Sperm storage tubules (SST) are central to fertility in birds, yet their structure and anatomical location have not been well characterized in ducks. This gap has limited direct study of the mechanisms that drive variation in fertility persistence among females used in intergeneric crosses. Here, we labeled duck spermatozoa with Hoechst 33342 and traced their distribution in the oviduct after artificial insemination. Staining conditions were optimized across temperature, dye concentration, and incubation duration. The selected protocol (40 µg/mL, 40 min, 37°C) produced strong fluorescence without compromising motility. Labeled spermatozoa were inseminated into Wujie White duck females, a line bred for prolonged fertility persistence in crosses with Muscovy drakes. Oviducts collected 24 hr after the second insemination were examined by fluorescence microscopy, revealing clusters of labeled spermatozoa within the mucosal folds of the utero-vaginal junction (UVJ). Hematoxylin and eosin staining confirmed that SST were present exclusively at the UVJ of the lower oviduct. Unlike conventional histology, the present approach combines fluorescent sperm labeling, artificial insemination, and direct examination of isolated UVJ villi. This permits concurrent evaluation of SST morphology and intraluminal sperm distribution at the individual level. It can also serve as a practical tool for future comparative studies of sperm retention in females with contrasting fertility persistence. Histological examination of five additional non-inseminated females further verified the consistent presence of SST in UVJ villi across individuals. To our knowledge, this study represents the first direct visualization and localization of intact SST in the duck oviduct, and defines an anatomical reference and targeted sampling strategy for reproductive research in waterfowl.
This review provides an up-to-date overview of the roles of the oviduct during the periconception period and underlying mechanisms. The functions of the oviduct before, during, and after fertilization are highlighted, with special focus on the effects of epithelial cell contact and luminal secretions on sperm selection mechanisms and acquisition of fertilization ability. The current knowledge on how the oviduct contributes to support fertilization and embryo development via the overall physical milieu (oxygen tension, fluid current, ciliated epithelial cells) and the role of its secretions is also provided. Altogether, the review underlines the unique role of the oviduct during gamete selection and early embryo development, which so far has not been completely possible to mirror when assisted reproductive technologies (ART) are used. Unveiling the most important functional components of oviductal secretions that contribute to better sperm selection, and boost sperm fertilizing ability and early embryo development, can indeed be useful to improve the outcomes of current in vitro systems used in ART.
The purpose of this study was to investigate the effects of antioxidant compounds derived from synthetic sources (a combination of 200 mg vitamin C/kg, 150 mg vitamin E/kg, 0.30 mg Se yeast/kg, and 150 mg carnitine/kg) and phytogenic sources (a 1 % combination of clove, green tea pomace, and Vietnamese coriander) in breeder hens under heat stress (HS) on reproductive performance, egg quality, and offspring growth performance, and their antioxidant capacity. One hundred breeder hens were randomly divided into four treatments to receive either a basal diet or a basal diet supplemented with synthetic or phytogenic antioxidants under thermoneutral (TN) (23 +/- 1 degrees C) and chronic HS (36 +/- 1 degrees C for 4 h/day) conditions: T1: TN + basal diet, T2: HS + basal diet, T3: HS + synthetic antioxidants, and T4: HS + phytogenic antioxidants. The results showed that either synthetic or phytogenic antioxidants can improve the Haugh unit and maintain ovary weight, and large and small yellow follicles (P < 0.05). In addition, phytogenic antioxidants were also found to enhance egg yolk color. Although the body weight of offspring from either antioxidant source was initially low on day 1, it subsequently increased until reaching levels comparable to those in the TN and HS without supplementation by 3 weeks of age (P > 0.05). Either antioxidant source can improve free radical scavenging, reduce lipid peroxidation, upregulate the relative expression of SOD, CAT, and GSH-Px mRNA, and downregulate HSP90 in offspring liver (P < 0.05). This study highlights the beneficial effects of combining either synthetic or phytogenic antioxidants to mitigate the negative impact of HS on breeder hens and the maternal antioxidant properties transmitted to their offspring.
In brief:The molecular interactions between spermatozoa and the oviduct epithelium that are involved in capacitation and sperm availability during fertilization still remain largely unknown in many species. This study provides comprehensive proteomes of the apical and basal part of the bovine oviduct epithelium (isthmus and ampulla, pre- and post-ovulatory) and presents new protein candidates for sperm binding in the bovine functional sperm reservoir. Abstract:The proximal region of the oviduct (isthmus) serves as a sperm reservoir in many mammalian species. This reservoir is composed of epithelial cells that bind sperm primarily to their apical cilia and ensure the survival of high-quality sperm until ovulation. The species-specific molecular interactions between spermatozoa and the oviduct epithelium, however, are still poorly understood. The aim of this study is to identify new candidates for sperm-interacting proteins in the bovine reservoir using laser capture proteomics. Ipsilateral oviducts were collected from cyclic cows at pre- and post-ovulatory stages. Isthmus and ampulla tissue samples were fixed, paraffin-embedded, sectioned and subjected to laser microdissection to establish pools of microsamples separating the apical part of the epithelium, containing potential sperm-interacting proteins, from the basal part, containing proteins not currently located or secreted on the cell surface. Proteins were analyzed by nanoLC-MS/MS, and differentially abundant proteins (DAPs) were identified using t-tests. A total of 505 and 512 proteins could be identified in apical and basal microsamples, respectively. Only 8/141 regional and 1/79 cycle stage-related DAPs were shared between both epithelium parts, indicating a selective regulation of protein expression. Of the apical proteins, 19 were predicted to be candidates for sperm interactions, including annexins (ANXA) 1, 2, 5 and 8; oviduct glycoprotein 1 (OVGP1); voltage-dependent anion-selective channel proteins (VDAC) 1 and 2; apolipoprotein A1 (APOA1). This work provides the first proteomic characterization of microdissected cellular compartments of the bovine oviduct epithelium and presents new candidates for improving sperm quality in assisted reproductive technologies.
The sperm reservoir in the isthmus, the distal part of the oviduct, plays important roles in sperm survival and acquisition of sperm fertilization competence. Here, we describe an in vitro model of bovine oviduct spheroids, easy to culture and normalize, and allowing robust quantification by confocal microscopy of sperm binding capacity to isthmic epithelial cells. This assay may be used to compare sperm binding capacity between males and ejaculates, evaluate sperm membrane integrity and correlation with male fertility, and conduct studies on sperm-oviduct interactions. The method may be implemented in other species.
Abstract Background In vitro embryo production is increasingly used for genetic improvement in cattle but bypasses the oviduct environment and exposes the embryos to oxidative stress with deleterious effects on further development. Here we aimed to examine the effect of oviduct epithelial spheroids (OES) on embryo development and quality in terms of morphology and gene expression during two co-culture times (4 days: up to embryonic genome activation at 8–16 cell stage vs. 7 days: up to blastocyst stage) and under two oxygen levels (5% vs. 20%). Methods Bovine presumptive zygotes produced by in vitro fertilization (day 0) using in-vitro matured oocytes were cultured in droplets of synthetic oviductal fluid (SOF) medium with or without (controls) OES for 4 or 7 days under 5% or 20% oxygen (4 treated and 2 control groups). Cleavage rates were evaluated on day 2 and blastocyst rates on days 7–8. Expanded blastocysts on days 7–8 were evaluated for total cell numbers and gene expression analysis by RNA-sequencing. Results Under 20% oxygen, blastocyst rates and total cell numbers were significantly higher in the presence of OES for 4 and 7 days compared to controls (P < 0.05), with no difference according to the co-culture time. Under 5% oxygen, the presence of OES did not affect blastocyst rates but increased the number of cells per blastocyst after 7 days of co-culture (P < 0.05). Both oxygen level and OES co-culture had a significant impact on the embryonic transcriptome. The highest number of differentially expressed genes (DEGs) was identified after 7 days of co-culture under 20% oxygen. DEGs were involved in a wide range of functions, including lipid metabolism, membrane organization, response to external signals, early embryo development, and transport of small molecules among the most significantly impacted. Conclusion OES had beneficial effects on embryo development and quality under both 5% and 20% oxygen, mitigating oxidative stress. Stronger effects on embryo quality and transcriptome were obtained after 7 than 4 days of co-culture. This study shows the impact of OES on embryo development and reveals potential molecular targets of OES-embryo dialog involved in response to stress and early embryonic development.
Currently, glycerol is the most effective cryoprotectant when combined with straw packaging for preserving chicken sperm. Glycerol, however, has toxic effects on sperm cells, which can reduce fertility when present in inseminated semen. Historically, the serial dilution (SD) method was developed to eliminate glycerol and mitigate its adverse effects. We have recently developed a new method for removing glycerol called sucrose-Percoll (SP), that can be performed at either 4°C (4°C-SP) or 20°C (20°C-SP). This SP protocol has been found to be simpler and faster to improve fertility compared to the traditional SD method. Nevertheless, the reasons for such effectiveness differences between glycerol removal procedures remained unclear and required more comprehensive understandings for future protocol developments. Here, we examined the effects of SP and SD protocols on the fertility duration. We also investigated the potential causes of varying effects of these methods by analyzing sperm quality parameters and sperm storage in the hen's reproductive tract. The fertility was significantly higher in 4°C-SP than 20°C-SP during the first 6 d after insemination, and also higher than sperm processed using SD. No difference was observed between 20°C-SP and SD between 7 and 13 d. However, a 2.7-time higher fertility was shown with 4°C-SP. In addition, the SP method demonstrated a 2-fold greater ability to remove glycerol than the SD method. Sperm centrifuged at 4°C-SP exhibited higher sperm storage compared to 20°C-SP and were higher than sperm treated with SD. Overall, our findings revealed that the differences in efficiencies between SP and SD methods were not related to in vitro sperm quality but resulted from a higher ability to remove glycerol, a higher storage capacity in the female reproductive tract, and a longer fertility ability. Since no impacts were observed in sperm cellular characteristics, further experiments are necessary to investigate the influences of glycerol removal treatments at the molecular level.
Most in vitro models of oviduct epithelial cells (OEC) used thus far to gain insights into embryo-maternal communication induce cell dedifferentiation or are technically challenging. Moreover, although the presence of developing embryos has been shown to alter gene expression in OEC, the effect of embryos on OEC physiology remains largely unknown. Here, we propose a model based on bovine oviduct epithelial spheroids (OES) with specific shape and diameter (100-200 mu m) criteria. The aims of this study were to i) determine the appropriate culture conditions of bovine OES cultured in suspension by evaluating their morphology, total cell number, viability, and activity of ciliated cells; ii) monitor gene expression in OES at the time of their formation (day 0) and over the 10 days of culture; and iii) test whether the vicinity of developing embryos affects OES quality criteria. On day 10, the proportions of vesicle-shaped OES (V-OES) were higher in M199/500 (500 mu l of HEPESbuffered TCM-199) and synthetic oviduct fluid (SOF)/25 (25-mu L droplet of SOF medium under mineral oil) than in M199/25 (25-mu L droplet of M199 under mineral oil). The proportion of viable cells in V-OES was not affected by culture conditions and remained high (>80%) through day 10. The total number of cells per V-OES decreased over time except in SOF/25, while the proportions of ciliated cells increased over time in M199/500 but decreased in M199/25 and SOF/25. The movement amplitude of OES in suspension decreased over time under all culture conditions. Moreover, the gene expression of ANXA1, ESR1, HSPA8, and HSPA1A in OES remained stable during culture, while that of PGR and OVGP1 decreased from day 0 to day 10. Last, the co-culture of developing embryos with OES in SOF/25 increased the rates of blastocysts on days 7 and 8 compared to embryos cultured alone, and increased the proportion of V-OES compared to OES cultured alone. In conclusion, M199/ 500 and SOF/25 provided the optimal conditions for the long-time culture of OES. The supporting effect of OES on embryo development and of developing embryos on OES morphology was evidenced for the first time. Altogether, these results point OES as an easy-to-use, standardizable, and physiological model to study embryo-maternal interactions in cattle.
The objective of the present study was to transpose sperm freezing methodology from domestic goat to the Tadjik markhor (Capra falconeri heptneri) and to address the feasibility to develop IVP and artificial insemination using such frozen semen. Semen of different adult markhor males were successfully recovered by electro-ejaculation and were then frozen using caprine methodology. Frozen semen showed good survival rates at thawing and good fertility rates were assessed in heterologous in vitro fertilization system with goat oocytes. LOPU/IVF was applied for Tadjik markhor females allowing the first successful blastocyst production in vitro. In an applied program, we also transposed successfully intrauterine AI method with frozen/thawed semen to the Tadjik markhor.
The elimination of ejaculates and males with low fertility despite good sperm motility and morphology is crucial to maintain high pregnancy rates after artificial insemination (AI) in farm animals. The ability of sperm to survive in the female tract is particularly crucial in pigs due to the large variation in the timing between AI and ovulation and the high number of oocytes to fertilise. The objective of this study was to characterise a new in vitro model of oviduct sperm reservoir using porcine oviduct epithelial spheroids (OES) and to assess the variability in sperm binding to OES among gilts, boars and their ejaculates. Isthmic mucosa fragments were collected from gilt oviducts at a slaughterhouse, and after 48 h of culture, the OES that had spontaneously formed were sorted according to their vesicle shape and size (150-200 mu m in diameter) for characterisation and sperm binding assays. The OES contained viable, cytokeratin-positive and vimentin-negative cells, of which 36.4 +/- 2.0% were multiciliated. The average proportion of multiciliated cells per OES did not change among culture replicates. After co-incubation with boar fresh semen, only sperm of normal morphology were found to bind, by their head, to cilia of OES. The density of sperm bound to the OES surface increased linearly with sperm concentration. The bound sperm density on OES was used to assess the binding capacity of fresh ejaculates collected from Pietrain boars. For a given ejaculate, the bound sperm density did not vary among pools of OES female donors. The analysis of five successive ejaculates from nine boars indicated significant differences in bound sperm densities on the OES among individual boars and their ejaculates (P < 0.01). There was no correlation between the sperm bound density and sperm parameters measured by computer-assisted sperm analysis or the initial dilution of the ejaculate. In conclusion, the OES characterised in this study offered physiological conditions to study sperm binding to the isthmic reservoir and evidenced that sperm from different ejaculates and different boars vary in their ability to bind to these oviduct spheroids despite homogeneous motility and morphology.
This study aimed to investigate the efficacy of a synthetic source (a combination of vitamin E, vitamin C, selenium, and L-carnitine) and phytogenic sources (a combination of clove, green tea pomace, and Vietnamese coriander) in overcoming heat stress (HS) damage in female breeder hens on production, blood chemistry, sperm survival in the oviduct, antioxidant properties, gene expression, and quality of offspring. One hundred SUT female breeder hens were housed in individual cages and divided into 4 treatment groups: T1) basal diets in the thermoneutral (TN) zone; T2) basal diets under HS; 3) basal diets with synthetic antioxidants under HS; and T4) basal diets with phytochemical antioxidants under HS. The result revealed that HS condition had a negative effect on reducing final body weight, egg weight, and 1-day-old chick weight while increasing water intake and FCR and altered blood chemicals in breeder hens compared to TN breeder hens (P < 0.05). However, either synthetic or phytogenic antioxidants resulted in increased egg production and hatchability, while decreasing the number of late stages of embryo death during the incubation (P < 0.05). Furthermore, the synthetic antioxidants also improved the uniformity of chicks and reduced late-stage embryo death compared with phytogenic antioxidants (P < 0.05). HS breeder hens fed with either of the antioxidant sources exhibited higher antioxidant capacity in terms of DPPH and ABTS radical scavenging (in yolk, liver, and breast meat) and FRAP radical scavenging (in yolk and liver) and lower liver malondialdehyde than HS breeder hens fed with the control diet (P < 0.05). Additionally, the gene expression of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) in the liver was upregulated, whereas the expression of proinflammatory cytokines (nuclear factor-κB) and heat shock proteins (HSP70 and HSP90) was downregulated in breeder hens that received both antioxidant sources (P < 0.05). Future investigations should focus on the potential for combinations of synthetic and phytogenic antioxidants in diets for HS breeder hens.
Excessive free radicals in human and animal bodies can cause oxidative stress (OS) which damages cells and tissues. Plant materials with high antioxidant potential would resolve the OS problem. Thus, this study proposed to investigate the total phenolic (TPC) and flavonoid contents (TFC), antioxidant capacities and cytotoxicity in 17 edible plant materials from herbs, fruits, vegetables and plant by-products available in Southeast Asia for future use in the food or feed industry. Among 17 plant materials, Syzygium aromaticum (clove), Camellia sinensi (green tea pomace) from the beverage industry and Persicaria odorata (Vietnamese coriander) showed a prominent amount of TPC and TFC. These three plants and their combination (1:1:1 ratio, v:v:v) also possessed a remarkable antioxidant function in terms of DPPH, ABTS and FRAP, as well as showing a strong ROS inhibition through HepG2 cells. The cytotoxicity test of the crude extract of clove, green tea pomace and Vietnamese coriander, or their combination can be used between 0.032 and 0.255, 0.011 to 0.088, 0.022 to 0.178 and 0.021 to 0.346 mg/mL, respectively, without impeding cell viability. A combined mixture of clove, green tea pomace and Vietnamese coriander revealed the synergistic properties of antioxidants and cell safety. This indicates that there is a potential use of various antioxidant bioactive compounds in plant materials tested for use as phytogenic antioxidant additives.
A concentration of 11% of glycerol is the standard one for sperm cryopreservation in chickens, however, the presence of just 2% glycerol already causes severe fertility reduction, suggesting the necessity of removing glycerol before artificial insemination (AI). The major approach developed for this purpose is serial dilution followed by centrifugation (SDC), which demands special equipment (such as a refrigerate room) to maintain post-thaw semen at 4 °C, besides being time consuming. Therefore, we attempted to develop a simple method to remove glycerol from chicken frozen-thawed semen based on a colloidal gel, Percoll, which is ordinarily used to select motile and viable sperm in mammals as well as in fresh chicken semen. In this study, we used a Percoll based glycerol removal solution (GRS) containing sucrose to avoid frozen-thawed sperm suffering from osmotic stress. Subsequently, several conditions including GRS compositions (GRS A, B, C and D) and centrifugation temperatures (4 and 20 °C) were compared by their influence on sperm in vitro parameters. Afterwards, GRS A and D were selected for fertility evaluation, compared to conventional SDC method. Our results showed that the fertility with GRS A at both 4 and 20 °C were higher than GRS D (p < 0.05) and similar or even superior to the fertility obtained with SDC method. Altogether, our novel GRS protocol is a valuable method for chicken sperm cryobanking policy, supported by its notable results of fertility as well as saving 44% of time, with a simple equipment at flexible operation temperatures of 4 or 20 °C.
The objective of this study was to inves-tigate the effects of modified dietary fiber from cassava pulp (M-DFCP) supplementation in broiler diets on cecal microbial populations, short-chain fatty acids (SCFAs), ammonia production, and immune responses. A total of 336, one-day-old male broiler chicks (Ross 308) were distributed over 4 dietary treat-ments in 7 replicate pens (n = 12 chicks) using a completely randomized design. Chicks were fed the control diet and 3 levels of M-DFCP (0.5, 1.0, and 1.5%) for an experimental duration of 42 d. The M-DFCP contained total dietary fiber (TDF), soluble dietary fiber (SDF), insoluble dietary fiber (IDF), cello-oligosaccharides (COS), and xylo-oligosacchar-ides (XOS) of approximately 280.70, 22.20, 258.50, 23.93, and 157.55 g/kg, respectively. The 1.0 and 1.5% M-DFCP supplementation diets showed positive effects on stimulating the growth of Lactobacillus spp. and Bifidobacterium spp., enhancing SCFAs (acetic, propionic, butyric acid, and branched SCFAs) and lac-tic acid concentrations during growing periods. Broilers fed 1.0 and 1.5% M-DFCP also exhibited a sig-nificant increase in caecal Lactobacillus spp. and lactic acid concentrations during the finisher period as well. In addition, M-DFCP also reduced cecal digesta and excreta ammonia production in broilers over both peri-ods (0-21 and 22-42 d of age). However, M-DFCP did not exhibit any effect on total serum immunoglob-ulin (Ig) or lysozyme activity. In conclusion, this study shows that M-DFCP can be used as a dietary fiber source in broiler diets, with a recommended level of approximately 1.0%.
IntroductionDue to their capacity to release growth factors and cytokines, co-culture using mesenchymal stem cells has been considered a good alternative to promoting the maturation of the oocytes and the embryo’s development quality in vitro in different mammalian species. In this regard, we investigated the effect of feline Wharton’s jelly MSCs as feeders layer in oocyte maturation—consequently, the development of resulting embryos in co-culture.MethodsOocytes with dark cytoplasm and a few layers of cumulus cells were collected and subjected to in vitro maturation and embryo culture using commercial media with and without MSCs addition. The oocytes’ nuclear maturation and the degree of cumulus expansion in different groups were assessed after 24 h; the development of the embryo was evaluated every 12 h until day eight.ResultsAlthough MSCs increased the proportion of cumulus cells oocytes exhibiting cumulus expansion, there were no significant differences in the percentage of matured oocytes (metaphase II) among the groups (p > 0.05). However, the embryo development differs significantly, with a higher cleavage, morula, and blastocyst percentage in oocytes matured with MSC co-culture conditions than in commercial media alone (p < 0.05). Also, we observed higher morula and blastocyst rates in the embryos co-cultured with MSCs during the in vitro culture (p > 0.05).ConclusionBased on our results, the co-culture with MSCs during the oocyte maturation resulted in better embryo development, as well as the MSCs addition during embryo culture returned an increased number of morula and blastocysts. Further research is needed to fully understand and optimize the use of MSCs in oocyte maturation and embryo development.
Glycerol is a cryoprotectant used widely for the cryopreservation of animal sperm, but it is linked to a decrease in fertility. The mechanism underlying the negative effects of glycerol remains unclear. Therefore, in this study, we aimed to gain a better understanding by using the chicken model. First, we investigated the impact of increasing the concentration of glycerol during insemination on hen fertility. Our findings revealed that 2% glycerol resulted in partial infertility, while 6% glycerol led to complete infertility. Subsequently, we examined the ability of sperm to colonize sperm storage tubules (SST) during in vivo insemination and in vitro incubation. The sperm used in the experiment were stained with Hoechst and contained 0, 2, or 6% glycerol. Furthermore, we conducted perivitelline membrane lysis tests and investigated sperm motility, mitochondrial function, ATP concentration, membrane integrity, and apoptosis after 60 min of incubation with different glycerol concentrations (0%, 1%, 2%, 6%, and 11%) at two temperatures to simulate pre-freezing (4 °C) and post-insemination (41 °C) conditions. Whereas 2% glycerol significantly reduced 50% of sperm containing SST, 6% glycerol completely inhibited SST colonization in vivo. On the other hand, in vitro incubation of sperm with SST revealed no effect of 2% glycerol, and 6% glycerol showed only a 17% reduction in sperm-filled SST. Moreover, glycerol reduced sperm-egg penetration rates and also affected sperm motility, bioenergetic metabolism, and cell death at 4 °C. These effects were observed when the concentration of glycerol exceeded 6%. Furthermore, at 41 °C, glycerol caused even greater damage, particularly in terms of reducing sperm motility. These data altogether reveal important effects of glycerol on sperm biology, sperm migration, SST colonization, and oocyte penetration. This suggests that glycerol plays a role in reducing fertility and presents opportunities for improving sperm cryopreservation.
Background Spermatozoa interact with oviduct secretions before fertilization in vivo but the molecular players of this dialog and underlying dynamics remain largely unknown. Our objectives were to identify an exhaustive list of sperm-interacting proteins (SIPs) in the bovine oviduct fluid and to evaluate the impact of the oviduct anatomical region (isthmus vs. ampulla) and time relative to ovulation (pre-ovulatory vs. post-ovulatory) on SIPs number and abundance. Methods Pools of oviduct fluid (OF) from the pre-ovulatory ampulla, pre-ovulatory isthmus, post-ovulatory ampulla, and post-ovulatory isthmus in the side of ovulation were collected from the slaughterhouse. Frozen-thawed bull sperm were incubated with OF or phosphate-buffered saline (control) for 60 min at 38.5 °C. After protein extraction and digestion, sperm and OF samples were analyzed by nanoLC-MS/MS and label-free protein quantification. Results A quantitative comparison between proteins identified in sperm and OF samples (2333 and 2471 proteins, respectively) allowed for the identification of 245 SIPs. The highest number (187) were found in the pre-ovulatory isthmus, i.e., time and place of the sperm reservoir. In total, 41 SIPs (17%) were differentially abundant between stages in a given region or between regions at a given stage and 76 SIPs (31%) were identified in only one region × stage condition. Functional analysis of SIPs predicted roles in cell response to stress, regulation of cell motility, fertilization, and early embryo development. Conclusion This study provides a comprehensive list of SIPs in the bovine oviduct and evidences dynamic spatio-temporal changes in sperm-oviduct interactions around ovulation time. Moreover, these data provide protein candidates to improve sperm conservation and in vitro fertilization media.