Diurnal dynamics of S-phase entry of germ cells in the secondary testis of the bambooleaf wrasse (Pseudolabrus sieboldi)

The diversity of reproductive strategies found in fish is attributed to the adaptation of gametogenesis to different habitats. To date, however, information about the dynamics of male gametogenesis and its molecular mechanisms of control by the brain-pituitary-gonadal (BPG) axis, a well-known facet of the endocrine system in vertebrates, is not sufficient to explain the variation in spawning and regulatory mechanisms among species. The bambooleaf wrasse (Pseudolabrus sieboldi) is a protogynous hermaphrodite fish that shows clear diurnal fluctuations in gonadotropin gene expression and serum sex steroid levels associated with spawning. In this study, morphometrical and histological analyses were performed to determine the number of spermatogonial generations in the testis of the sex-changed male (secondary testis). In addition, the diurnal dynamics of S-phase entry of germ cells was explored by measuring the frequency of BrdU-incorporating germ cells at different times of day. We found that the bambooleaf wrasse spermatozoa were generated through nine spermatogonial generations, followed by meiotic divisions accompanied by loss of some spermatocytes and spermatids through apoptosis. BrdU analyses revealed a high frequency of entry into S-phase of undifferentiated type A spermatogonia at 03:00?hr. On the other hand, the frequency of spermatocytes at S-phase of the cell cycle decreased during 00:00-09:00?hr. This study demonstrates for the first time the daily fluctuations of S-phase entry of male germ cells in fish. These results will provide a useful foundation for understanding the roles of endocrinological and cytological regulation of fish spermatogenesis.     

Implications of epigenetics and genomic imprinting in assisted reproductive technologies

There have been several reports of an increased risk of genomic imprinting disorders associated with assisted reproductive technology (ART). However, the connection between imprinting defects and ART is tenuous. In this review, this putative association is investigated in detail, with emphasis on particular steps of the ART process, and which of these may be prone to induction of imprinting errors due to synchrony with major imprinting events during gametogenesis, fertilization, and early embryonic development. While contributions from in vitro manipulation of gametes and embryos cannot be ruled out, it appears that superovulation and/or the condition of infertility, itself, may be largely responsible for the increased risk of genomic imprinting disorders observed with ART births. However, two significant shortcomings of all of these studies preclude rigorous exploration of this issue: the lack of large, longitudinal studies on specific cohorts of ART‐conceived children, and questions surrounding the primary generation of expression and epigenetic data from oocytes and embryos. Future directions for study are proposed, along with a preview of what may be in store as the art of ART advances. Mol. Reprod. Dev. 76: 1006–1018, 2009. © 2009 Wiley‐Liss, Inc.     

Widespread Presence of Human BOULE Homologs among Animals and Conservation of Their Ancient Reproductive Function

Sex-specific traits that lead to the production of dimorphic gametes, sperm in males and eggs in females, are fundamental for sexual reproduction and accordingly widespread among animals. Yet the sex-biased genes that underlie these sex-specific traits are under strong selective pressure, and as a result of adaptive evolution they often become divergent. Indeed out of hundreds of male or female fertility genes identified in diverse organisms, only a very small number of them are implicated specifically in reproduction in more than one lineage. Few genes have exhibited a sex-biased, reproductive-specific requirement beyond a given phylum, raising the question of whether any sex-specific gametogenesis factors could be conserved and whether gametogenesis might have evolved multiple times. Here we describe a metazoan origin of a conserved human reproductive protein, BOULE, and its prevalence from primitive basal metazoans to chordates. We found that BOULE homologs are present in the genomes of representative species of each of the major lineages of metazoans and exhibit reproductive-specific expression in all species examined, with a preponderance of male-biased expression. Examination of Boule evolution within insect and mammalian lineages revealed little evidence for accelerated evolution, unlike most reproductive genes. Instead, purifying selection was the major force behind Boule evolution. Furthermore, loss of function of mammalian Boule resulted in male-specific infertility and a global arrest of sperm development remarkably similar to the phenotype in an insect boule mutation. This work demonstrates the conservation of a reproductive protein throughout eumetazoa, its predominant testis-biased expression in diverse bilaterian species, and conservation of a male gametogenic requirement in mice. This shows an ancient gametogenesis requirement for Boule among Bilateria and supports a model of a common origin of spermatogenesis.     

Ice age: Cryopreservation in assisted reproduction – An update

Since the first reported birth following in vitro fertilization in 1978, further developments in assisted reproductive technology (ART) treatments have produced at least 8 million babies worldwide. Cryopreservation techniques have been central to this treatment revolution, increasing cycle efficacy by allowing the banking of supernumerary embryos for later use, as well as affording the prospective patient more time in cases of anticipated fertility decline. Additionally, these techniques have demonstrated promise in increasing the safety of ART treatments, by reducing complications such as ovarian hyperstimulation, leading to increased support for the introduction of a ‘total freeze’ policy involving deferred embryo transfers. Importantly, the effective cryopreservation of both spermatozoa and oocytes has permitted long-term gamete storage without degradation of quality, facilitating gamete banking for personal use or fertility treatment. Here, we will summarise the indications for applying cryopreservation methods in clinical reproductive medicine, highlighting recent technical advances and examining the evidence base that supports the continued use of cryopreservation in ART.     

Research in nondomestic species: experiences in reproductive physiology research for conservation of endangered felids

Tremendous strides have been made in recent years to broaden our understanding of reproductive processes in nondomestic felid species and further our capacity to use this basic knowledge to control and manipulate reproduction of endangered cats. Much of that progress has culminated from detailed scientific studies conducted in nontraditional laboratory settings, frequently at collaborating zoological parks but also under more primitive conditions, including in the field. A mobile laboratory approach is described, which incorporates a diverse array of disciplines and research techniques. This approach has been extremely useful, especially for conducting gamete characterization and function studies as well as reproductive surveys, and for facilitating the development of assisted reproductive technology. With continuing advances in assisted reproduction in rare felids, more procedures are being conducted primarily as service-related activities, targeted to increase effectiveness of species propagation and population management. It can be a challenge for both investigators and institutional animal care and use committees (IACUCs) to differentiate these service-based procedures from traditional research studies (that require IACUC oversight). For research with rare cat species, multi-institutional collaboration frequently is necessary to gain access to scientifically meaningful numbers of study subjects. Similarly, for service-based efforts, the ability to perform reproductive procedures across institutions under nonstandard laboratory conditions is critical to applying reproductive sciences for managing and preserving threatened cat populations. Reproductive sciences can most effectively assist population management programs (e.g., Species Survival Plans) in addressing conservation priorities if these research and service-related procedures can be conducted "on the road" at distant national and international locales. This mobile laboratory approach has applications beyond endangered species research, notably for other scientific fields (e.g., studies of hereditary disease in domestic cat models) in which bringing the laboratory to the subject is of value.     

The use of human artificial gametes and the limits of reproductive freedom

Recent developments in generating gametes via in vitro gametogenesis (IVG) from induced pluripotent stem cells (iPSCs) and their successful use for reproductive purposes in animals strongly suggest that soon these methods could also be used in human reproduction. At least two questions emerge in this context: (a) if a legislator should permit their use and (b) if ethical claims emerge that support their provision, e.g., by public health care systems. This urges an ethical reflection of the new reproductive options this technique might offer. Since the concept of reproductive freedom is a key aspect for the ethical evaluation of artificial reproductive technologies (ARTs), it is necessary to analyze if the new possibilities emerging from IVG fall within the scope of this concept. The results may constitute a morally relevant difference between different imaginable applications of IVG and potentially justify differences in claims to access this technology.     

Stereological analysis of nutritive phagocytes and gametogenic cells during the annual reproductive cycle of the green sea urchin, Strongylocentrotus droebachiensis

Abstract. The germinal epithelium of sea urchin gonads contains two interdependent populations of cells: somatic cells called nutritive phagocytes (NPs) and germ cells—ovarian and testicular gonial cells and their derivatives, gametocytes and gametes. Annually, NPs vary their structure and function to produce a changing microenvironment for germ cells during gametogenesis and after spawning. Here, we describe seasonal changes in the NPs as they interact with germ cells during successive gametogenic stages in both sexes of the green sea urchin, Strongylocentrotus droebachiensis. Monthly samples were collected from the Gulf of Maine at a depth of 10 m and analyzed with light microscopy and stereology. Shorter day length and falling seawater temperatures were correlated with nutrient mobilization from NPs, initiation of gonial cell mitosis, and subsequent gametogenesis. During gametogenesis, NPs in both sexes were depleted of nutrients and eventually phagocytize residual ova or spermatozoa. Observations from this study are important for understanding both the cellular aspects of the reproductive biology of sea urchins and those environmental factors that affect the onset and progress of gametogenesis.     

The synergistic activation of FLOWERING LOCUS C by FRIGIDA and a new flowering gene AERIAL ROSETTE 1 underlies a novel morphology in Arabidopsis

The genetic changes underlying the diversification of plant forms represent a key question in understanding plant macroevolution. To understand the mechanisms leading to novel plant morphologies we investigated the Sy-0 ecotype of Arabidopsis that forms an enlarged basal rosette of leaves, develops aerial rosettes in the axils of cauline leaves, and exhibits inflorescence and floral reversion. Here we show that this heterochronic shift in reproductive development of all shoot meristems requires interaction between dominant alleles at AERIAL ROSETTE 1 (ART1), FRIGIDA (FRI), and FLOWERING LOCUS C (FLC) loci. ART1 is a new flowering gene that maps 14 cM proximal to FLC on chromosome V. ART1 activates FLC expression through a novel flowering pathway that is independent of FRI and independent of the autonomous and vernalization pathways. Synergistic activation of the floral repressor FLC by ART1 and FRI is required for delayed onset of reproductive development of all shoot meristems, leading to the Sy-0 phenotype. These results demonstrate that modulation in flowering-time genes is one of the mechanisms leading to morphological novelties.     

Bacterial genomes: evolution of pathogenicity

Bacterial pathogens continue to pose a major threat to economically important plant resources. Disease outbreaks can occur through rapid evolution of a pathogen to overcome host defences. The advent of genome sequencing, especially next-generation technologies, has seen a revolution in the study of plant pathogen evolution over the past five years. This review highlights recent developments in understanding bacterial plant pathogen evolution, enabled by genomics and specifically focusing on type III protein effectors. The genotypic changes and mechanisms involved in pathogen evolution are now much better understood. However, there is still much to be learned about the drivers of pathogen evolution, both in terms of plant resistance and bacterial lifestyle.     

New microinsemination techniques for laboratory animals

Since the development of a reliable mouse intracytoplasmic sperm injection (ICSI) technique in 1995, microinsemination techniques have been widely applied in several laboratory species. As gametes and embryos have specific biological and biochemical features according to the species, technical improvements are necessary for successful microinsemination that subsequently leads to normal fetal development in several species. Recent advanced reproductive research involving genetic engineering often depends on microinsemination techniques that require a high degree of skill, and new human assisted reproductive technology (ART) requires experimental models using laboratory animals. The accumulation of technical improvements in these fields should accelerate the development of microinsemination techniques in mammals, including humans.     

Lineage-Specific Biology Revealed by a Finished Genome Assembly of the Mouse

The mouse (Mus musculus) is the premier animal model for understanding human disease and development. Here we show that a comprehensive understanding of mouse biology is only possible with the availability of a finished, high-quality genome assembly. The finished clone-based assembly of the mouse strain C57BL/6J reported here has over 175,000 fewer gaps and over 139 Mb more of novel sequence, compared with the earlier MGSCv3 draft genome assembly. In a comprehensive analysis of this revised genome sequence, we are now able to define 20,210 protein-coding genes, over a thousand more than predicted in the human genome (19,042 genes). In addition, we identified 439 long, non–protein-coding RNAs with evidence for transcribed orthologs in human. We analyzed the complex and repetitive landscape of 267 Mb of sequence that was missing or misassembled in the previously published assembly, and we provide insights into the reasons for its resistance to sequencing and assembly by whole-genome shotgun approaches. Duplicated regions within newly assembled sequence tend to be of more recent ancestry than duplicates in the published draft, correcting our initial understanding of recent evolution on the mouse lineage. These duplicates appear to be largely composed of sequence regions containing transposable elements and duplicated protein-coding genes; of these, some may be fixed in the mouse population, but at least 40% of segmentally duplicated sequences are copy number variable even among laboratory mouse strains. Mouse lineage-specific regions contain 3,767 genes drawn mainly from rapidly-changing gene families associated with reproductive functions. The finished mouse genome assembly, therefore, greatly improves our understanding of rodent-specific biology and allows the delineation of ancestral biological functions that are shared with human from derived functions that are not.     

PRM谱特征作为分子标签评估精子质量的研究

不孕不育症已成为严重的健康问题,全世界约有15%的育龄夫妇患有这一疾病。辅助生殖技术（ART）和精准医疗是解决这一问题的关键,评估人类精子质量是其中的一个重要环节,但是现行世界卫生组织（WHO）标准诊断手册（第5版）具有不确定性。为了提高评估精子质量的精准性,在本研究中,基于精蛋白（PRM）谱特征和PRM值的评估人类精子质量的新方法,对临床上的病例进行了检测。分析结果表明,运用新方法的精子评估结果与临床结局相一致,优于WHO标准（第5版）的检测结果。这一前瞻性的研究工作对于ART和精准医疗有实际意义：能够在临床应用中提高其成功率和增强其安全性。     

Interactions between the sexes: new perspectives on sexual selection and reproductive isolation

Understanding the processes underlying the origin of new species is a fundamental problem in evolutionary research. Whilst it has long been recognised that closely related taxa often differ markedly in reproductive characteristics, only relatively recently has sexual selection been evoked as a key promoter of speciation through its ability to generate reproductive isolation (RI). Sexual selection potentially can influence the probability that individuals from the same or different populations will reproduce successfully since it shapes precisely those traits involved in mating and reproduction. If reproductive characters diverge along different trajectories, then sexual selection can impact on the evolution of reproductive barriers operating both before and after mating. In this perspective, we consider some new advances in our understanding of the coevolution of male and female sexual signals and receptors and suggest how these developments may provide heretofore neglected insights into the mechanisms by which isolating barriers may emerge. Specifically, we explore how selfish genetic elements (SGEs) can mediate pre- and post-copulatory mate choice, thereby influencing gene flow and ultimately population divergence; we examine evidence from studies of intracellular sperm–egg interactions and propose that intracellular gametic incompatibilities may arise after sperm entry into the egg, and thus contribute to RI; we review findings from genomic studies demonstrating rapid, adaptive evolution of reproductive genes in both sexes and discuss whether such changes are causal in determining RI or simply associated with it; and finally, we consider genetic, developmental and functional mechanisms that might constrain reproductive trait diversification, thereby limiting the scope for reproductive barriers to arise via sexual selection. We hope to stimulate work that will further the understanding of the role sexual selection plays in generating RI and ultimately speciation.     

Food availability and sex reversal in Mytella charruana,an introduced bivalve in the southeastern United States

We studied the reproductive biology of Mytella charruana to determine the potential reproductive success of this newly introduced bivalve species from Central/South America. We analyzed gonad morphology, gametogenesis, and the sex ratios of introduced populations throughout a 12 month period. In the non‐native habitat M. charruana shows the same strategy of gametogenesis that had been observed in its native environment, which is an opportunistic type of gonadal cycle with gametes produced throughout the year. Instead, the spawning period of M. charruana along the southeastern US coast is extended compared to that found in the native environment. We determined the minimum size (shell length) of sexually reproductive mussels to be 1.25 cm. Interestingly, throughout the year the population samples were typically composed of a higher proportion of females. The female to male sex ratio varied within a wide range from 1:0 to 1:3.3. Upon this discovery we tested the effects of food availability on the gametogenesis of adult animals. The sex ratio of mussels collected from different locations and maintained in the laboratory with or without food changed toward a male‐bias under starvation conditions within a month. This is the first study directly showing that food availability can trigger sex reversal in an adult bivalve. According to our data this mussel species will likely continue to spread along the east coast of the US. Moreover, M. charruana may prove to be a model organism in the study of alternative sexuality in bivalves. Mol. Reprod. Dev. 77: 222–230, 2010. © 2009 Wiley‐Liss, Inc.     

Crosstalk between reproductive and immune systems: the teleostean perspective

The bidirectional interaction between the hypothalamic–pituitary-gonadal (HPG) axis and the immune system plays a crucial role in the adaptation of an organism to its environment, its survival and the continuance of a species. Nonetheless, very little is known about this interaction among teleost, the largest group of extant vertebrates. Fishes being seasonal breeders, their immune system is exposed to seasonally changing levels of HPG hormones. On the contrary, the presence and infiltration of leukocytes, the expression of pattern recognition receptors as well as cytokines in gonads suggest their key role in teleostean gametogenesis as in the case of mammals. Moreover, the modulation of gametogenesis and steroidogenesis by lipopolysaccharide implicates the pathological significance of inflammation on reproduction. Thus, it is important to engage in the understanding of the interaction between these two important physiological systems, not only from a phylogenetic perspective but also due to the importance of fish as an important economic resource. In view of this, the authors have reviewed the crosstalk between the reproductive and immune systems in teleosts and tried to explore the importance of this interaction in their survival and reproductive fitness.     

Complex heterochrony underlies the evolution of Caenorhabditis elegans hermaphrodite sex allocation

Hermaphroditic organisms are key models in sex allocation research, yet the developmental processes by which hermaphrodite sex allocation can evolve remain largely unknown. Here we use experimental evolution of hermaphrodite‐male (androdioecious) Caenorhabditis elegans populations to quantify the developmental changes underlying adaptive shifts in hermaphrodite sex allocation. We show that the experimental evolution of increased early‐life self‐fertility occurred through modification of a suite of developmental traits: increased self‐sperm production, accelerated oogenesis and ovulation, and increased embryo retention. The experimental evolution of increased self‐sperm production delayed entry into oogenesis—as expected, given the sequentially coupled production of self‐spermatogenesis and oogenesis. Surprisingly, however, delayed oogenesis onset did not delay reproductive maturity, nor did it trade‐off with gamete or embryo size. Comparing developmental time dynamics of germline and soma indicates that the evolution of increased sperm production did not delay reproductive maturity due to a globally accelerated larval development during the period of self‐spermatogenesis. Overall, heterochrony in gametogenesis and soma can explain adaptive shifts in hermaphrodite sex allocation.     

The malariology centenary

The year 1898 was one of the most significant years in the history of malariology. One hundred years later scientists gathered at the Accademia Nazionale dei Lincei, Rome, to commemorate the Malariology Centenary. This paper provides a short overview of some of the key developments and discoveries in malaria research which took place at the end of the 19th century. The major contributions of Alphonse Laveran, Patrick Manson, Ronald Ross, Battista Grassi and a number of scientists of the Italian School of Malariology to the understanding of the transmission of malaria by Anopheles mosquitoes are described. This paper also highlights the importance of an historical perspective in furthering our understanding of the 'Malaria Challenge after One Hundred Years of Malariology'.     

Preservation of duplicate genes by complementary, degenerative mutations

The origin of organismal complexity is generally thought to be tightly coupled to the evolution of new gene functions arising subsequent to gene duplication. Under the classical model for the evolution of duplicate genes, one member of the duplicated pair usually degenerates within a few million years by accumulating deleterious mutations, while the other duplicate retains the original function. This model further predicts that on rare occasions, one duplicate may acquire a new adaptive function, resulting in the preservation of both members of the pair, one with the new function and the other retaining the old. However, empirical data suggest that a much greater proportion of gene duplicates is preserved than predicted by the classical model. Here we present a new conceptual framework for understanding the evolution of duplicate genes that may help explain this conundrum. Focusing on the regulatory complexity of eukaryotic genes, we show how complementary degenerative mutations in different regulatory elements of duplicated genes can facilitate the preservation of both duplicates, thereby increasing long-term opportunities for the evolution of new gene functions. The duplication-degeneration-complementation (DDC) model predicts that (1) degenerative mutations in regulatory elements can increase rather than reduce the probability of duplicate gene preservation and (2) the usual mechanism of duplicate gene preservation is the partitioning of ancestral functions rather than the evolution of new functions. We present several examples (including analysis of a new engrailed gene in zebrafish) that appear to be consistent with the DDC model, and we suggest several analytical and experimental approaches for determining whether the complementary loss of gene subfunctions or the acquisition of novel functions are likely to be the primary mechanisms for the preservation of gene duplicates. For a newly duplicated paralog, survival depends on the outcome of the race between entropic decay and chance acquisition of an advantageous regulatory mutation.Sidow 1996(p. 717) On one hand, it may fix an advantageous allele giving it a slightly different, and selectable, function from its original copy. This initial fixation provides substantial protection against future fixation of null mutations, allowing additional mutations to accumulate that refine functional differentiation. Alternatively, a duplicate locus can instead first fix a null allele, becoming a pseudogene.Walsh 1995 (p. 426) Duplicated genes persist only if mutations create new and essential protein functions, an event that is predicted to occur rarely.Nadeau and Sankoff 1997 (p. 1259) Thus overall, with complex metazoans, the major mechanism for retention of ancient gene duplicates would appear to have been the acquisition of novel expression sites for developmental genes, with its accompanying opportunity for new gene roles underlying the progressive extension of development itself.Cooke et al. 1997 (p. 362)