Reproductive Fluids Research Articles

Update on Trichomoniasis

In recent years diagnostic surveillance for bovine trichomoniasis has increased since most states require testing prior to interstate movement of non-virgin bulls. However, bovine trichomoniasis continues to be an economically significant disease, particularly in beef cattle, in much of the United States. The disease produces no clinical signs in bulls but can have devastating reproductive consequences on females resulting in reduced calf crops and extended calving seasons. The organism is efficiently transmitted via the venereal route. Although prevalence may be highest in older bulls it has been documented to occur in non-virgin bulls of any age. Bulls are believed to be persistently infected but females develop short lived immunity and naturally clear the infection within 3-22 months. Diagnosis is made by culture in selective media and/or by PCR identification of the organism in smegma samples collected by preputial scraping. Samples of reproductive or fetal fluids obtained from females can also be tested. Readers should consult their state veterinary diagnostic labs for instructions on sample preparation and shipment.
 Control of trichomoniasis is primarily through testing and culling of positive bulls. There are no approved, effective treatments available. However, vaccination of females will, in most cases, improve reproductive indices in infected herds.

Sperm Proteome after Interaction with Reproductive Fluids in Porcine: From the Ejaculation to the Fertilization Site.

Ejaculated sperm are exposed to different environments before encountering the oocyte. However, how the sperm proteome changes during this transit remains unsolved. This study aimed to identify proteomic changes in boar sperm after incubation with male (seminal plasma, SP) and/or female (uterine fluid, UF; and oviductal fluid, OF) reproductive fluids. The following experimental groups were analyzed: (1) SP: sperm + 20% SP; (2) UF: sperm + 20% UF; (3) OF: sperm + 20% OF; (4) SP + UF: sperm + 20% SP + 20% UF; and (5) SP+OF: sperm + 20% SP + 20% OF. The proteome analysis, performed by HPLC-MS/MS, allowed the identification of 265 proteins. A total of 69 proteins were detected in the UF, SP, and SP + UF groups, and 102 proteins in the OF, SP, and SP + OF groups. Our results showed a higher number of proteins when sperm were incubated with only one fluid than when they were co-incubated with two fluids. Additionally, the number of sperm-interacting proteins from the UF group was lower than the OF group. In conclusion, the interaction of sperm with reproductive fluids alters its proteome. The description of sperm-interacting proteins in porcine species after co-incubation with male and/or female reproductive fluids may be useful to understand sperm transport, selection, capacitation, or fertilization phenomena.

Reproductive Outcomes and Endocrine Profile in Artificially Inseminated versus Embryo Transferred Cows.

The increasing use of in vitro embryo production (IVP) followed by embryo transfer (ET), alongside with cryopreservation of embryos, has risen concerns regarding the possible altered pregnancy rates, calving or even neonatal mortality. One of the hypotheses for these alterations is the current culture conditions of the IVP. In an attempt to better mimic the physiological milieu, embryos were produced with female reproductive fluids (RF) as supplements to culture medium, and another group of embryos were supplemented with bovine serum albumin (BSA) as in vitro control. Embryos were cryopreserved and transferred while, in parallel, an in vivo control (artificial insemination, AI) with the same bull used for IVP was included. An overview on pregnancy rates, recipients' hormonal levels, parturition, and resulting calves were recorded. Results show much similarity between groups in terms of pregnancy rates, gestation length and calves' weight. Nonetheless, several differences on hormonal levels were noted between recipients carrying AI embryos especially when compared to BSA. Some calving issues and neonatal mortality were observed in both IVP groups. In conclusion, most of the parameters studied were similar between both types of IVP derived embryos and the in vivo-derived embryos, suggesting that the IVP technology used was efficient enough for the safe production of calves.

Determination of urinary carnitine levels as a potential indicator of uterine fibroids caused by nonylphenol exposure

Our previous studies have shown that uterine fibroids are associated with nonylphenol (NP) exposure, and the changes of carnitines in critical reproductive tissues and body fluids could be used to indicate the female reproductive toxicity caused by NP exposure. In this work, on the basis of further clarifying the correlation between NP exposure level and uterine fibroids, the possibility of the urinary carnitine levels as a potential indicator of uterine fibroids caused by NP exposure was discussed. The urine samples were collected from 84 female volunteers: the control group of 34 healthy women without gynecological disease and 50 uterine fibroids patients, respectively. Methods were respectively established for the determination of NP and eight carnitines in human urine samples by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). The results showed that the NP level of uterine fibroids group was significantly higher than that of control group (P = 0.002), indicating that NP exposure was an important environmental factor in the occurrence of uterine fibroids. It was further found that in urine samples of the uterine fibroids group, the levels of L-Carnitine (C0), L-Acetyl-carnitine (C2), L-Octanoyl-carnitine (C8), Tetradecanoyl-carnitine (C14), Oleoyl-carnitine (C18:1) and Linoleoyl-carnitine (C18:2) had obviously increased compared with those in the control group (P < 0.001; < 0.001; < 0.001; = 0.003; < 0.001; = 0.010). The concentrations of L-Hexanoyl-carnitine (C6) and L-Palmitoyl-carnitine (C16) in the uterine fibroids group were also higher than those in the control group, although the difference was not statistically significant (P > 0.05). The results suggested that the changes in urinary carnitine levels might be a potential indicator to help to warn of the risk of uterine fibroids caused by NP exposure at the early stage.

Microbiota and Malodor-Etiology and Management.

Accumulating evidence indicates that microbiota plays a critical role in physiological processes in humans. However, it might also contribute to body malodor by producing numerous odorous molecules such as ammonia, volatile sulfur compounds or trimethylamine. Although malodor is commonly overlooked by physicians, it constitutes a major problem for many otherwise healthy people. Thus, this review aims to investigate most common causes of malodor and describe potential therapeutic options. We searched PUBMED and Google Scholar databases to identify the clinical and pre-clinical studies on bad body smell, malodor, halitosis and microbiota. Unpleasant smell might originate from the mouth, skin, urine or reproductive fluids and is usually caused by odorants that are produced by resident bacterial flora. The accumulation of odorous compounds might result from diet, specific composition of microbiota, as well as compromised function of the liver, intestines and kidneys. Evidence-based guidelines for management of body malodor are lacking and no universal treatment exists. However, the alleviation of the symptoms may be achieved by controlling the diet and physical elimination of bacteria and/or accumulated odorants.

The dog as a sentinel species for environmental effects on human fertility.

Despite the vast body of evidence that environmental toxicants adversely affect reproductive development and function across species, demonstrating true cause and effect in the human remains a challenge. Human meta-analytical data, showing a temporal decline in male sperm quality, are paralleled by a single laboratory study showing a similar 26-year decline in the dog, which shares the same environment. These data are indicative of a common cause. Environmental chemicals (ECs) detected in reproductive tissues and fluids induce similar, short term, adverse effects on human and dog sperm. Both pre- and post-natal stages of early life development are sensitive to chemical exposures and such changes could potentially cause long term effects in the adult. The environmental 'pollutome' (mixtures of ECs) is determined by industrialisation, atmospheric deposition and bioaccumulation and characterises real-life exposure. In Arctic ecosystems, dietary and non-dietary chemical contaminants are detectable in biological tissues and linked with adverse health effects in both dogs and their handlers. In the female, such exposure could contribute to disorders such as ovarian insufficiency, dysregulated follicle development, ovarian cancer, and polycystic ovarian syndrome. In the dog, ovarian chemical concentrations are greater in the testis. In addition, preliminary studies indicate that dietary exposures may influence the sex ratio in the offspring in favour of females. Within this article, we review current knowledge on chemical effects on human reproduction and suggest that the dog, as a sentinel species for such effects, is an essential tool for addressing critical data gaps in this field.

Partitioning of fatty acids into tissues and fluids from reproductive organs of ewes as affected by dietary phenolic extracts

Evaluation of dietary interventions with regard to fertility problems often observed in ruminant livestock is of global interest. Though the effects of polyphenol supplementation in ruminants on digestion and food quality are well described, the impact on reproductive tissues and fluids remains scarcely investigated. These compounds protect dietary unsaturated fatty acids (FA) from oxidation and biohydrogenation and thus saturation. In addition, modification of the expression of genes associated with FA metabolism may occur. Therefore, we characterized for the first time the FA profiles of reproductive tissues and fluids and investigated their potential modification by dietary polyphenols in 22 cyclic ewes. The animals were randomly divided into four groups and fed a basal diet of meadow hay and one of four concentrate types either non-supplemented (control) or supplemented with grape seed extract, Acacia mearnsii bark extract (13 g/kg dry matter (DM) each) or a combination of both (26 g/kg DM). After 10 weeks of feeding, the animals were slaughtered. Samples of reproductive (oviduct, uterus) and metabolically differently active tissues (liver, muscle, adipose) as well as of plasma and fluids from oviduct and uterus were analysed for their FA composition. In addition, the expression of lipid metabolic and antioxidant genes was analysed in all tissues except the adipose tissue. Fatty acid profiles in tissues and fluids as well as gene expression in tissues significantly differed between the different fluids and tissues. In contrast, only a few diet and matrix (fluid or tissue) × diet interactions were observed. Still, the FA profile of the uterus was the only one not at all affected by the diet. The mRNA expression was not affected by the diet for most of the genes investigated, which might in part be explained by the similar plasma polyphenol concentrations found at slaughter. Overall, our findings contribute to an improved understanding of the characteristic FA composition of reproductive tissues and fluids in sheep. In addition, the effect of polyphenols on different tissues, fluids and tissue gene expression has been confirmed as described in other animal species.

Thiol based mechanism internalises interacting partners to outer dense fibers in sperm

The sperm tail outer dense fibres (ODFs) contribute passive structural role in sperm motility. The level of disulphide cross-linking of ODFs and their structural thickness determines flagellar bending curvature and motility. During epididymal maturation, proteins are internalized to modify ODF disulphide cross-linking and enable motility. Sperm thiol status is further altered during capacitation in female tract. This suggests that components in female reproductive tract acting on thiol/disulphides could be capable of modulating the tail stiffness to facilitate modulation of the sperm tail rigidity and waveform en route to fertilization. Understanding the biochemical properties and client proteins of ODFs in reproductive tract fluids will help bridge this gap. Using recombinant ODF2 (aka Testis Specific Antigen of 70 kDa) as bait, we identified client proteins in male and female reproductive fluids. A thiol-based interaction and internalization indicates sperm can harness reproductive tract fluids for proteins that interact with ODFs and likely modulate the tail stiffness en route to fertilization.

Periovulatory oviductal fluid decreases sperm protein kinase A activity, tyrosine phosphorylation, and in vitro fertilization in pig.

Molecules from the female reproductive tract modulate capacitation and function of sperm cells in vivo. These molecules vary in a quantitative and qualitative manner throughout the estrous cycle. This work evaluates the effect of using various female reproductive fluids on capacitation and fertilization of pig spermatozoa in vitro. The effects of culturing spermatozoa in different fluids on the levels of sperm protein kinase A (pPKA), tyrosine phosphorylation, acrosome reaction, and in vitro fertilization (IVF) were evaluated. The fluids tested were as follows: oviductal fluid (OF) from five phases of the estrous cycle, namely early and late follicular (OF-EF, OF-LF), early and late luteal (OF-EL, OF-LL) and periovulatory (pOF), follicular fluid from medium-sized follicles, and secretions of cumulus-oocyte complexes (conditioned medium). The pPKAs and tyrosine phosphorylation were decreased by OF-EF, OF-LF, OF-EL, and pOF but not by follicular fluid and conditioned medium. OF-EF, OF-LF, and pOF also decreased the sperm acrosome reaction. Moreover, the effect of pOF on pPKAs and tyrosine phosphorylation was reversible. In in vitro fertilization, OF-EF, OF-LF, OF-EL, and pOF reduced the percentage of penetrated oocytes, the mean number of spermatozoa per penetrated oocyte, and increased monospermy. OF from follicular, early luteal, and periovulatory phases of the estrous cycle modulates the sperm protein phosphorylation as well as the acrosome reaction involved in capacitation and increases monospermic fertilization in in vitro fertilization. Our findings suggest that fluids from the female reproductive tract could be used as additives in porcine IVF systems to modulate sperm-oocyte interaction.

Going beyond conventional parameters to unveil sperm quality in fish: the use of fibre optic technology to assess mitochondrial respiratory performance.

ABSTRACTSperm fertilisation success depends on both intrinsic quality and the interactions with the surrounding reproductive fluids. In several fish species, these interactions have a variable effect on sperm performance. Although specific responses to reproductive fluids may depend on intrinsic differences in sperm quality, variations in the traditionally recorded sperm functional traits do not fully account for the observed patterns. New methods to enhance the evaluation of sperm quality may prove to be valuable at both applied and theoretical levels, by improving the breeding protocol of reared species and the understanding of mating success in sperm competition contexts. Here we develop a fibre optic-based technique, also adequate for small ejaculate samples, to test the role of mitochondrial respiratory efficiency in deciphering sperm performance variability. We purposely used as model the grass goby, Zosterisessor ophiocephalus, a fish with guard-sneaker mating tactics where the sperm in each male tactic have similar intrinsic qualities (velocity, viability, ATP content), but sneakers' sperm exploit territorial males' seminal fuid, overall displaying better fertilization ability. We found that sperm differed in their mitochondrial respiratory efficiency, which was higher in sneakers’ sperm compared to territorial ones. This result draws the attention to an indicator of sperm quality that might be helpful in disentangling the mechanisms driving sperm-reproductive fluid interactions.

Alpine and lowland grazing differentially alter the reproductive tract redox milieu and amino acid composition in cattle

An alpine environment is unique due to pasture biodiversity, with an abundant content of natural antioxidant polyphenols. The present study investigated the effects of lowland and alpine grazing on the oviduct and uterine tissue redox status and amino acid concentrations in plasma and reproductive fluids. In the first experiment, heifers grazed on lowland (H-LOW: n = 13) and on alpine (H-ALP: n = 15) pastures. In the second experiment, heifers grazed on the same lowland (HS-LOW: n = 6) and on a different alpine (HS-ALP: n = 6) pasture. The abundance of mRNA transcripts for antioxidant enzymes in the oviduct (glutathione S-transferase alpha 2, glutathione synthetase (GSS)) and the endometrium (catalase, glutathione-disulfide reductase, GSS) was less (P < 0.05), and for glutathione peroxidase 4 in the endometrium greater (P = 0.006) in the H-LOW than in the H-ALP group. The abundance of mRNA transcript for catalase was less in the endometrium in the H-LOW than in the H-ALP (P = 0.001) group. Catalase and NAD(P)H quinone dehydrogenase 1 concentrations in the oviduct were greater in the HS-LOW than in the HS-ALP group (P < 0.05). Of 32 amino acids analysed, there were differences in concentrations in the H-LOW and H-ALP group of 13, seven and 15 in plasma, oviduct and uterine fluids, respectively (P < 0.05). Comparing the HS-LOW to the HS-ALP groups, there were 13, one and three amino acids in the plasma, oviduct and uterine fluids, respectively, that were differentially abundant (P < 0.05). The grazing systems had some effect on the redox status and amino acid patterns in reproductive tissues.

106 Comparative growth rates and haematological parameters from calves born by transfer of vitrified invitro-produced embryos and stepbrother calves born by AI

Assisted reproductive technologies (ART) are being extensively used to produce cattle offspring. However, as shown by Siqueira et al. (2017 J. Dairy Sci. 100, 5899-5908; https://doi.org/10.3168/jds.2016-12539), phenotypical and performance differences between cows derived from distinct ART can be found at different stages of development. Thus, in an attempt to mimic the natural environment, reproductive fluids (oviductal (SOF) and uterine fluids) were added as supplementation to embryo culture media. Our hypothesis was that this improved culture media would produce calves more similar to the ones produced by AI. Invitro-produced (IVP) beef embryos were produced using SOF media supplemented with reproductive fluids (RF) or standard protocol (BSA), vitrified and later warmed and transferred to synchronized dairy recipients. Simultaneously, other dairy recipients were inseminated (AI) with the same bull used to produce IVP embryos. A total of 19 calves are included in this study (RF n=5, BSA n=7, AI n=7). Calves that did not reach 45 days of life were excluded from these data. All animals received the same feeding and housing conditions. Calves were examined at Days 0, 3, 7, 15, 30, and 45 of life. Each examination included weight, height at withers, thorax circumference, heart and respiratory rates, body temperature, and a blood sample from the jugular vein to perform a general haematological analysis (Siemens ADVIA 120) and glucose levels. A non-parametric test (Mann-Whitney U) was used to compare paired samples, with significance assumed when P&amp;lt;0.05. Since day is a factor that influences growth, it was assumed as a fixed factor, and data were analysed per day. In terms of growth development, AI calves were significantly taller than BSA calves in all days, and in general taller than RF calves, with the exception of Days 3 and 7. Thorax circumference was significantly smaller for BSA versus AI calves only on Days 15 and 45. Respiratory rate was higher for RF calves at birth and for BSA calves at Day 3 when both were compared with AI calves, but we found no difference between them. Heart rate was higher for RF calves on Day 7 compared with BSA and AI, and higher again on Day 15 compared with AI. Regarding haematological parameters, significant differences were found on Day 0, with platelet counts being lower for BSA calves. On Day 7, mean corpuscular volume from AI calves was lower than either BSA or RF calves, and on Day 15, eosinophils were lower for RF calves compared with AI. At Day 30, white blood cells and lymphocyte concentration were lower for BSA than for AI calves. Glucose levels were higher for RF calves than for AI calves on Day 45. Overall, all haematology and clinical values seem to match the values of healthy calves (Brun-Hansen et al. 2006 Vet. Clin. Pathol. 35, 182-187; https://doi.org/10.1111/j.1939-165X.2006.tb00111.x), and the differences found were not clinically relevant. In conclusion, at the moment and from the analysed criteria of development during the first 45 days of life, there seems to exist no difference between calves born by IVP with RF as supplement to culture media and their invitro or invivo controls.

Relationship between concentrations of macro and trace elements in serum and follicular, oviductal, and uterine fluids of the dromedary camel (Camelus dromedarius).

This study aimed at investigating the relationship between concentrations of macro and trace elements in blood serum, and fluids from small and large follicles (SFF and LFF, respectively), oviduct (OF), and uterus (UF) of female dromedary camels. Fluids from small (2-6mm) and large follicles (7-20mm), oviduct and uterus, and blood samples were collected from 19 camels. The results indicated that the concentrations of serum Mg, Fe, and Mn were significantly higher than their follicular fluid, OF, and UF concentrations. Levels of Zn, Fe, Cu, Cr, and Mn were significantly higher in SFF than in LFF. Se and Mo concentrations were higher in LFF. Co concentration was lower in serum than in reproductive tract fluids. Cr concentration was higher in UF and OF than in the serum, SFF, and LFF. High Ca concentration was observed for serum and SFF, followed by LFF. The concentration of Na was about 1.18-fold higher in SFF than in serum, OF, and LFF, and approximately 4.1-fold higher in serum than in UF. K was present in higher concentration in SFF than in serum and LFF; however, its concentration was low in UF and OF. In conclusion, this study shows the concentrations of certain elements in small and large follicular, uterine, and oviductal fluids, which may be low or high depending on their function in the development and growth of follicles. This information can support the development of new media for in vitro oocyte maturation and fertilization of female camels.

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa.

Cells can communicate with other neighboring or distant cells through the secretion of extracellular vesicles (EV), composed of a lipid bilayer and bearing surface molecules that allow them to recognize target cells. In this way, EV induce signaling via different mechanisms, modulating the physiological state of the recipient cell. EV have been identified in both male and female reproductive fluids, however, the possible role of EV isolated from female reproductive fluids has become an emerging field only recently. It is known that ejaculated mammalian spermatozoa need to undergo physiological preparation in the female reproductive tract to fertilize the egg. EV secreted by different regions of the female tract constitute signals that may have a key role in regulating sperm functions. The aims of the present study were isolating EV from different regions of the bovine oviduct and analyzing their interaction and physiological effects on spermatozoa. Here, we report the characterization of bovine oviductal fluid EV from the isthmus and ampulla region and their effect on the induced acrosome reaction and signaling events associated with sperm capacitation. EV induced an increase in sperm protein tyrosine phosphorylation, while cell survival of cryopreserved bovine spermatozoa was maintained. We also show that EV uptake regulates the sperm calcium levels by inducing an immediate increase in the intracellular calcium concentration and sperm priming, after a pre-incubation period, of the progesterone-induced intracellular calcium rise. Our data contribute to understand the role of EV in the communication between the female reproductive tract and the sperm physiology, information that may be used to improve the efficiency of reproductive assisted technologies.

Which Low-Abundance Proteins are Present in the Human Milieu of Gamete/Embryo Maternal Interaction?

The improvement of the embryo culture media is of high relevance due to its influence on successful implantation rates, pregnancy, neonatal outcomes, and potential effects in adult life. The ideal conditions for embryo development are those naturally occurring in the female reproductive tract, i.e., the oviductal and uterine fluids. To shed light on the differences between chemical and natural media, we performed the first comparative study of the low abundance proteins in plasma, uterine, and oviductal fluid collected, simultaneously, from healthy and fertile women that underwent a salpingectomy. The rationale for this design derives from the fact that high-abundant proteins in these fluids are usually those coming from blood serum and frequently mask the detection of low abundant proteins with a potentially significant role in specific processes related to the embryo–maternal interaction. The proteomic analysis by 1D-nano LC ESI-MSMS detected several proteins in higher amounts in oviductal fluid when compared to uterine and plasma samples (RL3, GSTA1, EZRI, DPYSL3, GARS, HSP90A). Such oviductal fluid proteins could be a target to improve fertilization rates and early embryo development if used in the culture media. In conclusion, this study presents a high-throughput analysis of female reproductive tract fluids and contributes to the knowledge of oviductal and uterine secretome.

His Belly, Her Seed: Gender and Medicine in Early Modern Demonic Possession.

This article reassesses the role of gender in early modern demonic possession from a medical perspective. It takes as its starting point the demoniac Richard Mainy, who in 1585 claimed to be suffering from hysteria. Best known for its influence on Shakespeare's King Lear, Mainy's gender-crossing diagnosis should be read in the context of the close historical relationship between hysteria and epilepsy. While medical historians have viewed hysteria as the key possession-related illness, epilepsy was equally important. Both were seen as convulsive illnesses caused by an excess of reproductive fluids. Emphasizing the similarities rather than the differences between male and female sexuality, this shared etiology underpinned medical approaches to demonic possession.

Proteins from male and female reproductive tracts involved in sperm function regulation.

SummarySpermatogenesis is a dynamic process that culminates in the production of mature spermatozoa in the seminiferous tubules of sexually mature animals. Although sperm leaving the testis are fully differentiated, they must further undergo two additional maturation steps before acquiring the capability to fertilize the egg. Such processes take place during the epididymal residency and transport in the seminal fluid during ejaculation and, after delivery into the female reproductive tract, during the journey aiming the encountering the egg in the oviduct. Throughout this trip, spermatozoa are exposed to different reproductive fluids whose molecular compositions regulate the progress towards obtaining a fertilized competent cell. This review summarizes the evidence obtained so far supporting the participation of male and female reproductive tract-derived proteins in the modulation of sperm fertilizing ability and discusses the mechanisms by which such regulation may be accomplished.

Preliminary Study: A New Approach for Improving the Cryopreservation of Mammalian Sperm

Gamete preservation is a necessary and routine procedure practiced in the 21st century in both humans and animals using the cryopreservation technique. However, cryopreservation methods can cause cryoinjury. Therefore, new approaches to help extend the viability of mammalian sperm in vitro are essential. This preliminary study explored the effect of reproductive fluids (RFs) and body fluids (BFs) from two species of desert snails—Sphincterochila zonata and Sphincterochila prophetarum—on mammalian cryopreserved sperm parameters. These desert snails are active only 5% of the year. Spermatogenesis occurs during the aestivation ecophysiological stage when testosterone levels are high, and sperm is preserved in the oviduct until mating during the active ecophysiological stage in winter. RFs from S. zonata and S. prophetarum during the aestivation ecophysiological stage reduced sperm motility to 0%, while sperm viability (SV) was similar to the controls. Moreover, the motility of thawed mouse sperm was 1.34- and 2.02-fold higher (p S. zonata in the active ecophysiological stage than in the control medium after 5- and 30-min incubation. SV was higher in S. zonata RF medium than in control after 30 min incubation. Our results indicate the potential protective effect of desert snail RFs on cryopreserved and thawed mammalian sperm cells. Further study should be conducted to advance the fulfillment of RF potential in reproductive technologies.

The contribution of human sperm proteins to the development and epigenome of the preimplantation embryo.

Knowledge of the proteomic composition of the gametes is essential to understand reproductive functions. Most of the sperm proteins are related to spermatogenesis and sperm function, but less abundant protein groups with potential post-fertilization roles have also been detected. The current data are challenging our understanding of sperm biology and functionality, demanding an integrated analysis of the proteomic and RNA-seq datasets available for spermatozoa, oocytes and early embryos, in order to unravel the impact of the male gamete on the generation of the new individual. The aim of this review is to compile human sperm proteins and to identify and infer their origin and discuss their relevance during oocyte fecundation, preimplantation embryogenesis and epigenetic inheritance. The scientific literature was comprehensively searched for proteomic studies on human sperm, oocytes, embryos, and additional reproductive cells and fluids. Proteins were compiled and functionally classified according to Gene Ontology annotations and the mouse phenotypes integrated into the Mouse Genome Informatics database. High-throughput RNA datasets were used to decipher the origin of embryo proteins. The tissue origin of sperm proteins was inferred on the basis of RNA-seq and protein data available in the Human Protein Atlas database. So far, 6871 proteins have been identified and reported in sperm, 1376 in the oocyte and 1300 in blastocyst. With a deeper analysis of the sperm proteome, 103 proteins with known roles in the processes of fertilization and 93 with roles in early embryo development have been identified. Additionally, 560 sperm proteins have been found to be involved in modulating gene expression by regulation of transcription, DNA methylation, histone post-translational modifications and non-coding RNA biogenesis. Some of these proteins may be critical for gene expression regulation after embryo genome activation, and therefore, may be potentially involved in epigenetic transmission of altered phenotypes. Furthermore, the integrative analysis of the sperm, oocyte and blastocyst proteomes and transcriptomes revealed a set of embryo proteins with an exclusive paternal origin, some of which are crucial for correct embryogenesis and, possibly, for modulation of the offspring phenotype. The analysis of the expression of sperm proteins, at both RNA and protein levels, in tissues not only from the male reproductive tract but also from peripheral organs, has suggested a putative extra-testicular origin for some sperm proteins. These proteins could be imported into sperm from the accessory sex glands and other tissues, most likely through exosomes. These integrative proteome and transcriptome analyses focused on specific groups of proteins, rather than on enriched pathways, identified important sperm proteins which may be involved in early embryogenesis and provided evidence which could support the hypothesis of paternal epigenetic inheritance. The putative extra-testicular origin of some sperm proteins suggests not only the involvement of accessory sex glands in fertilization and epigenetic information transmission, but also that some proteins from additional organs could possibly contribute information to the offspring phenotype. These findings should stimulate further research in the field.

Female nutritional condition affects ovarian fluid quality in guppies.

Male and female gametes are often embedded in fluids that are produced by gonads and other reproductive tissues. Female reproductive fluids, usually called ovarian fluid (OF), which often constitute a relevant volumetric component of the egg mass, are rich in ions, sugars and proteins, and are involved in several functions, from protecting gametes to facilitating fertilization, and often act as mediators of post-mating sexual selection. Despite their applied and evolutionary importance, we know virtually nothing about the costs of female reproductive fluid production. We investigated the effect of nutritional condition on OF quality by experimentally manipulating the diet of two groups of female guppies (Poecilia reticulata) which were maintained for 20 days either on a restricted diet or had ad libitum access to food. In this species, OF enhances sperm swimming longevity and velocity (a predictor of sperm competition success) and mediates post-copulatory inbreeding avoidance. We found that sperm velocity was significantly lower in the OF of diet-restricted females, indicating that OF quality is dependent on female nutritional condition. Our results demonstrate that OF represents a non-trivial component of female reproductive investment and provides a tool to investigate which OF constituents are involved in modulating OF-sperm interactions and fertilization.