Sperm-egg Interaction Research Articles

Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm.

Integrins are transmembrane cell receptors involved in two crucial mechanisms for successful fertilization, namely, mammalian intracellular signaling and cell adhesion. Integrins α6β4, α3β1 and α6β1 are three major laminin receptors expressed on the surface of mammalian cells including gametes, and the presence of individual integrin subunits α3, α6, β1 and β4 has been previously detected in mammalian sperm. However, to date, proof of the existence of individual heterodimer pairs in sperm and their detailed localization is missing. The major conclusion of this study is evidence that the β4 integrin subunit is expressed in mouse sperm and that it pairs with subunit α6; additionally, there is a detailed identification of integrin heterodimer pairs across individual membranes in an intact mouse sperm head. We also demonstrate the existence of β4 integrin mRNAs in round spermatids and spermatogonia by q-RT-PCR, which was further supported by sequencing the PCR products. Using super-resolution microscopy accompanied by colocalization analysis, we located integrin subunits as follows: α6/β4-inner apical acrosomal membrane and equatorial segment; α3, α6/β1, β4-plasma membrane overlaying the apical acrosome; and α3/β1-outer acrosomal membrane. The existence of α6β4, α3β1 and α6β1 heterodimers was further confirmed by proximity ligation assay (PLA). In conclusion, we delivered detailed characterization of α3, α6, β1 and β4 integrin subunits, showing their presence in distinct compartments of the intact mouse sperm head. Moreover, we identified sperm-specific localization for heterodimers α6β4, α3β1 and α6β1, and their membrane compartmentalization and the presented data show a complexity of membranes overlaying specialized microdomain structures in the sperm head. Their different protein compositions of these individual membrane rafts may play a specialized role, based on their involvement in sperm-epithelium and sperm-egg interaction.

Sperm-egg interaction assay for determination of fertile period in ostrich (Struthio camelus)

Sperm-egg interaction assay for determination of fertile period in ostrich (<i>Struthio camelus</i>)

Differential sperm-egg interactions in experimental pairings between two subspecies and their hybrids in a passerine bird.

Speciation research has largely overlooked reproductive barriers acting between copulation and the formation of the zygote (i.e., postmating, prezygotic [PMPZ] barriers), especially in internally fertilizing vertebrates. Nonetheless, it is becoming clear that PMPZ reproductive barriers can play a role in the formation and maintenance of species boundaries. We investigated sperm‐egg interactions in the recently diverged subspecies pairs of the long‐tailed finch, Poephila acuticauda acuticauda and P. a. hecki, to explore potential PMPZ barriers. Specifically, we compared the number of sperm reaching the perivitelline layer (PVL) of the ova, and hence the site of fertilization, in both intra‐ and inter‐subspecies pairings and pairings of F1 hybrid adults with one parental subspecies. Although we found no difference in PVL sperm number among intra‐ and inter‐subspecific pairs, a significantly lower number of sperm reached the site of fertilization in a backcross pairing. As low numbers of PVL sperm appear to be associated with low fertilization success in birds, our findings offer insight into the potential role of postcopulatory processes in limiting gene flow between the subspecies and may help explain the relatively narrow hybrid zone that exists in the wild in this species. Though further work is needed to gain a comprehensive understanding of the morphological, physiological, and molecular mechanisms underlying our results, our study supports the role of PMPZ reproductive barriers in avian speciation, even in recently diverged taxa, that may not yet be fully genetically incompatible.

Identification and characterization of the zebra finch (Taeniopygia guttata) sperm proteome

Spermatozoa exhibit remarkable variability in size, shape, and performance. Our understanding of the molecular basis of this variation, however, is limited, especially in avian taxa. The zebra finch (Taeniopygia guttata) is a model organism in the study of avian sperm biology and sperm competition. Using LC-MS based proteomics, we identify and describe 494 proteins of the zebra finch sperm proteome (ZfSP). Gene ontology and associated bioinformatics analyses revealed a rich repertoire of proteins essential to sperm structure and function, including proteins linked to metabolism and energetics, as well as tubulin binding and microtubule related functions. The ZfSP also contained a number of immunity and defense proteins and proteins linked to sperm motility and sperm-egg interactions. Additionally, while most proteins in the ZfSP appear to be evolutionarily constrained, a small subset of proteins are evolving rapidly. Finally, in a comparison with the sperm proteome of the domestic chicken, we found an enrichment of proteins linked to catalytic activity and cytoskeleton related processes. As the first described passerine sperm proteome, and one of only two characterized avian sperm proteomes, the ZfSP provides a significant step towards a platform for studies of the molecular basis of sperm function and evolution in birds. SignificanceUsing highly purified spermatozoa and LC-MS proteomics, we characterise the sperm proteome of the Zebra finch; the main model species for the avian order Passeriformes, the largest and most diverse of the avian clades. As the first described passerine sperm proteome, and one of only two reported avian sperm proteomes, these results will facilitate studies of sperm biology and mechanisms of fertilisation in passerines, as well as comparative studies of sperm evolution and reproduction across birds and other vertebrates.

RNA-seq coupled to proteomic analysis reveals high sperm proteome variation between two closely related marine mussel species

Speciation mechanisms in marine organisms have attracted great interest because of the apparent lack of substantial barriers to genetic exchange in marine ecosystems. Marine mussels of the Mytilus edulis species complex provide a good model to study mechanisms underlying species formation. They hybridise extensively at many localities and both pre- and postzygotic isolating mechanisms may be operating. Mussels have external fertilisation and sperm cells should show specific adaptations for survival and successful fertilisation. Sperm thus represent key targets in investigations of the molecular mechanisms underlying reproductive isolation. We undertook a deep transcriptome sequencing (RNA-seq) of mature male gonads and a 2DE/MS-based proteome analysis of sperm from Mytilus edulis and M. galloprovincialis raised in a common environment. We provide evidence of extensive expression differences between the two mussel species, and general agreement between the transcriptomic and proteomic results in the direction of expression differences between species. Differential expression is marked for mitochondrial genes and for those involved in spermatogenesis, sperm motility, sperm-egg interactions, the acrosome reaction, sperm capacitation, ATP reserves and ROS production. Proteins and their corresponding genes might thus be good targets in further genomic analysis of reproductive barriers between these closely related species. SignificanceModel systems for the study of fertilization include marine invertebrates with external fertilisation, such as abalones, sea urchins and mussels, because of the ease with which large quantities of gametes released into seawater can be collected after induced spawning. Unlike abalones and sea urchins, hybridisation has been reported between mussels of different Mytilus spp., which thus makes them very appealing for the study of reproductive isolation at both pre- and postzygotic levels. There is a lack of empirical proteomic studies on sperm samples comparing different Mytilus species, which could help to advance this study. A comparative analysis of sperm proteomes across different taxa may provide important insights into the fundamental molecular processes and mechanisms involved in reproductive isolation. It might also contribute to a better understanding of sperm function and of the adaptive evolution of sperm proteins in different taxa. There is now growing evidence from genomics studies that multiple protein complexes and many individual proteins might have important functions in sperm biology and the fertilisation process. From an applied perspective, the identification of sperm-specific proteins could also contribute to the improved understanding of fertility problems and as targets for fertility control.

Gamete-mediated mate choice: towards a more inclusive view of sexual selection.

'Sperm competition'-where ejaculates from two or more males compete for fertilization-and 'cryptic female choice'-where females bias this contest to suit their reproductive interests-are now part of the everyday lexicon of sexual selection. Yet the physiological processes that underlie these post-ejaculatory episodes of sexual selection remain largely enigmatic. In this review, we focus on a range of post-ejaculatory cellular- and molecular-level processes, known to be fundamental for fertilization across most (if not all) sexually reproducing species, and point to their putative role in facilitating sexual selection at the level of the cells and gametes, called 'gamete-mediated mate choice' (GMMC). In this way, we collate accumulated evidence for GMMC across different mating systems, and emphasize the evolutionary significance of such non-random interactions among gametes. Our overall aim in this review is to build a more inclusive view of sexual selection by showing that mate choice often acts in more nuanced ways than has traditionally been assumed. We also aim to bridge the conceptual divide between proximal mechanisms of reproduction, and adaptive explanations for patterns of non-random sperm-egg interactions that are emerging across an increasingly diverse array of taxa.

Beware of memes in the interpretation of your results - lessons from gene-disrupted mice in fertilization research.

For decades, researchers in the fertilization field reported various candidate factors involved in sperm-egg interaction through experiments using enzyme inhibitors and/or antibodies. However, almost all of these factors have been shown to be nonessential by gene disruption experiments. Recently, attention has focused on the low reproducibility of papers in many research fields. In this Review, I retrospectively revisit how fertilization factors were misinterpreted and led to wrong hypotheses in relation to the reportedly low reproducibility of scientific papers.

Evolution of the testis and spermatozoon in mice and rats (Subfamily Murinae) in the absence of sperm competition

AbstractThis study explores the potential effects of interspecific differences in breeding systems on testis organisation and sperm morphology of native murid rodents. It poses the question – what are the effects of depressed levels of intermale sperm competition, as indicated by small relative testes mass, on the morphology of testes and spermatozoa in murine rodents? Species from three separate murine tribes, those of the Hydromyini, Rattini and Arvicanthini, were investigated with low relative testes mass being used as a proxy for low, or non‐existent, levels of intermale sperm competition. The findings show that in only one of the tribes with low relative testes mass, that of the Hydromyini, was there a reduced testicular area occupied by seminiferous tubules, but in two of the tribes significantly smaller seminiferous tubule diameters were present. In all three tribes, most species with low relative testes mass had a highly derived sperm morphology that, unlike in species with large relative testes mass, had spermatozoa where the head lacked an apical hook, was highly variable in form, often had a very large acrosome overlying the apical region of the nucleus which suggests divergent processes of sperm–egg interaction at the time of fertilisation, and a significantly shorter tail. It is therefore hypothesised that, unlike in species with a large relative testes mass where high levels of intermale sperm competition maintain a streamlined sperm head with an apical hook to aid in zona penetration, in most species of murine rodents with low relative testes mass there is greater reliance on enzymatic digestion of the extracellular matrix around the egg, and especially that of the zona pellucida, to facilitate sperm penetration at the time of fertilisation.

Assessment of azadirachtin-A, a neem tetranortritarpinoid, on rat spermatozoa during in vitro capacitation.

The exploration of the biological assessment of technical azadirachtin, a tetranortritarpinoid from the neem seed kernel, was reviewed. The present study was, therefore, designed to evaluate the dose-dependent in vitro effects of azadirachtin-A, particularly on the functional studies and determination of molecular events, which are critical in the process of sperm capacitation. To assess the effects of the azadirachtin-A on the functional studies, sperm capacitation, the total sperm adenosine triphosphate levels, acrosome reaction (AR), the sperm-egg interaction and the determination of molecular events like cyclic adenosine-3',5'-monophosphate and calcium levels, the appropriate volumes of the sperm suspension were added to the medium to a final concentration of 1×106 sperm/mL and incubated in a humidified atmosphere of 5% CO2 in air at 37°C. The increasing quantities 0.5-2.0 mM/mL and the equivalent volumes of 50% dimethyl sulfoxide were added to the control dishes prior to the addition of spermatozoa and then observed at various time-points for motility and other analyses. Results revealed the dose- and time-dependent decrease in the functional consequence of capacitation, i.e. the percentage of motile spermatozoa, motility score and sperm motility index, levels of molecular events in spermatozoa, followed by declined spontaneous AR leading to lesser binding of the cauda epididymal sperm to the Zona pellucida. The findings confirm the inhibition of rat sperm motility by blocking some biochemical pathways like energy utilization. They also demonstrate that sperm capacitation is associated with the decrease in AR and that the levels of molecular events in spermatozoa can guide us towards the development of a new male contraceptive constituent.

Sperm morphology and the evolution of intracellular sperm-egg interactions.

Sperm morphology is incredibly diverse, even among closely related species, yet the coevolution between males and females of fertilization recognition systems is necessary for successful karyogamy (male and female pronuclear fusion). In most species, the entire sperm enters the egg during fertilization so sperm morphological diversity may impact the intracellular sperm–egg interactions necessary for karyogamy. We quantified morphological variation of sperm inside eggs prior to and following karyogamy in several species of Drosophila to understand whether evolution of sperm morphology could influence intracellular sperm–egg interactions (ISEIs). We measured seven parameters that describe ISEIs among species to determine whether these parameters varied both within a species across development and across species at the same developmental stage. We used heterospecific crosses to test the relative role of male origin, female origin, and interaction between the male and female in determining ISEIs. We found that sperm shape changed within a species as development proceeded and, at particular development stages, species varied in some ISEIs. Parental origin had an effect on some ISEIs, with a general trend for a stronger female effect. Overall, our findings identify conserved and variable ISEIs among species and demonstrate the potential to contribute understanding to gamete evolution and development.

A New Model for Understanding the Egg Cell Surface at Fertilization

The cell signaling pathway initiating fertilization has not been fully elucidated in any species. Several downstream components in the pathway leading to an internal Ca2+ release have been found; however, not much is known regarding the participation of proteins on the surface of the sperm and egg. The starfish Patiria miniata is an excellent model organism to study this process because they produce a significant quantity of large synchronized oocytes that are nearly transparent and rigid enough to hold their spherical shape despite the removal of extracellular layers. Recently, lipid rafts have been found on the surface of a variety of eggs, but their role at fertilization is unknown. Lipid rafts are small dynamic regions enriched with cholesterol and sphingolipids, which hold clusters of membrane bound proteins. Lipid rafts are found in a variety of cells and they are critical in certain signaling processes, such as the activation of T cells. This study develops the starfish egg as a model system for the study of lipid rafts and for the discovery of cell surface proteins necessary for fertilization. To visualize egg surface proteins in living cells, oocytes were labeled with a membrane impermeant form of biotin and imaged during fertilization by laser scanning confocal microscopy. Evidence of an increase in protein at the fertilization cone, which marks the site of sperm‐egg fusion, was seen. To further understand how these surface proteins were behaving at fertilization, samples prepared from the biotinylated eggs were subjected to gel electrophoresis followed by affinity interaction with streptavidin‐HRP to visualize the labeled proteins. This showed the presence of multiple egg surface proteins, ranging in size from 75 to 250 kDa in great quantity, as well as a few proteins ranging from 30 to 75kDa. Interestingly, biotinylated proteins were also recovered from the surrounding seawater following fertilization, suggesting the action of a protease that cleaves egg cell surface proteins. Experiments with protease inhibitors suggest that this protease activity is necessary for fertilization, leading to the idea that the protease may be involved in sperm‐egg fusion and/or signaling. To examine the idea that signaling may be initiated at the moment of sperm‐egg interaction, and to determine if lipid rafts could be involved, a detergent‐based lipid raft isolation assay was developed for the starfish egg. The eggs are separated into cytosolic, soluble membrane, and insoluble membrane (lipid rafts) fractions. The fractions were subjected to western blotting for tyrosine‐phosphorylated proteins, which were found almost exclusively in the insoluble fraction of the unfertilized egg. Therefore, the lipid rafts contain as of yet unidentified signaling molecules that could play a role in the initiation of fertilization. Understanding this mechanism at the molecular level in starfish can to eventually lead to understanding infertility issues as well as offer new possible non‐hormonal prophylactics in humans.Support or Funding InformationThis research was funded by the NSF and Florida Institute of TechnologyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Sperm-egg interaction and fertilization: past, present, and future.

Fifty years have passed since the findings of capacitation and acrosome reaction. These discoveries and the extensive effort of researchers led to the success of in vitro fertilization, which has become a top choice for patients at infertility clinics today. The effort to understand the mechanism of fertilization is ongoing, but the small number of eggs and similarly small quantity of spermatozoa continue to hinder biochemical experiments. The emergence of transgenic animals and gene disruption techniques has had a significant effect on fertilization research. Factors considered important in the early years were shown not to be essential and were replaced by newly found proteins. However, there is much about sperm-egg interaction which remains to be learned before we can outline the mechanism of fertilization. In fact, our understanding of sperm-egg interaction is entering a new stage. Progress in transgenic spermatozoa helped us to observe the behavior of spermatozoa in vivo and/or at the moment of sperm-egg fusion. These advancements are discussed together with the paradigm-shifting research in related fields to help us picture the direction which fertilization research may take in the future.

Epididymal cysteine-rich secretory proteins are required for epididymal sperm maturation and optimal sperm function.

What is the role of epididymal cysteine-rich secretory proteins (CRISPs) in male fertility? While epididymal CRISPs are not absolutely required for male fertility, they are required for optimal sperm function. CRISPs are members of the CRISP, Antigen 5 and Pathogenesis related protein 1 (CAP) superfamily and are characterized by the presence of an N-terminal CAP domain and a C-terminal CRISP domain. CRISPs are highly enriched in the male reproductive tract of mammals, including in the epididymis. Within humans there is one epididymal CRISP, CRISP1, whereas in mice there are two, CRISP1 and CRISP4. In order to define the role of CRISPs within the epididymis, Crisp1 and Crisp4 knockout mouse lines were produced then interbred to produce Crisp1 and 4 double knockout (DKO) mice, wherein the expression of all epididymal CRISPs was ablated. Individual and DKO models were then assessed, relative to their own strain-specific wild type littermates for fertility, and sperm output and functional competence at young (10-12 weeks of age) and older ages (22-24 weeks). Crisp1 and 4 DKO and control mice were also compared for their ability to bind to the zona pellucida and achieve fertilization. Knockout mouse production was achieved using modified embryonic stem cells and standard methods. The knockout of individual genes was confirmed at a mRNA (quantitative PCR) and protein (immunochemistry) level. Fertility was assessed using breeding experiments and a histological assessment of testes and epididymal tissue. Sperm functional competence was assessed using a computer assisted sperm analyser, induction of the acrosome reaction using progesterone followed by staining for acrosome contents, using immunochemical and western blotting to assess the ability of sperm to manifest tyrosine phosphorylation under capacitating conditions and using sperm-zona pellucida binding assays and IVF methods. A minimum of three biological replicates were used per assay and per genotype. While epididymal CRISPs are not absolutely required for male fertility, their production results in enhanced sperm function and, depending on context, CRISP1 and CRISP4 act redundantly or autonomously. Specifically, CRISP1 is the most important CRISP in the establishment of normally motile sperm, whereas CRISP4 acts to enhance capacitation-associated tyrosine phosphorylation, and CRISP1 and CRISP4 act together to establish normal acrosome function. Both are required to achieve optimal sperm-egg interaction. The presence of immune infiltrates into the epididymis of older, but not younger, DKO animals also suggests epididymal CRISPs function to produce an immune privileged environment for maturing sperm within the epididymis. Caution should be displayed in the translation of mouse-derived data into the human wherein the histology of the epididymis is someone what different. The mice used in the study were housed in a specific pathogen-free environment and were thus not exposed to the full range of environmental challenges experienced by wild mice or humans. As such, the role of CRISPs in the maintenance of an immune privileged environment, for example, may be understated. The combined deletion of Crisp1 and Crisp4 in mice is equivalent to the removal of all CRISP expression in humans. As such, these data suggest that mammalian CRISPs, including that in humans, function to enhance sperm function and thus male fertility. These data also suggest that in the presence of an environmental challenge, CRISPs help to maintain an immune privileged environment and thus, protect against immune-mediated male infertility. Not applicable. This study was funded by the National Health and Medical Research Council, the Victorian Cancer Agency and a scholarship from the Chinese Scholarship Council. The authors have no conflicts of interest to declare.

Gynura procumbens Improved Fertility of Diabetic Rats: Preliminary Study of Sperm Proteomic.

Gynura procumbens (GP) is a medicinal herb that has long been known as anti-inflammatory and antihyperglycaemic. Recently, this herbal extract has been associated with a profertility effect, suggesting its applicability in treating both diabetes and male infertility. In this study, the effects of GP aqueous extract (GPAE) on diabetic rats were investigated through evaluating testes histology and androgen hormone levels as well as the implantation sites of female rats on copulation with the treated male rats. Three dosages of GPAE were used (150, 300, and 450 mg/kg), and there were three control groups [normal, diabetic, and metformin-treated diabetic]. Testes histology, androgen hormone levels, and number of implantation sites of the GPAE-treated groups matched those of the normal group in contrast to the diabetic and metformin-treated diabetic controls. Sperm proteomics analysis identified 666 proteins, but only 88 were consistently found in all the control and 450-mg/kg GPAE-treated groups. Four proteins, including cysteine-rich secretory protein 1, carboxylesterase 5A, zona pellucida binding protein, and phosphatidylethanolamine-binding protein 1, were significantly upregulated with GPAE treatment compared with the diabetic control, matching the protein levels of the normal group. These proteins were mainly involved in sperm maturation, sperm capacitation, and sperm-egg interaction, suggesting that GP treatment was able to restore the fertility of male diabetic rats at molecular protein level. In conclusion, GP treatment effectively treats infertility of male diabetic rats, possibly through the upregulation of proteins related to sperm maturation and sperm-egg interaction.

De novo male gonad transcriptome draft for the marine mussel Perumytilus purpuratus with a focus on its reproductive-related proteins.

Perumytilus purpuratus is a marine mussel considered a bioengineer species with a broad distribution in the Pacific and Atlantic coast of South America. Studies have shown two geographically and genetically differentiated subpopulations at molecular level and in sperm morphological traits. To open avenues for molecular research on P. purpuratus, a global de novo transcriptome from gonadal tissue of mature males was sequenced using the Illumina platform. From a total of 126.38 million reads, 37,765 transcripts were successfully annotated. BUSCO analysis determined a level of 89% completeness for the assembled transcriptome. The functional gene ontology (GO) annotation indicated that, in terms of abundance, the transcripts related with molecular function were the most represented, followed by those related with biological process and cellular components. Additionally, a subset of GO annotations generated using the “sperm” term resulted in a total of 1,294 sequences where the biological process category was the more represented, with transcripts strongly associated to sperm-processes required for fertilization, and with processes where the sperm-egg interaction could be implicated.Our work will contribute to the evolutionary understanding of the molecular mechanisms related to tissue-specific functions. This work reports the first male gonad transcriptome for the mussel P. purpuratus, generating a useful transcriptomic resource for this species and other closely related mytilids.

Factors controlling sperm migration through the oviduct revealed by gene-modified mouse models.

Mammalian fertilization is comprised of many steps including sperm survival in the uterus, sperm migration in the female reproductive tract, physiological and morphological changes to the spermatozoa, and sperm-egg interaction in the oviduct. In vitro studies have revealed essential factors for these fertilization steps for over half a century. However, the molecular mechanism of fertilization has recently been revised by the emergence of genetically modified animals. Here, we focus on essential factors for sperm fertilizing ability and describe recent advances in our knowledge of the mechanisms of mammalian fertilization, especially of sperm migration from the uterus into the oviduct.

Male-female relatedness at specific SNP-linkage groups influences cryptic female choice in Chinook salmon (Oncorhynchus tshawytscha).

In a range of taxa, the relatedness between mates influences both pre- and post-mating processes of sexual selection. However, relatively little is known about the genetic loci facilitating such a bias, with the exception of the major histocompatibility complex. Here, we performed tightly controlled replicated in vitro fertilization trials to explore the impact of relatedness on two possible mechanisms of cryptic female choice (CFC) in Chinook salmon (Oncorhynchus tshawytscha). We tested (i) whether relatedness of mates, assessed using 682 single nucleotide polymorphisms (SNPs) on 29 SNP-linkage groups (LGs), biases a male's sperm velocity in ovarian fluid (a parameter previously shown to predict male fertilization success), and (ii) whether relatedness of mates governs fertilization success via other mechanisms, probably via sperm-egg interactions. We found that relatedness on three LGs explained the variation in sperm velocity, and relatedness on two LGs explained fertilization success, which might indicate the presence of genes important in sperm-ovarian fluid and sperm-egg interactions in these genomic regions. Mapping of the SNPs on these LGs to the rainbow trout genome revealed two genes that affect fertility in humans and represent candidate genes for further studies. Our results thereby provide a novel contribution to the understanding of the mechanism of CFC.

In vitro evidence for the participation of Drosophila melanogaster sperm β‐N‐acetylglucosaminidases in the interactions with glycans carrying terminal N‐acetylglucosamine residues on the egg's envelopes

Fertilization is a complex and multiphasic process, consisting of several steps, where egg-coating envelope's glycoproteins and sperm surface receptors play a critical role. Sperm-associated β-N-acetylglucosaminidases, also known as hexosaminidases, have been identified in a variety of organisms. Previously, two isoforms of hexosaminidases, named here DmHEXA and DmHEXB, were found as intrinsic proteins in the sperm plasma membrane of Drosophila melanogaster. In the present work, we carried out different approaches using solid-phase assays in order to analyze the oligosaccharide recognition ability of D. melanogaster sperm hexosaminidases to interact with well-defined carbohydrate chains that might functionally mimic egg glycoconjugates. Our results showed that Drosophila hexosaminidases prefer glycans carrying terminal β-N-acetylglucosamine, but not core β-N-acetylglucosamine residues. The capacity of sperm β-N-acetylhexosaminidases to bind micropylar chorion and vitelline envelope was examined in vitro assays. Binding was completely blocked when β-N-acetylhexosaminidases were preincubated with the glycoproteins ovalbumin and transferrin, and the monosaccharide β-N-acetylglucosamine. Overall, these data support the hypothesis of the potential role of these glycosidases in sperm-egg interactions in Drosophila.

Deficiency in Sperm-Egg Protein Interaction as a Major Cause of Fertilization Failure.

Complete elucidation of fertilization process at molecular level is one of the unresolved challenges in sexual reproduction studies, and understanding the molecular mechanism is crucial in overcoming difficulties in infertility and unsuccessful in vitro fertilization. Sperm-oocyte interaction is one of the most remarkable events in fertilization process, and deficiency in protein-protein interactions which mediate this interaction is a major cause of unexplained infertility. Due to detection of how the various defects of sperm-oocyte interaction can affect fertilization failure, different experimental methods have been applied. This review summarizes the current understanding of sperm-egg interaction mechanism during fertilization and also accumulates the different types of sperm-egg interaction abnormalities and their association with infertility. Several detection approaches regarding sperm-egg protein interactions and the associated defects are reviewed in this paper.

Sensitivity of sea urchin fertilization to pH varies across a natural pH mosaic.

In the coastal ocean, temporal fluctuations in pH vary dramatically across biogeographic ranges. How such spatial differences in pH variability regimes might shape ocean acidification resistance in marine species remains unknown. We assessed the pH sensitivity of the sea urchin Strongylocentrotus purpuratus in the context of ocean pH variability. Using unique male–female pairs, originating from three sites with similar mean pH but different variability and frequency of low pH (pHT ≤ 7.8) exposures, fertilization was tested across a range of pH (pHT 7.61–8.03) and sperm concentrations. High fertilization success was maintained at low pH via a slight right shift in the fertilization function across sperm concentration. This pH effect differed by site. Urchins from the site with the narrowest pH variability regime exhibited the greatest pH sensitivity. At this site, mechanistic fertilization dynamics models support a decrease in sperm–egg interaction rate with decreasing pH. The site differences in pH sensitivity build upon recent evidence of local pH adaptation in S. purpuratus and highlight the need to incorporate environmental variability in the study of global change biology.