Extension of sperm motility leads to increased rates of fertilization and hatching in curimba,Prochilodus lineatus

The objective of this study was to evaluate the effects of six activating solutions on duration of sperm motility, fertilization rate (FR), and hatching rate (HR) of Prochilodus lineatus (Valenciennes, 1837). The activating solutions (SA) used were: SA₀ (199 mOsm kg^?1, pH 8.5), SA₁ (138 mOsm kg^?1, pH 7.5), SA₂ (256 mOsm kg^?1, pH 7.5), SA₃ (131 mOsm kg^?1, pH 10), NaCl (92 mOsm kg^?1, pH 7.5) and distilled water (32 mOsm kg^?1, pH 7.5). SA₁ induced the highest motility, FR and HR, compared with the other activating solutions. The lowest motility was obtained with SA₀, with no fertilization or hatching, whereas motility was zero with SA₂ and SA₃. It is possible to conclude that the solution SA₁ can be used for the activation of gametes during fertilization in induced reproduction of curimba to achieve higher fertilization and hatching rates. Thus, it was found that the osmolality and pH of activating solutions, probably with the participation of dissolved substances therein, are the main factors acting on semen motility after activation.     

Effect of season on bovine seminal plasma proteins in Thailand

Although season has been shown to affect bull sperm quality and fertility in some studies, the effect of season on seminal plasma proteins has not been examined. In the present study, seminal plasma proteins were analysed by Fast Protein Liquid Chromatography (FPLC), to separate the phosphorylcholine-binding proteins and heparin-binding proteins from the other proteins. Semen samples were collected from bulls in three seasons: winter, summer and the rainy season. Sperm quality was analysed by flow cytometry and computer assisted sperm analysis, and further aliquots of semen were used to prepare the seminal plasma for FPLC. Meteorological data were available from a location close to the bull station. There were slight differences in sperm kinematics between seasons, but other parameters of sperm quality were not different. Minor differences in the phosphorylcholine-binding proteins were detected according to season, being lower in summer than in winter or in the rainy season, although there were no changes in the heparin-binding proteins. Temperature, humidity and rainfall differed between winter and the rainy season, but no differences were observed between summer and the rainy season except in the temperature humidity index (THI). However, the THI was above the threshold indicative of heat stress in all seasons, which could explain why few seasonal differences in protein composition were detected in this study. Alternatively, the bulls could have been well-adapted to heat stress. In conclusion, there were only slight differences in bull sperm quality and seminal plasma proteins between seasons during this study.     

哺乳动物及人精子膜离子通道的研究进展

离子的跨膜转动对精子的生理活动起重要的作用。近年,应用膜片钳及人工膜重组等研究通道有关的电生理技术,人们直接观察到哺乳动物及人精子膜上Ｋ、Ｎａ＾＋、Ｃａ＾２＋、Ｃｌ通道的存在。这些结果为揭示精子成熟、获能精卵结合反应等生理过程的某些细节提供了有有的资料,特别是对人精子膜的研究,还为临床应用提供了可能。     

被子植物双受精发现100年：回顾与展望

被子植物双受精发现１００年：回顾与展望＠胡适宜￥北京大学生命科学学院被子植物,双受精,精细胞,卵细胞被子植物双受精发现１００年：回顾与展望胡适宜（北京大学生命科学学院北京１００８７１）关键词被子植物,双受精,精细胞,卵细胞ＣＥＮＴＥＮＡＲＹＯＮＳ．Ｇ．Ｎ...     

Effect of dietary supplementation with a highly pure and concentrated docosahexaenoic acid (DHA) supplement on human sperm function

The aim of this study was to evaluate the possible beneficial effects of diet supplementation with a highly concentrated and purified docosahexaenoic acid (DHA) formula on human sperm function. We performed a prospective, randomized, double blind, placebo-controlled intervention study. One-hundred eighty human semen samples from sixty infertile patients recruited in a private assisted reproduction center were included. All samples were examined according to World Health Organization guidelines. We analyzed macroscopic and microscopic sperm parameters, oxidative stress, apoptosis, lipid peroxidation, mitochondrial membrane potential and DNA fragmentation before and after supplementation with different DHA daily doses (0.5, 1 and 2?g) or placebo for 1 and 3 months. No differences were found in traditional sperm parameters except for progressive sperm motility, with a significant increase after DHA ingestion after the first month with 1 or 2?g doses and after 3 months with 0.5?g of DHA. This effect was more evident in asthenozoospermic patients. No differences were found in any molecular semen parameter except oxidative stress, in which a slight benefit was observed after DHA treatment. In conclusion, this study support previous indications that highlight the importance of DHA supplementation as a means of improving sperm quality in asthenozoospermic men.     

Intramembranal disulfide cross-linking elucidates the super-quaternary structure of mammalian CatSpers

CatSper is a voltage-dependent calcium channel located in the plasma membrane of the sperm flagellum and is responsible for triggering hyperactive motility. A homology model for the transmembrane region was built in which the arrangement of the subunits around the pseudo-four-fold symmetry axis was deduced by the pairing of conserved transmembranal cysteines across mammals. Directly emergent of the predicted quaternary structure is an architecture in which tetramers polymerize through additional, highly conserved cysteines, creating one or more double-rows channels extending the length of the principal piece of the mammalian sperm tail. The few species that are missing these cysteines are eusocial or otherwise monogamous, suggesting that sperm competition is selective for a disulfide-crosslinked macromolecular architecture. The model suggests testable hypotheses for how CatSper channel opening might behave in response to pH, 2-arachidonoylglycerol, and mechanical force. A flippase function is hypothesized, and a source of the concomitant disulfide isomerase activity is found in CatSper-associated proteins β, δ and ε.     

Crystal structure of Brugia malayi venom allergen-like protein-1 (BmVAL-1), a vaccine candidate for lymphatic filariasis

Brugia malayi is a causative agent of lymphatic filariasis, a major tropical disease. The infective L3 parasite stage releases immunomodulatory proteins including the venom allergen-like proteins (VALs), which are members of the SCP/TAPS (Sperm-coating protein/Tpx/antigen 5/pathogenesis related-1/Sc7) superfamily. BmVAL-1 is a major target of host immunity with >90% of infected B. malayi microfilaraemic cases being seropositive for antibodies to BmVAL-1. This study is part of ongoing efforts to characterize the structures and functions of important B. malayi proteins. Recombinant BmVAL-1 was produced using a plant expression system, crystallized and the structure was solved by molecular replacement and refined to 2.1?Å, revealing the characteristic alpha/beta/alpha sandwich topology of eukaryotic SCP/TAPS proteins. The protein has more than 45% loop regions and these flexible loops connect the helices and strands, which are longer than predicted based on other parasite SCP/TAPS protein structures. The large central cavity of BmVAL-1 is a prototypical CRISP cavity with two histidines required to bind divalent cations. The caveolin-binding motif (CBM) that mediates sterol binding in SCP/TAPS proteins is large and open in BmVAL-1 and is N-glycosylated. N-glycosylation of the CBM does not affect the ability of BmVAL-1 to bind sterol in vitro. BmVAL-1 complements the in vivo sterol export phenotype of yeast mutants lacking their endogenous SCP/TAPS proteins. The in vitro sterol-binding affinity of BmVAL-1 is comparable with Pry1, a yeast sterol transporting SCP/TAPS protein. Sterol binding of BmVAL-1 is dependent on divalent cations. BmVAL-1 also has a large open palmitate-binding cavity, which binds palmitate comparably to tablysin-15, a lipid-binding SCP/TAPS protein. The central cavity, CBM and palmitate-binding cavity of BmVAL-1 are interconnected within the monomer with channels that can serve as pathways for water molecules, cations and small molecules.     

Cryopreservation effects on sperm function and fertility in two threatened crane species

The capacity to cryopreserve semen from captive cranes facilitates production of offspring from behaviorally incompatible or geographically separated pairs, and allows for long-term preservation of valuable genetic materials. The present study sought to develop effective cryopreservation protocols for whooping (Grus americana) and white-naped (Grus vipio) cranes, through examining the influences of two permeating (DMA and Me₂SO) and one non-permeating (sucrose) cryoprotectants, as well as vitamin E on post-thaw sperm survival. In Study 1, ejaculates (whooping: n = 10, white-naped: n = 8) were collected and cryopreserved in one of six cryo-diluents (crane extender with: DMA; DMA+0.1M sucrose; Me₂SO; Me₂SO+0.1M sucrose; 0.1M sucrose; 0.2M sucrose) using a two-step cooling method. Frozen samples were thawed and assessed for overall motility, motion characteristics, membrane integrity, morphology, and ability to bind to the inner perivitelline membrane (IPVM). In Study 2, whooping crane ejaculates (n = 17) were frozen in crane extender containing Me₂SO alone or with vitamin E (5 μg/mL or 10 μg/mL). Frozen samples were thawed and assessed as in Study 1, except the binding assay. White-naped crane sperm were more tolerant to cryopreservation than whooping crane (15% vs 6% post-thawed motility). In both species, sperm cryopreserved in medium containing Me₂SO alone displayed higher post thaw survival and ability to bind to IPVM than the other cryodiluent treatments. Vitamin E supplementation exerted no benefits to post thaw motility or membrane integrity. The findings demonstrated that there was species specificity in the susceptibility to cryopreservation. Nevertheless, Me₂SO was a preferred cryoprotectant for sperm from both whooping and white-naped cranes.     

Attenuation of cryopreservation-induced oxidative stress by antioxidant: Impact of Coenzyme Q10 on the quality of post-thawed buck spermatozoa

The aim of this study was to determine the quality of post-thawed buck spermatozoa by attenuation of cryopreservation-induced oxidative stress using CoQ₁₀, a lipophilic antioxidant. Ejaculates at every sampling period were collected from four Mahabadi bucks, pooled and diluted in soybean lecithin-based extenders containing 0 (negative control, NC), 0.5 (CQ0.05), 1 (CQ1), and 1.5 (CQ1.5) μM CoQ₁₀ and 0.9% (v/v) DMSO (positive control, PC). The diluted semen was gradually cooled to 4 °C, then frozen and stored in liquid nitrogen. After thawing, total motility although was significantly higher in CQ1 (53.40 ± 1.83) than control groups (43.60 ± 1.83% and 42.20 ± 1.83%; P < 0.05), but this parameter did not differ between CQ1 and CQ1.5. Sperm viability was significantly higher in CQ1 (54.20 ± 2.03%) than that of control and CQ0.5. The CQ1 and CQ1.5 led to significantly higher the plasma membrane functionality compared to control groups. Sperm abnormality was significantly lower in CQ1 than that of NC. The results also showed that MDA level was significantly lower in CQ1 and CQ1.5 compared with control and CQ0.5. The CQ1 (59.43 ± 3.93%) was significantly increased mitochondrial activity compared to control groups. Although a greater value for %DFI was found in NC (10.24 ± 0.48%) and PC (9.77 ± 0.48%) groups compared to others, it was lower in CQ1 group (4.26 ± 0.48%). In conclusion, based on our research results, 1 μM CoQ₁₀ could protect buck spermatozoa from cryoinjury.