Viable Gametes Research Articles

The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis and maintain translational repression of maternal mRNA and oocyte quality.

The regulation of maternal mRNAs is essential for proper oogenesis, the production of viable gametes, and to avoid birth defects and infertility. Many oogenic RNA-binding proteins have been identified with roles in mRNA metabolism, some of which localize to dynamic ribonucleoprotein granules and others that appear dispersed. Here, we use a combination of in vitro condensation assays and the in vivo Caenorhabditis elegans oogenesis model to characterize the properties of the conserved KH-domain MEX-3 protein and to identify novel regulators of MEX-3 and three other translational regulators. We demonstrate that MEX-3 undergoes phase separation and appears to have intrinsic gel-like properties in vitro. We also identify novel roles for the chaperonin-containing tailless complex polypeptide 1 (CCT) chaperonin and actin in preventing ectopic RNA-binding protein condensates in maturing oocytes that appear to be independent of MEX-3 folding. The CCT chaperonin and actin also oppose the expansion of endoplasmic reticulum sheets that may promote ectopic condensation of RNA-binding proteins. These novel regulators of condensation are also required for the translational repression of maternal mRNA which is essential for oocyte quality and fertility. The identification of this regulatory network may also have implications for understanding the role of hMex3 phase transitions in cancer.

Hybridization and growth characteristics of interspecific crosses between Crassostrea hongkongensis and Crassostrea iredalei

Interspecific hybridization of oysters is a highly efficient breeding strategy that can produce commercially viable phenotypes. Crassostrea hongkongensis, the primary cultivar in southern China, faces increasing summer mortality risks and slow growth rates, which elevate commercial risks for farmers. C. iredalei, a commercially cultivated species in Southeast Asia, has been expanding its ecological niche northward in recent years, providing opportunities for interspecific hybridization with C. hongkongensis. This study conducted a 2 × 2 factorial cross between C. hongkongensis and C. iredalei (II: C. iredalei ♀ × C. iredalei ♂, HH: C. hongkongensis ♀ × C. hongkongensis ♂; IH: C. iredalei ♀ × C. hongkongensis ♂, HI: C. hongkongensis ♀ × C. iredalei ♂) and set up 6 salinity treatments (14, 17, 20, 23, 26 and 29 ppt) to explore the optimal salinity for embryo hatching. The results showed that the hybridization was successful, despite bidirectional fertilization and pronounced asymmetry in fertilization success. The optimal hatching salinity for C. iredalei and C. hongkongensis was 23 ppt and 17–20 ppt, respectively, while the optimal salinity for reciprocal hybrids was 20 ppt. Notably, the hatching rate of IH was lower than that of the control group at the optimal salinity. During the pelagic larval stage, hybrid larvae exhibited significant hybrid inferiority, with slower growth rates and higher mortality rates compared to intraspecific hybridization. It is worth noting that IH failed to overcome genetic incompatibility, resulting in complete mortality during the pelagic larval stage and failure to enter the growth stage. At the 360th day of the growth period, HI hybrids grown in Sanbaimen Sea exhibited significant hybrid vigor in shell height (15.65 %), and in Daya Bay Marine Biology Research Station, Chinese Academy of Sciences (MBRS), they showed significant single-parent hybrid advantages in shell height (28.44 %) and cumulative survival rate (58.11 %) compared to C. hongkongensis. In terms of gonadal development, HI hybrids were able to produce functional and viable gametes, although with some delay compared to their parents. The hybrid offspring inherited the phenotypic traits of their parents, with a grayish-yellow flesh color intermediate between the two oyster species. HI is a valuable genetic resource for oyster breeding, and its growth potential should be further explored to meet commercialization demands.

An egg-sabotaging mechanism drives non-Mendelian transmission in mice

Selfish genetic elements drive in meiosis to distort their transmission ratio and increase their representation in gametes, violating Mendel's law of segregation. The two established paradigms for meiotic drive, gamete killing and biased segregation, are fundamentally different. In gamete killing, typically observed with male meiosis, selfish elements sabotage gametes that do not contain them. By contrast, killing is predetermined in female meiosis, and selfish elements bias their segregation to the single surviving gamete (i.e., the egg in animal meiosis). Here, we show that a selfish element on mouse chromosome 2, Responder to drive 2 (R2d2), drives using a hybrid mechanism in female meiosis, incorporating elements of both killing and biased segregation. We propose that if R2d2 is destined for the polar body, it manipulates segregation to sabotage the egg by causing aneuploidy, which is subsequently lethal in the embryo, ensuring that surviving progeny preferentially contain R2d2. In heterozygous females, R2d2 orients randomly on the metaphase spindle but lags during anaphase and preferentially remains in the egg, regardless of its initial orientation. Thus, the egg genotype is either euploid with R2d2 or aneuploid with both homologs of chromosome 2, with only the former generating viable embryos. Consistent with this model, R2d2 heterozygous females produce eggs with increased aneuploidy for chromosome 2, increased embryonic lethality, and increased transmission of R2d2. In contrast to typical gamete killing of sisters produced as daughter cells in a single meiosis, R2d2 prevents production of any viable gametes from meiotic divisions in which it should have been excluded from theegg.

How Per- and Poly-Fluoroalkyl Substances Affect Gamete Viability and Fertilization Capability: Insights from the Literature.

There has been emerging research linking per- and poly-fluoroalkyl substances (PFAS) to gamete viability and fertility. PFAS, prevalent in the environment and water supplies, undergo slow degradation due to their C-F bond and a long half-life (2.3-8.5 years). In females, PFAS inhibit the hypothalamic-pituitary-gonadal (HPG) axis, reducing follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, leading to the inhibition of androgen and estradiol production. PFAS have been found to cause detrimental effects on egg quality through impairing folliculogenesis. In males, PFAS can impair sperm motility and morphology: two fundamental qualities of successful fertilization. PFAS exposure has been proven to inhibit testosterone production, sperm capacitation, and acrosomal reaction. After fertilization, the results of PFAS exposure to embryos have also been investigated, showing reduced development to the blastocyst stage. The aim of this review is to report the main findings in the literature on the impact of PFAS exposure to gamete competency and fertilization capability by highlighting key studies on both male and female fertility. We report that there is significant evidence demonstrating the negative impacts on fertility after PFAS exposure. At high doses, these environmentally abundant and widespread compounds can significantly affect human fertility.

Gamete Viability, Fertilization, and Embryonic Development of Gracious Sea Urchin Tripneustes gratilla Under Varying Temperature Ranges

Gamete Viability, Fertilization, and Embryonic Development of Gracious Sea Urchin Tripneustes gratilla Under Varying Temperature Ranges

Reproductive Conditioning of the Peruvian Scallop Argopecten purpuratus in Different Environments

Obtaining viable Argopecten purpuratus seeds faces challenges, especiallyci the unpredictability of the marine environment and high production costs in hatcheries. However, improving the method of "Broodstock Conditioning In Hatcheries" is key to ensure permanent seed supplies by minimizing the dependence on marine conditions and by maximizing economic viability in hatcheries. In an effort to overcome these barriers, broodstock were conditioned into two different environments: (a) Natural Environment: Natural marine conditions located in Bahía Inglesa, Atacama Region, Chile. (b) Hatchery: Laboratory conditions to achieve gonadal maturation, spawning induction, fertilization and larval development. The purpose of this research was to evaluate how the type of reproductive conditioning affects the reproductive potential and nutritional quality of the progeny. Both methods were successful at inducing the necessary maturity for reproduction, obtaining viable gametes and larvae. On the other hand, it was observed that in the natural environment, the oocytes and D larvae reached a greater size and nutritional value, being the most significant differences with (p < 0.05): the size of the D larvae reached figures of 95.8 ± 3.1 μm and 91.2 ± 2.7 μm in the environment and hatchery, respectively; the lipid content in dry mass was 25.2 ± 3.1 mg g−1 and 13.5 ± 1.9 mg g−1 for the natural environment and hatchery, respectively. Although quality indicators in hatcheries were slightly lower compared to the natural environment, the possibility of conditioning A. purpuratus broodstock independently of environmental variability highlights the importance of further optimizing broodstock conditioning aspects in hatcheries that would allow more predictable and sustainable production.

The Transcriptional Regulator Ume6 is a Major Driver of Early Gene Expression during Gametogenesis.

The process of gametogenesis is orchestrated by a dynamic gene expression program, where a vital subset constitutes the early meiotic genes (EMGs). In budding yeast, the transcription factor Ume6 represses EMG expression during mitotic growth. However, during the transition from mitotic to meiotic cell fate, EMGs are activated in response to the transcriptional regulator Ime1 through its interaction with Ume6. While it is known that binding of Ime1 to Ume6 promotes EMG expression, the mechanism of EMG activation remains elusive. Two competing models have been proposed whereby Ime1 either forms an activator complex with Ume6 or promotes Ume6 degradation. Here, we resolve this controversy. First, we identify the set of genes that are directly regulated by Ume6, including UME6 itself. While Ume6 protein levels increase in response to Ime1, Ume6 degradation occurs much later in meiosis. Importantly, we found that depletion of Ume6 shortly before meiotic entry is detrimental to EMG activation and gamete formation, whereas tethering of Ume6 to a heterologous activation domain is sufficient to trigger EMG expression and produce viable gametes in the absence of Ime1. We conclude that Ime1 and Ume6 form an activator complex. While Ume6 is indispensable for EMG expression, Ime1 primarily serves as a transactivator for Ume6.

All-triploid offspring in the yellowtail tetra Astyanax altiparanae Garutti & Britski 2000 (Teleostei, Characidae) derived from female tetraploid × male diploid crosses.

This study aimed to evaluate the ploidy and survival of larvae resulting from crosses between tetraploid females and diploid males of yellowtail tetra Astyanax altiparanae, both females (three diploids and three tetraploids) and males (n = 3 diploids). Breeders were subjected to hormonal induction with pituitary gland extract from common carp fish (Cyprinus carpio). Females received two doses at concentrations of 0.3 and 3.0 mg/kg -1 body weight and at intervals of 6 h. Males were induced with a single dose of 3.0 mg/kg -1 applied simultaneously with the second dose in females. Oocytes from each diploid and tetraploid female were fertilized with semen from the same male, resulting in two crosses: cross 1 (diploid male and diploid female) and cross 2 (diploid male and tetraploid female). The procedures were performed with separate females (diploid and tetraploid) and diploid males for each repetition (n = 3). For ploidy determination, 60 larvae from each treatment were analyzed using flow cytometry and cytogenetic analyses. As expected, flow cytometry analysis showed that progenies from crosses 1 and 2 presented diploid and triploid individuals, respectively, with a 100% success rate. The same results were confirmed in the cytogenetic analysis, in which the larvae resulting from cross 1 had 50 metaphase chromosomes and those from cross 2 had 75 chromosomes. The oocytes have a slightly ovoid shape at the time of extrusion. Diploid oocytes had a size of 559 ± 20.62 μm and tetraploid of 1025.33 ± 30.91 μm. Statistical differences were observed between eggs from crosses 1 and 2 (P = 0.0130). No significant differences between treatments were observed for survival at the 2-cell stage (P = 0.6174), blastula (P = 0.9717), gastrula (P = 0.5301), somite (P = 0.3811), and hatching (P = 0.0984) stages. In conclusion, our results showed that tetraploid females of the yellowtail tetra A. altiparanae are fertile, present viable gametes after stripping and fertilization using the 'dry method', and may be used for mass production of triploids. This is the first report of these procedures within neotropical characins, and which can be applied in other related species of economic importance.

Sperm Cryopreservation as a Tool for Amphibian Conservation: Production of F2 Generation Offspring from Cryo-Produced F1 Progeny

Sperm cryopreservation and biobanking are emerging as tools for supporting genetic management of small and threatened populations in amphibian conservation programs. However, there is little to no evidence demonstrating reproductive maturity and viability of offspring generated with cryopreserved sperm, potentially limiting widespread integration of these technologies. The purpose of this report is to demonstrate that amphibian sperm can be cryopreserved and thawed to successfully produce individuals of an F1 generation that can reach adulthood and reproductive maturity, to generating viable gametes and an F2 generation. Species-specific exogenous hormones were administered to both F0 and F1 adults to stimulate spermiation and oviposition in the eastern tiger salamander (Ambystoma tigrinum), dusky gopher frog (Lithobates sevosa), and Puerto Rican crested toad (Peltophryne lemur). Sperm cells collected non-lethally from F0 adults were cryopreserved, thawed, and used for in vitro fertilization (IVF) to produce F1 offspring. Individuals of the F1 generation are shown to reach adulthood, express viable gametes, and produce offspring through facilitated breeding, or IVF. The production of amphibian F2 generations shown here demonstrates that amphibian sperm collected non-lethally can be banked and used to generate reproductively viable animals of subsequent generations, thus maintaining valuable genetic linages and diversity in threatened amphibian species. The incredible value that cryopreservation of sperm has for long-term genetic management aids in the sustainability of both in situ and ex situ conservation efforts for this taxon.

Maize cytosolic invertase INVAN6 ensures faithful meiotic progression under heat stress.

Faithful meiotic progression ensures the generation of viable gametes. Studies suggested the male meiosis of plants is sensitive to ambient temperature, but the underlying molecular mechanisms remain elusive. Here, we characterized a maize (Zea mays ssp. mays L.) dominant male sterile mutant Mei025, in which the meiotic process of pollen mother cells (PMCs) was arrested after pachytene. An Asp-to-Asn replacement at position 276 of INVERTASE ALKALINE NEUTRAL 6 (INVAN6), a cytosolic invertase (CIN) that predominantly exists in PMCs and specifically hydrolyses sucrose, was revealed to cause meiotic defects in Mei025. INVAN6 interacts with itself as well as with four other CINs and seven 14-3-3 proteins. Although INVAN6Mei025 , the variant of INVAN6 found in Mei025, lacks hydrolytic activity entirely, its presence is deleterious to male meiosis, possibly in a dominant negative repression manner through interacting with its partner proteins. Notably, heat stress aggravated meiotic defects in invan6 null mutant. Further transcriptome data suggest INVAN6 has a fundamental role for sugar homeostasis and stress tolerance of male meiocytes. In summary, this work uncovered the function of maize CIN in male meiosis and revealed the role of CIN-mediated sugar metabolism and signalling in meiotic progression under heat stress.

The influence of Percoll® density gradient centrifugation before cryopreservation on the quality of frozen wisent (Bison bonasus) epididymal spermatozoa

BackgroundThe wisent (Bison bonasus) is a species that has undergone a population bottleneck. Homozygosity is prevalent within the population and may have a negative impact on semen quality in wisent bulls. Semen samples containing a large amount of functionally and morphologically impaired or dead spermatozoa have lower tolerance for cryopreservation process. Such samples are prone to involve damage acrosomes, to produce and release reactive oxygen which negatively affects proper function of spermatozoas. It is a good practice to select intact and viable gametes before subjecting the sample to cryopreservation to improve the efficiency of this process. The aim of this study was to assess the ability of Percoll® density gradient centrifugation in order to improve the quality of wisent spermatozoa after cryopreservation. Spermatozoa samples were analysed with computer-assisted semen analysis system and flow cytometry.ResultsPercoll® density gradient centrifugation resulted in increased percentage of motile spermatozoa, higher proportion of spermatozoa with normal morphology and proper functionality but also in a significant reduction of the total number of gametes. Nevertheless, the concentration of frozen spermatoza was still sufficient for obtaining a few complete insemination doses suggested for cattle from each epididymis.ConclusionsWhile creating a high-quality genetic reserve, for in vitro fertilisation purposes, eliminating detritus and improving the overall quality of samples is more important than total number of spermatozoa. For these reasons, the achievement of higher post thaw quality of spermatozoa justifies the purification of samples by centrifugation in a Percoll® density gradient prior to the cryopreservation process.

Use of Biological and Synthetic Polymers for Human Spermatozoa Cryopreservation

BACKGROUND: Spermatozoa cryopreservation is an integral part of the assisted reproductive technologies for treatment of infertility. It is also used to preserve the reproductive potential of men. However, using a standard freezing method with glycerol leads to a decrease in morphological and functional characteristics of spermatozoa in the case of oligoasthenoteratozoospermia (OAT). Therefore, it is relevant to develop effective methods of cryopreservation for such sperm. The use of various biopolymers can stabilize the membrane and bind excess water, which forms ice crystals in the medium that causes cell damage when temperature decreases. OBJECTIVE: To study the effectiveness of using cryoprotectant mixtures based on biological and synthetic polymers [serum albumin, polyvinylpyrrolidone (PVP) and insulin] for the cryopreservation of human spermatozoa with OAT. MATERIALS AND METHODS: Human spermatozoa with OAT were cryopreserved using different cryoprotectant media containing 10 % glycerol or 10 % PVP, 20 % albumin and 1 μg/m L human insulin. The viability, motility and mitochondrial membrane potential of spermatozoa were assessed after rewarming. RESULTS: A cryoprotectant solution containing 10 % PVP, 20 % human serum albumin and 1 μg/m L insulin enabled a similar level (%) of viable gametes compared with the standard method using glycerol, while the number of motile cells was significantly lower (p < 0.008). The membrane mitochondrial potential did not differ significantly from fresh sperm. CONCLUSION: The data obtained in this study show the effectiveness of a biopolymer mixture containing PVP, serum albumin and insulin for the cryopreservation of human OAT spermatozoa.

P-117 A novel method to select spermatozoa with the best morphokinetic characteristics and superior genomic integrity

Abstract Study question Can an automated device based on membrane electrophoresis be used to select spermatozoa with superior morphokinetic characteristics and lower sperm chromatin fragmentation (SCF)? Summary answer Albeit at an expense of concentration, a membrane-electrophoretic device was able to isolate a proportion of spermatozoa with the highest motility, morphology, and genomic integrity. What is known already Conventional sperm selection based on density gradient centrifugation (DGC) can enhance the proportion of progressively motile spermatozoa; however, DGC methods are limited in their ability to select spermatozoa with higher genomic integrity. Membrane-based microfluidic technologies (MFSS) have already been used clinically to select for spermatozoa with a superior chromatin status; however, these methods are based on the intrinsic characteristics of motile spermatozoa capable of selecting themselves. By incorporating an extrinsic electrophoretic drive, membrane electrophoresis would overcome a limitation of progressive motility in the semen sample and be able to isolate viable gametes with better morphology and reduced SCF. Study design, size, duration From August 2020 to December 2021, semen samples from 68 men were evaluated by standard semen analysis and simultaneously processed by DGC or a novel membrane-electrophoretic sperm sorter (EPSS) to select for progressively motile spermatozoa. Concentration, motility, progressive motility, morphology, and SCF were measured and compared between raw, DGC-, and EPSS-processed specimens. Participants/materials, setting, methods Fresh ejaculates were evaluated by standard semen analysis according to WHO 2021 criteria. Following complete liquefaction, specimens were divided into two equal aliquots for DGC and EPSS sorting. SCF was assessed by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay under fluorescent microscopy; at least 500 spermatozoa were evaluated for each specimen with a normal threshold of ≤ 15%. Main results and the role of chance A total of 68 men with an average age of 38.1±6 years had the following average semen parameters: volume of 3.1±1 mL, concentration of 58.2±33 x106/mL, 43.3±9% motility, 40.5±9% forward progression motility, and a normal morphology of 2.9±0.8%. When comparing the final parameters between the EPSS and DGC methods, despite a reduced sperm concentration (7.3±9 x106/mL vs 45.0±36 x106/mL, P<0.0001, respectively), EPSS was highly selective for motile spermatozoa, yielding significantly higher motility (93.1±16% vs. 86.8±15% P<0.0001) and forward progressive motility (92.3±17% vs 85.2±15%, P<0.0001). Despite an overall lower sperm recovery rate (21.4±25% vs 75.4±17%, P<0.0001), normal morphology improved to 3.4±0.8% (P<0.0001) after EPSS but remained unchanged in the DGC-processed sample. Although both EPSS and DGC improved SCF from 12.2±9% in the raw specimen to 4.8±6% and 6.6±7% (P < 0.0001), respectively, EPSS outperformed DGC (P < 0.05). Moreover, the DGC method took up to 60 min to process, whereas the EPSS technique took a total of 6 min. Limitations, reasons for caution The selection of spermatozoa by EPSS is a promising technique to isolate progressively motile spermatozoa with enhanced morphology and superior chromatin integrity, albeit at a lower concentration. This is a preliminary study; the benefit and safety of this method must be further demonstrated by insemination or IVF treatment. Wider implications of the findings A membrane electrophoresis device may be a viable alternative method to MFSS to identify spermatozoa with superior morphokinetic characteristics and intact chromatin. Moreover, incorporating an automated device can reduce gamete processing time while minimizing labor costs and inter-operator errors. Trial registration number N/A

Gametogenesis in Plasmodium: Delving Deeper to Connect the Dots.

In the coming decades, eliminating malaria is the foremost goal of many tropical countries. Transmission control, along with an accurate and timely diagnosis of malaria, effective treatment and prevention are the different aspects that need to be met synchronously to accomplish the goal. The current review is focused on one of these aspects i.e., transmission control, by looking deeper into the event called gametogenesis. In the Plasmodium life cycle, gametocytes are the first life forms of the sexual phase. The transmission of the parasite and the disease is critically dependent on the number, viability and sex ratio of mature gametocytes and their further development inside mosquito vectors. Gametogenesis, the process of conversion of gametocytes into viable gametes, takes place inside the mosquito midgut, and is a tightly regulated event with fast and multiple rounds of DNA replication and diverse cellular changes going on within a short period. Interrupting the gametocyte-gamete transition is ought to restrict the successful transmission and progression of the disease and hence an area worth exploring for designing transmission-blocking strategies. This review summarizes an in-depth and up-to-date understanding of the biochemical and physiological mechanism of gametogenesis in Plasmodium, which could be targeted to control parasite and malaria transmission. This review also raises certain key questions regarding gametogenesis biology in Plasmodium and brings out gaps that still accompany in understanding the spectacular process of gametogenesis.

Capture of Spawning Brook Trout by Electrofishing Does Not Impair Embryo Survival

AbstractElectrofishing is widely used to capture fish in freshwater systems. Fisheries assessment and fish culture activities that occur during spawning do so in the absence of a meaningful base of evidence about the potential effects of electrofishing on fish reproduction. In this laboratory experiment, we assessed whether electrofishing adult Brook Trout Salvelinus fontinalis affected the survival of their embryos. We used two genetically distinct strains of Brook Trout (domestic and wild‐origin strains) to do so. Both strains and sexes of fish were exposed to pulsed‐DC electrofishing techniques in a fully factorial design (i.e., male shocked, female shocked, both parents shocked, or neither parent shocked [control]), after which their incubating offspring were monitored for survival to the eyed egg, alevin, and fry stages. We did not detect any effects of our electrofishing treatment or interactions with the sex or strain of the fish exposed to electrofishing, suggesting that electrofishing did not negatively impact gamete viability. Our results support the use of responsible electrofishing to collect spawning salmonids for the purpose of gamete collection for hatchery rearing.

The Arabidopsis HOP2 gene has a role in preventing illegitimate connections between nonhomologous chromosome regions.

Meiotic homologous chromosomes synapse and undergo crossing over (CO). In many eukaryotes, both synapsis and crossing over require the induction of double stranded breaks (DSBs) and subsequent repair via homologous recombination. In these organisms, two key proteins are recombinases RAD51 and DMC1. Recombinase-modulators HOP2 and MND1 assist RAD51 and DMC1 and also are required for synapsis and CO. We have investigated the hop2-1 phenotype in Arabidopsis during the segregation stages of both meiosis and mitosis. Despite a general lack of synapsis during prophase I, we observed extensive, stable interconnections between nonhomologous chromosomes in diploid hop2-1 nuclei in first and second meiotic divisions. Using γH2Ax as a marker of unrepaired DSBs, we detected γH2AX foci from leptotene through early pachytene but saw no foci from mid-pachytene onward. We conclude that the bridges seen from metaphase I onward are due to mis-repaired DSBs, not unrepaired ones. Examining haploids, we found that wild type haploids produce only univalents, but hop2-1 haploids like hop2-1 diploids have illegitimate connections stable enough to produce bridged chromosomes during segregation. Our results suggest that HOP2 has a significant active role in preventing repairs that use nonhomologous chromosomes during meiosis. We also found evidence that HOP2 plays a role in preventing illegitimate repair of radiation-induced DSBs in rapidly dividing petal cells. We conclude that HOP2 in Arabidopsis plays both a positive role in promoting synapsis and a separable role in preventing DSB repair using nonhomologous chromosomes. SIGNIFICANCE STATEMENT : The fidelity of homologous recombination (HR) during meiosis is essential to the production of viable gametes and for maintaining genome integrity in vegetative cells. HOP2 is an important protein for accurate meiotic HR in plants. We have found evidence of high levels of illegitimate repairs between nonhomologous chromosomes during meiosis and in irradiated petal cells in hop2-1 mutants, suggesting a role for HOP2 beyond its established role in synapsis and crossing over.

Arsenic exposure and its implications in male fertility.

Arsenic exposure is a global health concern. This toxic metalloid is ubiquitous in the environment and contaminates food and drinking water. Once ingested, it undergoes a complex metabolic process within the body, which contributes to its accumulation and reactivity. Arsenic toxicity stems from the induction of oxidative stress, inhibition of thiol-containing proteins, and mimicry of inorganic phosphates. Arsenic poisoning is associated with the development of reproductive disorders. In males, arsenic causes a reduction in testicular weight and alterations in steroidogenesis and spermatogenesis. Moreover, it reduces the number and quality of spermatozoa harvested from the cauda epididymis. The mitochondria are targets of arsenic toxicity because of the production of free radicals and their high content of cysteine-rich proteins and fatty acids. Mitochondrial dysfunction may contribute to reproductive disorders because this organelle is crucial for controlling testicular and epididymal events related to sperm production and maturation. All of these alterations mediated by arsenic exposure contribute to the failure of male reproductive competence by reducing gamete viability. This review describes the potential mechanisms of arsenic toxicity, its detrimental effects on male reproductive organs, and consequences on sperm fertility.

Translating Basic Research to Animal Agriculture.

Procedures to maintain viability of mammalian gametes and embryos in vitro, including cryopreservation, have been exceedingly valuable for my research over the past 55 years. Keeping sperm viable in vitro enables artificial insemination, which, when combined with selective breeding, often is the most effective approach to making rapid genetic change in a population. Superovulation and embryo transfer constitute a parallel approach for amplifying reproduction of female mammals. More recent developments include sexing of semen, in vitro fertilization, cloning by nuclear transfer, and genetic modification of germline cells, tools that are enabled by artificial insemination and/or embryo transfer for implementation. I have been fortunate in being able to contribute to the development of many of the above techniques, and to use them for research and applications for improving animal agriculture. Others have built on this work to circumvent human infertility, assist reproduction of companion animals, and rescue endangered species. It also has been a privilege to teach, mentor, and be mentored in this area. Resulting worldwide friendships have enriched me personally and professionally.

Transcription-wide mapping of dihydrouridine reveals that mRNA dihydrouridylation is required for meiotic chromosome segregation

The epitranscriptome has emerged as a new fundamental layer of control of gene expression. Nevertheless, the determination of the transcriptome-wide occupancy and function of RNA modifications remains challenging. Here we have developed Rho-seq, an integrated pipeline detecting a range of modifications through differential modification-dependent rhodamine labeling. Using Rho-seq, we confirm that the reduction of uridine to dihydrouridine (D) by the Dus reductase enzymes targets tRNAs in E.coli and fission yeast. We find that the D modification is also present on fission yeast mRNAs, particularly those encoding cytoskeleton-related proteins, which is supported by large-scale proteome analyses and ribosome profiling. We show that the α-tubulin encoding mRNA nda2 undergoes Dus3-dependent dihydrouridylation, which affects its translation. The absence of the modification on nda2 mRNA strongly impacts meiotic chromosome segregation, resulting in low gamete viability. Applying Rho-seq to human cells revealed that tubulin mRNA dihydrouridylation is evolutionarily conserved.

Shallow-emerged coral may warn of deep-sea coral response to thermal stress

In the Gulf of Alaska, commercially harvested fish species utilize habitats dominated by red tree corals (Primnoa pacifica) for shelter, feeding, and nurseries, but recent studies hint that environmental conditions may be interrupting the reproductive lifecycle of the corals. The North Pacific has experienced persistent and extreme thermal variability in recent years and this pattern is predicted to continue in coming decades. Recent discovery of deep-water emerged coral populations in Southeast Alaska fjords provided opportunity for detailed life-history studies and comparison to corals in managed habitats on the continental shelf. Here we show that sperm from deep colonies develops completely, but in shallow colonies, sperm development is prematurely halted, likely preventing successful production of larvae. We hypothesize that the divergence is due to differing temperature regimes presently experienced by the corals. Compared to deep populations below the thermocline, shallow populations experience much greater seasonal thermal variability and annual pulses of suspected near-lethal temperatures that appear to interrupt the production of viable gametes. The unique opportunity to comprehensively study emerged populations presently affected by thermal stress provides advance warning of the possible fate of deep corals in the Gulf of Alaska that will soon experience similar ocean conditions.