Reproduction in Domestic Animals最新文献

Lipids in cumulus-oocyte complexes are important actors in molecular signalling pathways and are influenced by maturation conditions. Acetyl-L-carnitine (ALC) is a carrier involved in fatty acid transport and is a promoter of β-oxidation. Although the embryonic development potential of oocytes can be improved when ALC is added to the maturation medium, the effects of ALC on the lipid content and composition of oocytes and cumulus cells remain unknown. In this study, the effect of ALC supplementation on the lipid profiles of buffalo oocytes and cumulus cells after in vitro maturation was evaluated using positive-ion matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Orthogonal projections to latent structures discriminant analysis was used to analyse the data. The lipid profiles of oocytes and cumulus cells differed following ALC treatment. Four lipids were significantly different in oocytes and eight in granulosa cells. ALC treatment significantly reduced the cellular content of these lipids, with the exception of phosphatidylcholine [PC(36:3) + H] + in oocytes and triacylglycerol [TAG(58:8) + NH4] + in granulosa cells. Further staining experiments confirmed that ALC treatment reduced the total lipid content in oocytes. Therefore, ALC treatment alters lipid metabolism in oocytes and cumulus cells during their maturation and accelerates lipid metabolism in oocytes. The findings of this study demonstrate that ALC enhances the rate of oocyte maturation by altering lipid metabolism in oocytes, providing both a clear direction for further research into regulatory mechanisms as well as a foundation for further improving oocyte maturation rates.

Urine samples were systematically collected from inseminated Murrah buffaloes (Bubalus bubalis) on days 0, 7, 14, 21 and 28 (with day 0 representing the day of artificial insemination). Following confirmation of pregnancy via trans rectal palpation 45 days of insemination, the animals were categorised into pregnant and non-pregnant groups (n = 10 each). The urine samples on 0, 7, 14, 21 and 28 days of pregnant and one sample from non-pregnant preferably collected on 28th day was used for SDS-PAGE after diafiltration. We focused on urine samples obtained on 28th day of insemination as it could contain high fetuin-A levels associated with pregnancy. Each of the two urine samples from both the pregnant and non-pregnant groups underwent purification through wheat germ agglutinin (WGA) affinity separation. The resulting fraction/elute was subsequently employed for the determination and validation of fetuin-A through 1D SDS-PAGE, western blotting and MALDI-TOF analysis. The samples were used for measuring the concentration of fetuin-A by ELISA. Fetuin-A was significantly higher (1020.50 ± 19.75 mg/L) in pregnant Murrah buffalo urine than their non-pregnant counterparts (86.5 ± 6.33 mg/L), with a p < 0.05. On SDS-PAGE analysis, the WGA fractions revealed distinct protein bands, notably between ~54 to 70 kDa and slightly above ~91 kDa, in pregnant Murrah buffaloes compared with their non-pregnant counterparts. Following confirmation of fetuin-A at ~58 kDa through western blotting, the corresponding protein band was excised from the SDS gel and subjected to mass spectrometry for MALDI-TOF analysis. Peptide mass fingerprinting, coupled with protein BLAST, confirmed the upregulated protein as Alpha2-HS glycoprotein, also known as fetuin-A. These findings are crucial for elucidating the presence of fetuin-A in urine samples from pregnant Murrah buffaloes, offering valuable insights into their physiological status.

Spermatogenesis is a highly complex and tightly regulated cellular differentiation process closely related to the productive performance of male livestock. We do not yet have a clear understanding of the spermatogenesis mechanism of buffalo. In this study, spermatogonia, spermatocytes and spermatids were analysed by flow cytometry. Quantitative proteomic and phosphoproteomic studies were performed on different spermatogenic cells using tandem mass tagging technology and liquid chromatography-tandem mass spectrometry. A total of 219 differentially expressed proteins (involved in focal adhesions and actin cytoskeleton pathways) and 71 phosphoproteins (involved in RNA transport and adhesion junction pathways) were obtained. Through trend analysis, a dynamic profile of protein expression was obtained, enriched to the main biological processes at different stages of spermatogenesis. By immunohistochemical localisation analysis, it was found that MACROH2A2, TOP2A, LMNA, LMNA (pS392), VIM and VIM (pS56) had specific localisation in testis cells. Network analysis of kinase-substrate phosphorylation sites showed that AKT1 is the most active kinase, LMNA is regulated by most kinases and AKT1 can catalyse the phosphorylation of LMNA. This study provides a reference for studying the molecular mechanism of buffalo spermatogenesis and helps clarify the regulatory mechanism of protein translation and post-translational modification during mammalian spermatogenesis.

microRNA (miRNA) is a class of small non-coding RNA molecules that are widely expressed in organisms and play an important role in the regulation of gene expression at the post-transcriptional level. In recent years, researchers have begun to explore its effects on the development of domestic animals and have begun to think about its potential role in modern molecular breeding. Increasing evidence shows that miRNA play a central role in the regulation of pig fertility, pork product quality and disease resistance. Understanding the physiological mechanism of miRNA will be able to better guide future breeding work. In this paper, we will review the research progress of the function and mechanism of miRNA in combination with the above economic characteristics of pigs. The reported miRNA and their target genes were sorted out to evaluate their potential role in improving economic traits such as pig fertility, meat quality and disease resistance, to provide a reference for modern pig molecular breeding.

Bull fertility is a multi-factorial trait and is affected by many factors, such as nutrition, genetics, and epigenetics. Superior quality male germplasm with high genetic merit helps to improve the livestock production trait. To achieve the target of livestock production, the availability of superior male germplasm is a great concern. In developing countries, there is a gap between the highly fertile frozen semen doses produced and the highly fertile frozen semen doses required. Improving the quality of existing low-fertile semen from high genetic merit bulls seems to play a promising role in filling this gap. Seminal exosomes are extracellular vesicles secreted by the epithelial cells of the testis, epididymis, and accessory sex glands such as the prostate gland. They contain a cargo of bioactive molecules such as proteins, nucleic acids and various metabolites. These molecules are transferred to the spermatozoa during its maturation and help in sperm capacitation, maturation, acrosome reaction, and fertilisation. Studies reveal that seminal exosomes help to improve the sperm functionality of low-quality sperm. Identification of the molecular profile of exosomes of bulls with proven fertility and their addition in an extender containing low-fertile semen may help to ameliorate the sperm quality of low-fertile semen, which may eventually aid in generating quantities of highly fertile ejaculates.

Current study was aimed to assess the β-cyclodextrin (βCD) and methyl-β-cyclodextrin (MβCD) on delivery of cholesterol, and substitution of fructose and glycerol with trehalose on the ram semen cryosurvival. Samples were collected, diluted with Tris-citric acid-LDL extender, pooled, and used. In experiment I, βCD and MβCD carriers were used and compared to deliver the cholesterol (at 0, 0.5, 1, 1.5, 2, and 4 mg/mL). In the experiment II, trehalose (0, 7, 14, 21, and 28 mM) was substituted with fructose (28, 21, 14, 7, 0 mM, respectively). In the experiment III, the best cholesterol/carrier dose groups from the first experiment, were selected to be evaluated with the fructose/trehalose (14/14 mM) combination compared to fructose (28 mM) alone. The concentration of glycerol in the above-mentioned experiments was set at 4.5%. In the experiment IV, the effect of lowering glycerol (4% vs. 4.5%) was assessed using selected cholesterol/carrier groups. Kinematics, chromatin integrity, ATP contents, malondialdehyde amounts and viability were evaluated. Cholesterol (especially at 1.5 and 2 mg/mL) improved the kinematics and ATP levels using both carriers. The optimised amounts of trehalose (14 mM)/fructose(14 mM) reduced peroxidation and DNA fragmentation levels. Co-administration of optimised levels of cholesterol with trehalose/fructose did not show extra beneficial effects compared to each of them. Trehalose could not protect the spermatozoa at lower amounts of glycerol (4% vs. 4.5%). In conclusion, either the optimised levels of cholesterol (using βCD or MβCD carriers) or substitution of half of the fructose with the trehalose alone could lead to improvement in quality of frozen/thawed ram semen.

This article describes the carbohydrate composition of early and mature placentas from bitches, detected by lectin histochemistry. Formalin-fixed placental samples from 11 mixed-breed bitches have been assigned to the 'early' or the 'mature' placenta group, processed by the routine histological technique and labelled with a panel of 14 biotinylated lectins. The glycan distribution was almost completely preserved over pregnancy. Cytotrophoblast cells expressed high-mannose N-linked residues and N-acetyl-D-glucosamine, whereas mannose N-linked residues, N-acetyl-D-glucosamine and β- and α-D-galactose/N-acetyl-D-galactosamine have been found in the syncytiotrophoblast. The maternal and foetal endothelial binding pattern was enriched in β-D-galactose, α-D-mannose in non-bisected bi/tri-antennary N-linked complex and Galα1,3Gal-Galα1,4Gal. Both deep and superficial glands showed a great variety of glycoconjugates, such as D-mannose, D-glucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, galactose, N-acetylneuraminic acid and fucose. Most results in this study were consistent with those previously reported in canine and feline mature placentas; here, early placentas have been analysed and the lectin binding pattern of mature placentas has been further described. These studies on canine placentas contribute to detecting and understanding glycome changes in pathophysiological conditions.

As the hybrid between cattle and yak, cattleyak is a typical male sterile mammal, and the underlying mechanism for its spermatogenic arrest is still unclear. In this study, the coding region of cattleyak TAF4B gene was cloned by RT-PCR and analysed by bioinformatics. To investigate the effects of TAF4B on cellular proliferation and differentiation, an expression vector was generated and introduced into undifferentiated spermatogonia (UDSPG) of cattleyak. The results showed that the protein encoded by TAF4B did not contain the signal peptide sequence. The expression level of TAF4B in UDSPG of cattleyak was lower than that in yak, while the overexpression of TAF4B in cattleyak promoted the proliferation activity of cattleyak UDSPG. Meanwhile, the expression of proliferation and meiosis-related genes was increased but the differentiation-related genes were decreased. Therefore, the aberrant expression of TAF4B in cattleyak UDSPG possibly impaired its proliferation and differentiation equilibrium and decreased its growth potentiality, thereby reducing the quantity of UDSPG and affecting spermatogenesis. This study provided a potential approach for further elucidation of the mechanism of spermatogenesis arrest and provided a new idea for solving the problem of male infertility in cattleyak.

The success of artificial insemination (AI) with frozen-thawed semen in cattle is influenced by both female factors and sperm quality. In terms of sperm quality, prior studies indicate that the ability of frozen-thawed bovine sperm to fertilise an oocyte is dependent on their quality and resilience to cryopreservation. Cryopreservation induces oxidative stress, leading to ultrastructural damage in the sperm. This study aimed to determine whether the quality of fresh semen can identify bulls with good and poor sperm freezability. This difference between fresh and frozen semen from the same bull allows us to predict fertility. Motility and kinetic parameters were assessed using computer-assisted sperm analysis (CASA), while six functional variables were evaluated through flow cytometry, both before and after the freeze-thaw process on the sperm from 13 bulls. In vivo fertility was measured using 90-day non-return rates. The principal component analysis (PCA) of eight sperm variables post-thaw identified one principal component explaining 81.19% of the total variance and classified the bulls into two groups: Poor freezability bulls (progressive motility: 48.12% ± 8.41%; viability: 77.51% ± 7.61%) and good freezability bulls (progressive motility: 58.64% ± 6.64%; viability: 88.12% ± 2.52%). Bulls with higher freezability showed better sperm viability and motility, as well as lower levels of ROS, superoxides and intracellular calcium before cryopreservation that were significantly correlated with higher non-return rates (NRR). The results underscore the importance of assessing the quality and functionality of fresh semen to predict the fertility potential of cryopreserved sperm. This approach can aid in selecting ejaculates with the best potential for successful artificial insemination, ultimately improving reproductive performance in dairy cattle.

Antioxidants help safeguard sperm cells from damage during the freeze-thaw process. Melatonin and its metabolites have an antioxidative effect. The current study aimed to evaluate the impact of melatonin supplementation in tris-based extenders at various concentrations (0.5, 1.0 and 2 mM) on the freezability of Jamnapari goat spermatozoa. A total of 48 ejaculates were collected twice a week from four Jamnapari bucks (n = 12 × 4) using an artificial vagina during the period of October to November 2023. Selected ejaculates diluted with tris-citric acid egg yolk extender were divided into four equal aliquots, and melatonin (dissolved in 0.1% dimethyl sulfoxide) was added later at 0.5, 1.0 and 2.0 mM to the control group (C) (extender and 0.1% DMSO) and treatment groups T1, T2 and T3, respectively. Various seminal attributes such as progressive motility, livability, acrosomal integrity, sperm abnormalities, sperm plasma membrane integrity, sperm penetration distance by vanguard spermatozoa in polyacrylamide gel and seminal plasma enzyme leakage (AST, ALT, ACP and AKP) were evaluated at post-dilution and post-thawed stages. Our findings revealed that all semen quality parameters were superior in melatonin-treated groups than the C, and the differences were noticeably higher in the T2 group (1.0 mM) than the other groups. Adding 1.0 mM melatonin proved to be the most effective to safeguard sperm cells from cryopreservation induced cryodamage of Jamnapari buck.