molecular weight filtrate (less than 10,000) derived from Amicon PM10 filtration of PFF (PM10), and 2) a fraction from Bio-Gel P2 chromatography of the PM10 filtrate (Bio-Gel) that has meiotic inhibitory activity in porcine oocytes. PM10 alone produced a transient inhibition of the maturation of both cumulus cell-enclosed and denuded mouse oocytes, as manifested by germinal vesicle breakdown. The addition of FSH or (Bu)2cAMP to medium containing either of the PFF fractions resulted in dramatic synergism of the inhibitory effect of PFF alone in cumulus cell-enclosed oocytes. Forskolin or (Bu)2cAMP promoted a similar synergistic response with either PFF fraction in denuded oocytes. The degree of inhibition was consistently greater in cumulus cell-enclosed than in denuded oocytes. The putative inhibitor was dialyzable through tubing having a nominal molecular weight cutoff of 1,000. Proteolysis, acid hydrolysis, or ether extraction of PM10 did not reduce its inhibitory synergism with (Bu)2cAMP. However, inhibition was completely abolished by charcoal extraction. Steroid hormones did not mimic the PM10-induced synergism when added to (Bu)2cAMP-containing medium in a concentration at least 14-fold greater than that present in PM10-supplemented medium. In conclusion, our results demonstrate the presence of a factor(s) in PFF that acts synergistically with a cAMP-dependent process to inhibit oocyte maturation in vitro. Furthermore, although PFF fractions suppressed the maturation of both denuded and cumulus cell-enclosed oocytes, cumulus cells appear to mediate the inhibitory activity of PFF. The data also suggest that the PFF inhibitor is a small (mol wt less than 1,000) hydrophobic molecule but not a peptide or nonpolar lipid.

Des granules de mélanine furent extraites des yeux d'embryons de poussins et de souris noires venant de naître ainsi que d'œufs deXenopus laevis et du mélanome S-91. Après que des purifications extensives de granules de mélanine furent mises en solution soit dans l'urée de 8 M ou 1% de SDS et caractérisées par électrophorèse en gel de polyacrylamide. Le résultat indique que plusieurs protéines de même comportement en électrophorèse son présents dans les granules de ces diverses provenances. En plus, il semble encore plus significatif qu'il y ait plusieurs différences entre les protéines mélanosomales des mélanocytes normaux et nocifs.