TLR7/8 agonist (R848) inhibit bovine X sperm motility via PI3K/GSK3α/β and PI3K/NFκB pathways

Wen, Fei; Liu, Weidong; Liu, Yu; Zou, Qian-Long; Xian, Ming; Han, Shuaiqi; Zhang, Hongyun; Liu, Songqi; Feng, Xianzou; Hu, Jianhong

doi:10.1016/j.ijbiomac.2023.123485

Cited by 4 publications

(1 citation statement)

References 33 publications

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“…In 2022, Huang [ 20 ] combined R848 with an alkaline semen diluent, achieving the successful isolation and enrichment of goat X sperm. This sex control technique, relying on the R848 activation of X sperm TLR7/8, was successfully validated in several mammals, including mice, cattle, and sheep, confirming the robustness of this sex control technology [ 17 , 18 , 19 , 20 , 21 ].…”

Section: Introductionmentioning

confidence: 80%

Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm via Ligands dsRNA-40 and dsRNA-DR

Hou,

Peng,

Hong

et al. 2024

Molecules

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Gender control technologies are promising for enhancing the production efficiency of the farm animal industry, and preventing sex-linked hereditary diseases in humans. It has been shown that the X sperm of mammalian animals specifically expresses X-chromosome-derived toll-like receptor 7/8 (TLR7/8), and the activation of TLR7/8 on the X sperm by their agonist, R848, can separate X and Y sperm via the specific inhibition of X sperm motility. The use of R848-preselected sperm for fertilization resulted in sex-ratio-skewed embryos or offspring. In this study, we aimed to investigate whether two other TLR7/8 ligands, double-stranded RNA-40 (dsRNA-40) and double-stranded RNA-DR (dsRNA-DR), are also effective in the separation of mouse X and Y sperm and the subsequent generation of gender-ratio-skewed in vitro fertilization (IVF) embryos. Our results indicated that cholesterol modification significantly enhances the transfection of dsRNA-40 and dsRNA-DR into sperm cells. dsRNA-40 and dsRNA-DR incubation with mouse sperm could separate X and Y sperm by the specific suppression of X sperm motility by decreasing its ATP level and mitochondrial activity. The use of a dsRNA-40- or dsRNA-DR-preselected upper layer of sperm, which predominantly contains high-motility Y sperm, for IVF caused a male-biased sex ratio shift in resulting embryos (with 65.90–74.93% of embryos being male). This study develops a simple new method for the efficient separation of mammalian X and Y sperm, enabling the selective production of male or female progenies.

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Section: Introductionmentioning

confidence: 80%

Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm via Ligands dsRNA-40 and dsRNA-DR

Hou,

Peng,

Hong

et al. 2024

Molecules

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Proteomic and Metabolomic Profiling Reveals Alterations in Boar X and Y Sperm

Cheng,

Hao,

Zhang

et al. 2024

Animals

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Sex-controlled sperm combined with artificial insemination allows animals to reproduce offspring according to the desired sex, accelerates the process of animal genetics and breeding and promotes the development of animal husbandry. However, the molecular markers for sexual sperm sorting are unusual. To identify the molecular markers of boar sperm sorting, proteomics and metabolomics techniques were applied to analyze the differences in proteins and metabolism between X and Y sperm. Label-free quantitative proteomics identified 254 differentially expressed proteins (DEPs) in the X and Y sperm of boars, including 106 proteins that were highly expressed in X sperm and 148 proteins that were highly expressed in Y sperm. Among the differential proteins, COX6A1, COX1, CYTB, FUT8, GSTK1 and PFK1 were selected as potential biological markers for X and Y sperm sorting. Moreover, 760 metabolites from X and Y sperm were detected. There were 439 positive ion mode metabolites and 321 negative ion mode metabolites identified. The various metabolites were phosphoenolpyruvate, phytosphingosine, L-arginine, N-acetylputrescine, cytidine-5′-diphosphate and deoxyuridine. These metabolites were mainly involved in the TCA cycle, oxidative phosphorylation pathway, glycolysis pathway, lipid metabolism pathway, amino acid metabolism pathway, pentose phosphate pathway and nucleic acid metabolism pathway. The differential proteins and differential metabolites obtained by the combined proteomics and metabolomics analysis were projected simultaneously to the KEGG pathway, and a total of five pathways were enriched, namely oxidative phosphorylation pathway, purine metabolism, unsaturated fatty acid biosynthesis, ABC transporters and peroxisomes. In summary, COX6A1 and CYTB were identified as potential biomarkers for boar X and Y sperm sorting.

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Reproductive Tract Mucus May Influence the Sex of Offspring in Cattle: Study in Cows That Have Repeatedly Calved Single-Sex Offspring

Huang,

Niu,

Wang

et al. 2024

Veterinary Sciences

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This study aimed to investigate the selective effect of the reproductive tract mucus in cows that have consistently produced offspring of a single sex on X/Y spermatozoa. We collected mucus from the reproductive tract of cows that had given calvings to offspring of the same sex, or alternated between sexes, for more than five consecutive calvings. We evaluated the pH of reproductive tract mucus. Subsequently, we conducted a spermatozoa penetration assay; the proportions of X and Y spermatozoa after penetration were then identified by dual TaqMan qPCR and flow cytometry. This was followed by in vitro fertilization and embryo sex determination experiments. Immediately afterwards, computer-aided spermatozoa analysis was employed to analyze the spermatozoa that had penetrated through different types of mucus in the reproductive tract. The analysis indicated that the reproductive tract mucus of cows consistently producing male or female calves exhibited selectivity towards X/Y spermatozoa. The differences in the pH values of the reproductive tract mucus among cows continuously producing male calves, those continuously producing female calves, and those alternately giving birth to male and female calves were not significant (p ≥ 0.05). The outcome of dual TaqMan qPCR for cows consistently producing male calves was Y: 79.29 ± 4.28% vs. X: 21.67 ± 4.53%; for cows consistently producing female calves, the equation was Y: 25.05 ± 4.88% vs. X: 75.34 ± 5.13%. The results of flow cytometry processing revealed the following proportions: for cows consistently producing male calves: Y: 83.33 ± 5.52% vs. X: 17.23 ± 4.74%; for cows consistently producing female calves: Y: 24.81 ± 4.13% vs. X: 76.64 ± 4.21%. The outcomes of embryo sex determination for cows consistently producing male calves were as follows: male embryos vs. female embryos (79.60 ± 2.87% vs. 21.07 ± 2.51%); for cows consistently producing female calves, the outcomes for male embryos vs. female embryos were 25.58 ± 3.96% vs. 75.63 ± 3.55%. Computer-aided analysis revealed that the concentration of spermatozoa penetrating the reproductive tract mucus in cows alternating between male and female calves (9.09 ± 0.72 million/mL) was significantly higher than that in cows consistently producing male calves (6.01 ± 1.19 million/mL) and cows consistently producing female calves (5.61 ± 0.60 million/mL). There were no significant differences in spermatozoa motility, the proportion of progressive motile spermatozoa, and curvilinear, straight-line, and average path velocities. Collectively, these findings indicate that the reproductive tract mucus of cows consistently producing offspring of a single sex exhibits selectivity towards either X or Y spermatozoa. This finding is of great significance for studying the impact of maternal factors on offspring sex.

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TLR7/8 agonist (R848) inhibit bovine X sperm motility via PI3K/GSK3α/β and PI3K/NFκB pathways

Cited by 4 publications

References 33 publications

Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm via Ligands dsRNA-40 and dsRNA-DR

Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm via Ligands dsRNA-40 and dsRNA-DR

Proteomic and Metabolomic Profiling Reveals Alterations in Boar X and Y Sperm

Reproductive Tract Mucus May Influence the Sex of Offspring in Cattle: Study in Cows That Have Repeatedly Calved Single-Sex Offspring

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