Paternal Contribution Research Articles

Altered Developmental Programming in a Dual Model of Maternal and Paternal Alcohol Consumption

According to the CDC, Fetal Alcohol Spectrum Disorders (FASDs) are a group of conditions that can occur in a person whose mother drank alcohol during pregnancy. This definition places the fault for FASD-associated mental and physical defects exclusively on the mother. However, emerging research can now link preconception paternal environmental exposures and lifestyle choices with several disease states within the offspring. Despite the efforts to recognize paternal epigenetic contributions, research examining congenital disabilities is limited and remains exclusively focused on maternal exposures. We hypothesized that FASD-associated developmental defects would become exacerbated in a dual parental model of alcohol consumption. To investigate the contributions of dual parental ethanol (EtOH) exposure on offspring development, we exposed C57BL/6J adult [postnatal day (PND) 90] males and females to either control or alcohol (10% EtOH) treatments using the Drinking in the Dark (DID) method. After a preconception treatment period of 6 weeks for males and ten days for females, we mated exposed mice. Pregnancies were confirmed by weight gain at gestational day (GD) 10. Pregnant females ceased alcohol treatments at gestational day ten, and we terminated pregnancies at either GD 16.5 or 18.5. We examined morphometric measures of fetal development and 2D images of the placenta and craniofacial features. From these studies, we find that each exposure source (maternal, paternal, or dual parental) exhibits its unique growth and craniofacial phenotype. This finding draws attention to the need for a thorough investigation of both parents' exposure history when determining the child's overall health status. It also emphasizes that the presence of a defect does not mean that the blame lies solely with the mother. Instead, it might be the combinatorial effect of both parents' life exposures before conception.

Prenatal Exposure to Persistent Organic Pollutants and Maternal Folic Acid Supplementation: Their Impact on Glucose Homeostasis in Male Rat Descendants

Exposure to persistent organic pollutants (POPs) is associated with insulin resistance while folic acid (FA) may offer a protective effect. However, the paternal contribution to metabolic phenotypes in offspring is not well known yet. Hence, we investigated whether maternal exposure to POPs affects glucose homeostasis and whether maternal FA supplementation counteracts POP effects transmitted via male descendants. Sprague–Dawley founder dams (F0) were fed a diet containing 2 or 6 mg/kg of FA and were force-fed with either a POP mixture or corn oil for 9 weeks. Subsequent male descendants did not receive any treatment. Blood glucose, plasma insulin and C-peptide were measured during an oral glucose tolerance test in males aged 90 and 180 days from generation 1 (F1), 2 (F2) and 3 (F3). Prenatal POP exposure increased fasting glucose in 90-day-old F1 males and C-peptide in 90-day-old F2 males. Prenatal FA supplementation decreased C-peptide in 90 and 180-day-old F1 males. In 180-day-old F3 males, FA supplementation counteracted POPs on fasting and postglucose C-peptide, indicating reduced insulin secretion. Prenatal exposure to an environmentally relevant POP mixture caused abnormalities in glucose homeostasis that are transmitted from one generation to the next through the paternal lineage. Prenatal FA supplementation counteracted some of the deleterious effects of POPs on glucose homeostasis.

Parental genetic contributions to neonatal temperament in a nonhuman primate (Macaca mulatta) model.

Temperament is an individual's nature and is widely believed to have a heritable foundation. Few studies, however, have evaluated paternal and maternal contributions to the triadic dimensions of temperament. Rhesus monkeys are widely utilized to model genetic contributions to human development due to their close genetic-relatedness and common temperament structure, providing a powerful translational model for investigating paternal and maternal genetic influences on temperament. The temperament of rhesus monkey infants born to 19 different sires and 50 different dams was assessed during the first month of life by comparing the temperament of paternal or maternal half-siblings reared with their mothers in species-normative conditions or reared in a neonatal nursery. Factor scores from three dimensions of temperament were obtained (Orienting/Regulation, Negative Affectivity, and Surgency/Extraversion) and ANOVAs were used to assess genetic effects. For paternal half-siblings, results showed a statistically significant paternal contribution to Orienting/Regulation, Negative Affectivity, and Surgency/Extraversion factor scores. For maternal half-siblings, results showed a statistically significant contribution to Orienting/Regulation factor scores. When parsed by early rearing condition, results showed a paternal contribution Orienting/Regulation, Negative Affectivity, and Surgency/Extraversion scores for paternal half-siblings reared in the neonatal nursery, while there was only a paternal contribution to Surgency/Extraversion for paternal half-siblings reared by their mothers. There was only a maternal contribution to Orienting/Regulation for maternal half-siblings reared by their mothers. These results show that paternal and maternal contributions to temperament vary by environmental context, and that mothers may environmentally buffer their infants from paternal contributions to their temperament.

Altered mitochondrial function in spermatozoa from patients with repetitive fertilization failure after ICSI revealed by proteomics.

Unexplained fertilization failure (FF), occurring in 1-3% of intracytoplasmic sperm injection (ICSI) cycles, results in both psychological and financial burden for the patients. However, the molecular causes behind FF remain largely unknown. Mass spectrometry is a powerful technique to identify and quantify proteins across samples; however, no study so far has used it to dissect the proteomic signature of spermatozoa with FF after ICSI. To investigate whether sperm samples from patients suffering repetitive FF after ICSI display alterations in their protein content. Seventeen infertile men were included: 5 patients presented FF in ≥3 consecutive ICSI cycles, while 12 patients had a fertilization rate >75% (controls). Individual sperm samples were subjected to 2D-LC-MS/MS. Both conventional and novel statistical approaches were used to identify differentially abundant proteins. Additionally, analysis of mitochondrial and proteasomal abundance and activity were performed, using Western blot, FACS analysis of JC-1 staining and AMC-peptide fluorometric assay. Four proteins presented lower abundance (FMR1NB, FAM209B, RAB2B, and PSMA1) in the FF group compared to controls, while five mitochondrial proteins presented higher abundance in FF (DLAT, ATP5H, SLC25A3, SLC25A6, and FH) (p<0.05). The altered abundance of mitochondrial DLAT and proteasomal PSMA1 was corroborated by Western blot. Of relevance, novel stable-protein pair analysis identified 73 correlations comprising 28 proteins within controls, while different mitochondrial proteins (ie, PDHA2, PHB2, and ATP5F1D) lost >50% of these correlations in specific FF samples pointing out specific mitochondrial deregulations. This is the first proteomic analysis of spermatozoa from patients who resulted in fertilization failure after ICSI. The altered proteins, most of them related to mitochondrial function, could help to identify diagnostic/prognostic markers of fertilization failure and could further dissect the molecular paternal contribution to reach successful fertilization. Sperm samples from patients with FF after ICSI present altered abundance of different proteins, including mainly mitochondrial proteins.

Males That Silence Their Father's Genes: Genomic Imprinting of a Complete Haploid Genome.

Genetic conflict is considered a key driver in the evolution of reproductive systems with non-Mendelian inheritance, where parents do not contribute equally to the genetic makeup of their offspring. One of the most extraordinary examples of non-Mendelian inheritance is paternal genome elimination (PGE), a form of haplodiploidy which has evolved repeatedly across arthropods. Under PGE, males are diploid but only transmit maternally inherited chromosomes, while the paternally inherited homologues are excluded from sperm. This asymmetric inheritance is thought to have evolved through an evolutionary arms race between the paternal and maternal genomes over transmission to future generations. In several PGE clades, such as the mealybugs (Hemiptera: Pseudococcidae), paternal chromosomes are not only eliminated from sperm, but also heterochromatinized early in development and thought to remain inactive, which could result from genetic conflict between parental genomes. Here, we present a parent-of-origin allele-specific transcriptome analysis in male mealybugs showing that expression is globally biased toward the maternal genome. However, up to 70% of somatically expressed genes are to some degree paternally expressed, while paternal genome expression is much more restricted in the male reproductive tract, with only 20% of genes showing paternal contribution. We also show that parent-of-origin-specific gene expression patterns are remarkably similar across genotypes, and that genes with completely biparental expression show elevated rates of molecular evolution. Our results provide the clearest example yet of genome-wide genomic imprinting in insects and enhance our understanding of PGE, which will aid future empirical tests of evolutionary theory regarding the origin of this unusual reproductive strategy.

Centriolar defects, centrin 1 alterations, and FISH studies in human spermatozoa of a male partner of a couple that produces aneuploid embryos in natural and artificial fertilization

PurposeTo study the potential paternal contribution to aneuploidies in the man of a couple who obtained trisomic embryos with natural and assisted fertilization.MethodsSemen analysis, immunofluorescence for localization of tubulin and centrin 1, transmission electron microscopy (TEM), and fluorescence in situ hybridization (FISH) analysis for chromosomes 18 and 9 were performed. Sperm of fertile men were used as controls.ResultsThe percentages of sperm motility and normal forms were decreased. The percentages of sperm with tail reduced in dimension, headless tails, coiled tails, and altered head-tail junction were significantly higher (P < 0.01) in the patient than in controls, whereas the percentage of sperm with a normal centrin 1 localization (two spots in the centriolar area) was significantly reduced (P < 0.01) in the patient. Immunofluorescence with anti-tubulin antibody showed that in most of the patient’s sperm connecting pieces (83.00 ± 1.78%), two spots were present, indicating prominent proximal centriole/centriolar adjunct and evident distal centriole, whereas controls’ sperm displayed a single spot, indicating the proximal centriole. The percentage of sperm with two spots was significantly higher (P < 0.01) in the patient than in controls. TEM analysis showed that centriolar adjuncts of the patient’s sperm were significantly longer (721.80 ± 122.26 nm) than in controls’ sperm (310.00 ± 64.11 nm; P < 0.001). The aneuploidy frequencies of the patient’s sperm, detected by FISH analysis, were increased with respect to controls.ConclusionA paternal contribution to sperm aneuploidies cannot be excluded since the patient’s sperm showed altered morphology, immature centriolar adjunct, presence of evident distal centriole, scarce presence of centrin 1, and high aneuploidy frequency.

Comparative Embryogenesis in Angiosperms: Activation and Patterning of Embryonic Cell Lineages.

Following fertilization in flowering plants (angiosperms), egg and sperm cells unite to form the zygote, which generates an entire new organism through a process called embryogenesis. In this review, we provide a comparative perspective on early zygotic embryogenesis in flowering plants by using the Poaceae maize and rice as monocot grass and crop models as well as Arabidopsis as a eudicot model of the Brassicaceae family. Beginning with the activation of the egg cell, we summarize and discuss the process of maternal-to-zygotic transition in plants, also taking recent work on parthenogenesis and haploid induction into consideration. Aspects like imprinting, which is mainly associated with endosperm development and somatic embryogenesis, are not considered. Controversial findings about the timing of zygotic genome activation as well as maternal versus paternal contribution to zygote and early embryo development are highlighted. The establishment of zygotic polarity, asymmetric division, and apical and basal cell lineages represents another chapter in which we also examine and compare the role of major signaling pathways, cell fate genes, and hormones in early embryogenesis. Except for the model Arabidopsis, little is known about embryopatterning and the establishment of the basic body plan in angiosperms. Using available in situ hybridization, RNA-sequencing, and marker data, we try to compare how and when stem cell niches are established. Finally, evolutionary aspects of plant embryo development are discussed.

Fixation of allelic gene expression landscapes and expression bias pattern shape the transcriptome of the clonal Amazon molly

The Amazon molly is a unique clonal fish species that originated from an interspecies hybrid between Poecilia species P. mexicana and P. latipinna. It reproduces by gynogenesis, which eliminates paternal genomic contribution to offspring. An earlier study showed that Amazon molly shows biallelic expression for a large portion of the genome, leading to two main questions: (1) Are the allelic expression patterns from the initial hybridization event stabilized or changed during establishment of the asexual species and its further evolution? (2) Is allelic expression biased toward one parental allele a stochastic or adaptive process? To answer these questions, the allelic expression of P. formosa siblings was assessed to investigate intra- and inter-cohort allelic expression variability. For comparison, interspecies hybrids between P. mexicana and P. latipinna were produced in the laboratory to represent the P. formosa ancestor. We have identified inter-cohort and intra-cohort variation in parental allelic expression. The existence of inter-cohort divergence suggests functional P. formosa allelic expression patterns do not simply reflect the atavistic situation of the first interspecies hybrid but potentially result from long-term selection of transcriptional fitness. In addition, clonal fish show a transcriptional trend representing minimal intra-clonal variability in allelic expression patterns compared to the corresponding hybrids. The intra-clonal similarity in gene expression translates to sophisticated genetic functional regulation at the individuum level. These findings suggest the parental alleles inherited by P. formosa form tightly regulated genetic networks that lead to a stable transcriptomic landscape within clonal individuals.

62 Investigating differences in gene expression between invitro-produced bovine embryos and parthenotes

Parthenogenic activation allows for the development of an oocyte without male gamete contribution and may serve as a suitable model for understanding maternal and paternal contributions during development. In the bovine, parthenotes lack the ability to survive to term once transferred into females. The goal of this study was to investigate the gene expression profile at the blastocyst stage in parthenotes and embryos by characterising expression of developmentally important genes, such as markers for pluripotency (OCT4, NANOG), hypoblast (GATA6), epiblast (SOX2), trophectoderm (CDX2), and maternal-embryo communication (IFNT2). To test this, IVM oocytes were either fertilized to a bull with known fertility invitro or activated. To activate, oocytes were denuded, and placed in ionomycin calcium salt for 5min. They were incubated for 3h in 6-(dimethylamino)purine (6-DMAP) and placed in synthetic oviductal fluid (SOF). Putative zygotes and activated oocytes were cultured in SOF for 8 days. Cleavage (at least one cellular division) was recorded on Day 3 and development to the blastocyst stage was recorded on Day 8 after insemination or activation. Cleavage rate was 85.9% and 83.2% for parthenotes and embryos, respectively. Both groups produced a similar blastocyst rate, 32.3% for parthenotes and 33.7% for embryos. On Day 8, blastocyst stage parthenotes and embryos were collected. Data were analysed by ANOVA (Tukey HSD post hoc test) using the GLM procedure of SAS version 9.4. Pools of 5 embryos or parthenotes (3 replicates) were flash frozen and stored until RNA isolation (PicoPure™ RNA Isolation Kit). Real-time PCR was used for quantification of gene expression. Genes were analysed relative to a housekeeping gene, GAPDH. There was no difference in gene expression detected for OCT4 (P=0.25), NANOG (P=0.11), GATA6 (P=0.32), SOX2 (P=0.25), or IFNT2 (P=0.52). Expression of CDX2 was lower in the parthenotes than the embryos (P=0.05). In a second experiment, the proteins GATA6, NANOG, and CDX2 were immunolocalized in 17 parthenotes and 15 embryos. Fixed embryos were permeabilized, blocked, and placed in primary antibodies overnight. After, they were placed into the secondary antibody for 1h, followed by nuclear stain. There was a decreased mean intensity of CDX2 in the parthenotes compared to embryos (P=0.005). No difference (P&amp;gt;0.05) in GATA6 or NANOG was observed between the 2 groups. The ratio of inner cell mass to trophectoderm was higher (P=0.04) in the parthenotes (2.5±0.23) than in the embryos (1.7±0.25). Here we analysed and confirmed the expression of developmentally important genes at the blastocyst stage in embryos and parthenotes. CDX2, a marker of the trophectoderm that will later give rise to the placenta, was downregulated in parthenotes. This highlights the importance of the contribution of the paternal genome to development. Further research is necessary to elucidate the ability of the parthenotes to establish and maintain pregnancy. This research was supported by the National Needs Fellowship funded by USDA NIFA Grant 2019-38420-28972.

Within-season shifts in multiple paternity patterns in mass-nesting olive ridley sea turtles

Multiple paternity is common to all sea turtle species, but its causes and consequences are hard to ascertain and the behaviors and success of males difficult to observe. This study aims to describe patterns of multiple paternity for olive ridley turtles Lepidochelys olivacea at Playa de Escobilla, an ‘arribada’ (mass-nesting) site on the Mexican Pacific coast with over a million clutches laid each reproductive season. A total of 15 females and their hatchlings were sampled during 3 arribada events which occurred over the 2016-2017 nesting season. Females and hatchlings (N = 329) were genotyped at 5 microsatellite loci, from which we inferred the alleles of 46 contributing males. Multiple paternity was detected in 60% of the analyzed clutches, which were sired by a range of 2 to 7 males. Multiple paternity rates differed significantly across arribada events, suggesting more males achieved fertilizations earlier in the breeding season. Paternal contribution in 6 of the clutches with multiple paternity was skewed towards a single male; the remaining clutches had a homogeneous male contribution. However, our results are based on relatively small within-arribada sample sizes. The frequency of multiple paternity among turtle clutches laid on this arribada beach could be related to the density of breeding individuals in the reproductive patch off Playa de Escobilla, rather than to the nesting population size or female size.

Anything New about Paternal Contribution to Reproductive Outcomes? A Review of the Evidence.

Paternal health and behavioral lifestyles affect reproductive and neonatal outcomes and yet the magnitude of these effects remain underestimated. Even though these impacts have been formally recognized as a central aspect of reproductive health, health care services in Europe often neglect the involvement of fathers in their reproductive programs. Following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines for systematic reviews, a literature search was carried out to assess the possible impact of paternal health on reproductive outcomes. The comprehensive strategy included cohort studies and meta-analysis available on PubMed, Web of Science, CINAHL, and Google scholar. Cross-referencing of bibliographies of the selected papers ensured wider study capture. Paternal factors were grouped into two categories respectively identified with the terms “Biological Paternal Factors” and “Lifestyle Paternal Factors”. Advanced age may impair male fertility and affect early pregnancy stages. Increased body mass index, smoking, alcohol and recreational drugs, all alter seminal fluid parameters. Hazardous alcohol use correlates with low birthweight in pregnancy and harmful behavioral lifestyles have been linked to congenital heart defects, metabolic and neurodevelopmental disorders in the offspring. Measures targeting paternal health and lifestyle within the first 1,000 days' timeframe need to be implemented in couples undergoing reproductive decisions. Health professionals, as well as future fathers, must be aware of the benefits for the offspring associated with correct paternal behaviors. More research is needed to build guidelines and to implement specific programs aiming at reproductive health promotion.

Paternal contributors in recurrent pregnancy loss: Cues from comparative proteome profiling of seminal extracellular vesicles.

Recent evidence entail paternal factors as plausible contributors in spontaneous recurrent pregnancy loss (RPL). Seminal extracellular vesicles secreted from cells of male reproductive tract carry regulatory proteins and RNAs. They are proposed to regulate sperm maturation and function while their fusion to endometrial stromal cells helps in decidualization. Nevertheless, the mechanism(s) involved in these processes are poorly understood. This study aims at elucidating the molecular basis of paternal contribution by comparative proteomics (label-free LC-MS/MS) of isolated seminal extracellular vesicles from fertile men and partners of patients with RPL (n = 21 per group). Bioinformatics analysis revealed the identified differentially expressed proteins to be involved in DNA replication, recombination and repair, gene expression, cellular assembly and organization, cell death, and survival. Major disease pathways affected were identified as developmental, hereditary, and immunological disorders. Of the three identified hub genes regulating the above disease pathways, two (HNRNPC and HNRNPU) are overexpressed while RUVBL1 is underexpressed along with over expression of HIST1H1C, DDX1, surmising defective chromatin packaging, and histone removal in spermatozoa resulting in improper expression in paternal genes thereby leading to abnormal embryo development. Besides, alteration in GSTP1 expression points oxidative predominance in RPL group. Differential expression of C3, C4a/C4b, CFB, and GDF 15 may be involved in altered maternal immune response to paternal antigens resulting in impaired decidualization.

Germline transmission of donor, maternal and paternal mtDNA in primates.

What are the long-term developmental, reproductive and genetic consequences of mitochondrial replacement therapy (MRT) in primates? Longitudinal investigation of MRT rhesus macaques (Macaca mulatta) generated with donor mtDNA that is exceedingly distant from the original maternal counterpart suggest that their growth, general health and fertility is unremarkable and similar to controls. Mitochondrial gene mutations contribute to a diverse range of incurable human disorders. MRT via spindle transfer in oocytes was developed and proposed to prevent transmission of pathogenic mtDNA mutations from mothers to children. The study provides longitudinal studies on general health, fertility as well as transmission and segregation of parental mtDNA haplotypes to various tissues and organs in five adult MRT rhesus macaques and their offspring. MRT was achieved by spindle transfer between metaphase II oocytes from genetically divergent rhesus macaque populations. After fertilization of oocytes with sperm, heteroplasmic zygotes contained an unequal mixture of three parental genomes, i.e. donor (≥97%), maternal (≤3%), and paternal (≤0.1%) mitochondrial (mt)DNA. MRT monkeys were grown to adulthood and their development and general health was regularly monitored. Reproductive fitness of male and female MRT macaques was evaluated by time-mated breeding and production of live offspring. The relative contribution of donor, maternal, and paternal mtDNA was measured by whole mitochondrial genome sequencing in all organs and tissues of MRT animals and their offspring. Both male and female MRT rhesus macaques containing unequal mixture of three parental genomes, i.e. donor (≥97%), maternal (≤3%), and paternal (≤0.1%) mtDNA reached healthy adulthood, were fertile and most animals stably maintained the initial ratio of parental mtDNA heteroplasmy and donor mtDNA was transmitted from females to offspring. However, in one monkey out of four analyzed, initially negligible maternal mtDNA heteroplasmy levels increased substantially up to 17% in selected internal tissues and organs. In addition, two monkeys showed paternal mtDNA contribution up to 33% in selected internal tissues and organs. N/A. Conclusions in this study were made on a relatively low number of MRT monkeys, and on only one F1 (first generation) female. In addition, monkey MRT involved two wildtype mtDNA haplotypes, but not disease-relevant variants. Clinical trials on children born after MRT will be required to fully determine safety and efficacy of MRT for humans. Our data show that MRT is compatible with normal postnatal development including overall health and reproductive fitness in nonhuman primates without any detected adverse effects. 'Mismatched' donor mtDNA in MRT animals even from the genetically distant mtDNA haplotypes did not cause secondary mitochondrial dysfunction. However, carry-over maternal or paternal mtDNA contributions increased substantially in selected internal tissues / organs of some MRT animals implying the possibility of mtDNA mutation recurrence. This work has been funded by the grants from the Burroughs Wellcome Fund, the National Institutes of Health (RO1AG062459 and P51 OD011092), National Research Foundation of Korea (2018R1D1A1B07043216) and Oregon Health & Science University institutional funds. The authors declare no competing interests.

Cytoplasmic inheritance and its effects on development and performance

In contrast to nuclear inheritance, cytoplasmic inheritance in mammals is derived mostly, if not exclusively, from the maternal line. Mitochondria, and their DNA molecules (mtDNA), are the genetic units of this method of inheritance. Mammalian mtDNA codes for 13 enzymes used in the mitochondrial energy-generating pathway, oxidative phosphorylation, 22 tRNAs and two rRNAs. Although all transcripts of mtDNA and their translational products remain in the mitochondria, most proteins used in mitochondria are from nuclear DNA and are imported after synthesis on cytoplasmic ribosomes. Spermatozoa introduce a small number of mitochondria into the cytoplasm of the egg at fertilization, which appear to be digested soon after penetration. Although the paternal contribution of mtDNA to the offspring is not believed to occur in mammals, some interspecific crosses have suggested that it does occur. Experiments with animals derived from reconstituted embryos, using nuclear or cytoplasmic transplantations, suggest that nuclear-mitochondrial interactions are important but not essential in the survival and replication of exogenous mitochondria introduced into the egg. As the levels of heteroplasmy varied in several tissues of animals derived from reconstituted embryos, it is suggested that differential partitioning of mitochondria occurs during embryogenesis. Mitochondrial morphology changes substantially during oogenesis and throughout early cleavage stages. Somatic morphology and normal replication patterns are regained at the blastocyst stage. In pig oocytes and embryos, mitochondria aggregate and are closely associated with endoplasmic reticulum, lipid granules and large vesicles. Although the direct correlation of mitochondrial genes with reproductive traits is still unclear, some human degenerative diseases and performance traits in cattle can be related directly to specific mtDNA polymorphisms. In pigs, reciprocal-cross comparisons have indicated greater offspring parent similarity with dam than sire for lean:fat ratio. A difference was also observed for oxygen consumption and oxidative phosphorylation, but not for anaerobic energy metabolism, in a pig reciprocal-cross experiment. Information on the transmission of mtDNA and its effects on performance will have many implications not only for our understanding of mitochondrial genetics but also for the increased productivity of animals. There are also potential ramifications to the animal cloning industry.

Dynamics of paternal contributions to early embryo development in large animals.

This review focuses on current knowledge of paternal contributions to preimplantation embryonic development with particular emphasis on large animals. Specifically, the included content aims to summarize genomic and epigenomic contributions of paternally expressed genes, their regulation, and chromatin structure that are indispensable for early embryo development. The accumulation of current knowledge will summarize conserved allelic function among species to include functional molecular and genomic studies across large domestic animals in context with reference to founding experimental models.

Sperm competition in golden cuttlefish Sepia esculenta: The impact of mating order and male size

Sperm competition, as the last form of male competition prior to fertilization, is an important component in the reproductive and evolutionary biology of polyandrous species. Golden cuttlefish (Sepia esculenta) is a commercially important annual cephalopod, in which females often mate with multiple males during their mating season and maintain viable sperm in their sperm storage organs for a long time. This multiple-mating system leads to male competition and ultimately sperm competition. To explore the mechanisms of sperm competition in cuttlefish, a laboratory study was conducted using three experimental approaches. First, two similar-sized males were introduced to sequentially mate with a single female (experiment 1), to investigate mating order effects on paternity. Thereafter, a four-male experiment was run (experiment 2), which again tested for mating sequence on paternal contribution. Finally, to test whether male size is a factor affecting paternity, we compared the paternity of two different-sized males that were sequentially mated with a single female (experiment 3). The paternity of the offspring was analyzed using six microsatellite DNA markers on fertilized eggs. The results revealed last-male sperm precedence in all experiments, and no effect of male size was detected. For the two-male experiment, the last male to mate had a progeny share (P2) of 42.86%–61.90%. For the four-male experiment, the last mated male had the highest progeny share (P4) with a proportion of 35.56%–43.75%. Even with differential male sizes in the two-male experiment, the last mated male retained the highest progeny share (P2, 58.33%–91.67%). This study shows a consistent pattern of last-sperm advantage across different experiments and contributes to growing research on cephalopod mating systems. Proper mating schemes should be considered during seedling production for S. esculenta to ascertain efficient resource use and genetic diversity in produced offspring.

Prader-Willi syndrome: reflections on seminal studies and future therapies.

Prader-Willi syndrome (PWS) is caused by the loss of function of the paternally inherited 15q11-q13 locus. This region is governed by genomic imprinting, a phenomenon in which genes are expressed exclusively from one parental allele. The genomic imprinting of the 15q11-q13 locus is established in the germline and is largely controlled by a bipartite imprinting centre. One part, termed the Prader-Willi syndrome imprinting center (PWS-IC), comprises a CpG island that is unmethylated on the paternal allele and methylated on the maternal allele. The second part, termed the Angelman syndrome imprinting centre, is required to silence the PWS_IC in the maternal germline. The loss of the paternal contribution of the imprinted 15q11-q13 locus most frequently occurs owing to a large deletion of the entire imprinted region but can also occur through maternal uniparental disomy or an imprinting defect. While PWS is considered a contiguous gene syndrome based on large-deletion and uniparental disomy patients, the lack of expression of only non-coding RNA transcripts from the SNURF-SNRPN/SNHG14 may be the primary cause of PWS. Patients with small atypical deletions of the paternal SNORD116 cluster alone appear to have most of the PWS related clinical phenotypes. The loss of the maternal contribution of the 15q11-q13 locus causes a separate and distinct condition called Angelman syndrome. Importantly, while much has been learned about the regulation and expression of genes and transcripts deriving from the 15q11-q13 locus, there remains much to be learned about how these genes and transcripts contribute at the molecular level to the clinical traits and developmental aspects of PWS that have been observed.

Evaluation of interactive effects between paternal alcohol consumption and paternal socioeconomic status and environmental exposures on congenital heart defects.

While the maternal risk factors on congenital heart defects (CHDs) have often been assessed, paternal contribution to CHDs, especially the joint effects of paternal risk factors on CHDs remain unknown. This study examined the major impacts of paternal alcohol consumption and its interaction (on multiplicative and additive scales) with paternal socioeconomic status (SES) and environmental exposures on CHDs in China. A population-based case-control study involving 4,726 singleton CHDs cases and 4,726 controls (without any malformation and matched on hospital, gender, and gestational age) was conducted in Guangdong, China, 2004-2014. Information on parental demographics, behavioral patterns, disease/medication, and environmental exposures (3 months before pregnancy) was collected through face-to-face interviews. Conditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) while controlling for all parental factors. Paternal alcohol consumption was associated with an increased OR of CHDs (adjusted OR = 2.87, 95% CI: 2.25-3.65). Additionally, paternal smoking, industry occupation, organic solvent contact, virus infection and antibiotic use, living in rural areas, low household income, and migrant status were significantly associated with CHDs (ORs ranged: 1.42-4.44). Significant additive or multiplicative interactions were observed between paternal alcohol consumption and paternal smoking, industrial occupation, and low income on any CHDs (interaction contrast ratio [ICR] = 4.72, 95% CI: 0.96-8.47] and septal defects (ICRs ranged from 2.04 to 2.79, p < .05). Paternal alcohol consumption and multiple paternal factors were significantly associated with CHDs in China. Paternal smoking and low SES factors modified paternal alcohol consumption-CHDs relationships. Further studies are needed to confirm these findings.

Effects of exposure to a “safe” dose of bisphenol A on male reproductive function and the paternal contribution to the hypothalamic transcriptome profile

The hypothalamus plays key roles in regulating reproduction and energy balance. We determined the transcriptional profile of the hypothalamus (F0 and F1) to identify its relationship with the reproductive function and metabolic phenotypes of rats under BPA exposure and the paternal contribution. Male rats received 35 μg/kg bw/day BPA, 5 μg/kg bw/day ethinylestradiol (EE2) or vehicle via diet. At PND 170, the sexual behaviour of male F0 rats was assessed before mating. The metabolic and reproductive phenotypes and the transcriptional profiles of the testis (F0) and hypothalamus were tested in F0 and F1 rats. Male rats exposed to BPA exhibited no changes in fertility, metabolic phenotypes, sperm quality or serum sex hormones, but none of them ejaculated during the test period. Four differentially expressed genes (DEGs) in the F0 hypothalamus and 51 DEGs in the F0 testis were identified in the BPA group. Paternal BPA and EE2 exposure increased the body weight and food intake of male offspring, and 164 and 3433 DEGs were identified in the hypothalamus of BPA- and EE2-treated rats (F1, male), respectively. The highest-ranked function impaired by BPA and EE2 in the F1 hypothalamus was immune function. BPA and EE2 exposure induced alterations in the metabolic phenotype in F1-generation males, and this effect may be related to the altered inflammatory-related signalling pathways in the hypothalamus but neither treatment affected the F0 hypothalamus. Hundreds of genes in the F1 hypothalamus showed changes upon BPA and EE2 treatment. Concern regarding paternal environmental endocrine disruptor exposure should be raised.

VisEgg: a robust phenotyping tool to assess rainbow trout egg features and viability.

Assessing female fish reproductive success requires a thorough evaluation of egg characteristics, including egg number, size, and variability as well as egg developmental potential through the monitoring of embryo survival after fertilization. While embryonic success relies, at least in part, on paternal contribution, some parameters are strictly related to egg characteristics, one of the main ones being the viability of the egg when released into the water at spawning. It is however not necessarily possible, at least in salmonid fish that lay nontransparent eggs, to separate the different causes of egg/embryo failure. In this context, our aim was (i) to develop a simple and rapid system to capture images of rainbow trout eggs combined with computerized processing of these images to perform a fully automatic individual characterization of egg features including number and size (ii) to estimate unfertilized egg viability through the monitoring of the percentage of eggs that will not survive to water hydration. To evaluate the VisEgg system, unfertilized eggs (approximatively 400 eggs per batch) originating from 105 different females were hydrated in water. After 24 h, a picture of the eggs was obtained using a dedicated shooting system consisting of a light source and a digital single-lens reflex (SLR) camera. An image processing algorithm was developed to allow the automatic detection and separation of the eggs and to perform automatic measurements of egg number and individual egg size. The presence of white egg was used as an indirect measure of egg integrity, the "whitening" being the result of water entry into the egg through the vitelline membrane. These white eggs were therefore considered nonviable, as a result of their lack of physical integrity. Fertilization assays were performed in parallel using a subsample of the same egg batch. Embryonic development was monitored and hatching rate was calculated. A significant correlation between white egg percentage after hydration and hatching rate was observed (Spearman coefficient = -0.557, p < 0.001), in consistency with the fact that nonviable egg will not allow successful embryonic development. In contrast, the percentage of eggs that do not successfully hatch includes egg/embryo failures of different nature including reduced egg viability. Using the VisEgg, we were able to quantify the lack of viability of the eggs separately from the different other events that may occur during fertilization and incubation. the VisEgg is a convenient and reliable tool to obtain individual measures on trout eggs. It can be used to assess not only egg size and egg number but also unfertilized egg viability before fertilization.