Italiya Jignesh, Panchal Ketankumar J, Jakhesara Subhash J, Joshi Chaitanya G, Koringa Prakash G
Department of Veterinary Biotechnology, Veterinary College, Kamdhenu University, Anand, India.
Centre for Infectious Animal Diseases, Faculty of Tropical AgriSciences, Czech University of Life Sciences, Prague, Czechia.
Front Vet Sci. 2024 Jul 12;11:1419573. doi: 10.3389/fvets.2024.1419573. eCollection 2024.
AIM/OBJECTIVES: This study examines the impact of an ethanolic extract derived from seeds on the Gir bull () spermatozoa. The objective is to thoroughly assess the effects of the seed extract on the physiological parameters of bull spermatozoa, followed by evaluating its effects on X and Y-bearing spermatozoa and its impact on gene expression through transcriptome profiling.
For this study, one Gir bull was selected, and 12 ejaculates were collected at one-week time intervals. Sperm cells were isolated from each ejaculate and incubated with varying concentrations of the ethanolic extract. The physiological parameters of the spermatozoa were assessed using Computer Assisted Semen Analysis (CASA) and compared with control groups to evaluate the extract's effects on sperm quality and motility.
At a concentration of 18 mg/mL extract, we noticed a statistically significant 16.4% increase in sperm motility ( = 0.0065). In order to understand the specific effect on X and Y-bearing spermatozoa, motile and non-motile sperm separated by glass wool column method and further evaluated for quantification of X and Y-bearing sperm in all samples by ddPCR. To understand the effect of extract on spermatozoa at the molecular level, whole transcriptome profiling was carried out using Illumina MiSeq. Transcriptome profiling revealed 81 genes that were expressed differently between the group treated with the extract and the control group. The current investigation revealed an increase in the expression of TLX1, CRYGB, KLF13, and ZAR1 transcripts, which play a role in embryonic development. In addition, several genes have been identified that are involved in sperm motility, such GSK3B, LAPRS, MAPK1, CAMK2B, and AQP7. The findings exhibited the therapeutic effectiveness of seeds in augmenting fertility through a synergistic blend of activities, including enhanced sperm motility and positive influence on embryogenesis.
本研究考察从种子中提取的乙醇提取物对吉尔牛精子的影响。目的是全面评估种子提取物对公牛精子生理参数的影响，随后评估其对携带X和Y染色体精子的影响以及通过转录组分析评估其对基因表达的影响。
本研究选取一头吉尔牛，每隔一周采集12次射精样本。从每次射精样本中分离出精子细胞，并与不同浓度的乙醇提取物一起孵育。使用计算机辅助精液分析（CASA）评估精子的生理参数，并与对照组进行比较，以评估提取物对精子质量和活力的影响。
在提取物浓度为18mg/mL时，我们注意到精子活力有统计学意义的16.4%的增加（P=0.0065）。为了解对携带X和Y染色体精子的具体影响，采用玻璃棉柱法分离活动精子和不活动精子，并通过ddPCR进一步评估所有样本中携带X和Y染色体精子的定量情况。为了解提取物在分子水平上对精子的影响，使用Illumina MiSeq进行全转录组分析。转录组分析显示，提取物处理组和对照组之间有81个基因表达存在差异。目前的研究表明，在胚胎发育中起作用的TLX1、CRYGB、KLF13和ZAR1转录本的表达有所增加。此外，还鉴定出了几个与精子活力有关的基因，如GSK3B、LAPRS、MAPK1、CAMK2B和AQP7。研究结果表明，种子通过增强精子活力和对胚胎发生的积极影响等多种活动的协同作用，在提高生育能力方面具有治疗效果。
