AIM/OBJECTIVES: This study examines the in vitro impact of an ethanolic extract derived from Bryonia laciniosa seeds on the Gir bull (Bos indicus) 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. MATERIAL METHOD: 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. RESULTS AND DISCUSSION: At a concentration of 18 mg/mL B. laciniosa extract, we noticed a statistically significant 16.4% increase in sperm motility (p = 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 B. laciniosa 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 B. laciniosa seeds in augmenting fertility through a synergistic blend of activities, including enhanced sperm motility and positive influence on embryogenesis.

Centre for Infectious Animal Diseases Faculty of Tropical AgriSciences Czech University of Life Sciences Prague Czechia

Department of Science and Technology Gujarat Biotechnology Research Centre Government of Gujarat Gandhinagar India

Department of Veterinary Biotechnology Veterinary College Kamdhenu University Anand India

P D Patel Institute of Applied Sciences Charotar University of Science and Technology Changa India

Zobrazit více v PubMed

Peddinti D, Nanduri B, Kaya A, Feugang JM, Burgess SC, Memili E. Comprehensive proteomic analysis of bovine spermatozoa of varying fertility rates and identification of biomarkers associated with fertility. BMC Syst Biol. (2008) 2:1–13. doi: 10.1186/1752-0509-2-19, PMID: PubMed DOI PMC

Butler ML, Bormann JM, Weaber RL, Grieger DM, Rolf MM. Selection for bull fertility: a review. Transl Anim Sci. (2020) 4:423–41. doi: 10.1093/tas/txz174, PMID: PubMed DOI PMC

Kadam PV, Bhapkar PV, Shaikh SK, Yadav KN, Giram DK, Karanje AS. Bryonia laciniosa: a ethnopharmacological approach of ayurvedic Shivlingi. Pharmacogn Res. (2023) 15:462–7. doi: 10.5530/pres.15.3.047 DOI

Lessard C, Siqueira L, D’amours O, Sullivan R, Leclerc P, Palmer C. Infertility in a beef bull due to a failure in the capacitation process. Theriogenology. (2011) 76:891–9. doi: 10.1016/j.theriogenology.2011.04.020 PubMed DOI

Parrish J, Susko-Parrish J, Winer M, First N. Capacitation of bovine sperm by heparin. Biol Reprod. (1988) 38:1171–80. doi: 10.1095/biolreprod38.5.1171 PubMed DOI

Wang M-Y, West BJ, Jensen CJ, Nowicki D, Su C, Palu AK, et al. . Morinda citrifolia (Noni): a literature review and recent advances in Noni research. Acta Pharmacol Sin. (2002) 23:1127–41. PubMed

Baliga MS. Triphala, ayurvedic formulation for treating and preventing cancer: a review. J Altern Complement Med. (2010) 16:1301–8. doi: 10.1089/acm.2009.0633 PubMed DOI

Gabrielian S, Gevorgovich A. Bryonia, as novel plant adoptogen, for prevention and treatment of stress induced disorders. Promising Res Abstr. (1997) 5003:1–8.

Kaurav H, Choudhary S, Chaudhary G. Bryonia laciniosa an ayurvedic herbal plant ‘Bryonia laciniosa’ with its ethnomedicinal significance. J Drug Deliv Therap. (2021) 11:137–41. doi: 10.22270/jddt.v11i3-S.4889 DOI

Sud K, Sud S. A scientific review on Shivlingi Beej (Bryonia Laciniosa): a mystical ethno-medicine for infertility. Eur J Biomed Pharm Sci. (2017) 4:1098–102.

Ros-Santaella JL, Pintus E. Plant extracts as alternative additives for sperm preservation. Antioxidants. (2021) 10:772. doi: 10.3390/antiox10050772 PubMed DOI PMC

Truong VB, Davis OS, Gracey J, Neal MS, Khokhar JY, Favetta LA. Sperm capacitation and transcripts levels are altered by in vitro THC exposure. BMC Mol Cell Biol. (2023) 24:1–13. doi: 10.1186/s12860-023-00468-3 PubMed DOI PMC

Parrish JJ. Bovine in vitro fertilization: in vitro oocyte maturation and sperm capacitation with heparin. Theriogenology. (2014) 81:67–73. doi: 10.1016/j.theriogenology.2013.08.005, PMID: PubMed DOI PMC

Van Der Ven H, Jeyendran R, Al-Hasani S, Tünnerhoff A, Hoebbel K, Diedrich K, et al. . Glass wool column filtration of human semen: relation to swim-up procedure and outcome of IVF. Hum Reprod. (1988) 3:85–8. doi: 10.1093/oxfordjournals.humrep.a136659, PMID: PubMed DOI

Pathak P, Dhami A, Chaudhari D, Hadiya K. Motion characteristics and kinematics of fresh spermatozoa of Gir, Surti and Murrah bulls assessed by computer assisted semen analyzer. Indian J Vet Sci Biotechnol. (2019) 14:14–9. doi: 10.21887/ijvsbt.14.3.4 DOI

Panda H. Handbook on herbal medicines. Delhi, India: Asia Pacific Business Press Inc. (2004).

Vaidya L. Bhavprakash. Varanasi: Motilal Banarsidass; (1970).

Chauhan NS, Dixit VK. Effects of Bryonia laciniosa seeds on sexual behaviour of male rats. Int J Impot Res. (2010) 22:190–5. doi: 10.1038/ijir.2009.62 PubMed DOI

Italiya JM, Patel MR, Golaviya AV, Patel SS, Thakkar BK, Jakhesara SJ, et al. . RNA-sequencing attest increased sperm motility in bovine spermatozoa treated with ethanolic extract of Putranjiva roxburghii. 3 Biotech. (2023) 13:33. doi: 10.1007/s13205-022-03452-4 PubMed DOI PMC

Gur Y, Breitbart H. Mammalian sperm translate nuclear-encoded proteins by mitochondrial-type ribosomes. Genes Dev. (2006) 20:411–6. doi: 10.1101/gad.367606 PubMed DOI PMC

Rajamanickam GD, Kastelic JP, Thundathil JC. Content of testis-specific isoform of Na/K-Atpase (Atp1A4) is increased during bovine sperm capacitation through translation in mitochondrial ribosomes. Cell Tissue Res. (2017) 368:187–200. doi: 10.1007/s00441-016-2514-7, PMID: PubMed DOI

Wu X, Viveiros MM, Eppig JJ, Bai Y, Fitzpatrick SL, Matzuk MM. Zygote arrest 1 (Zar1) is a novel maternal-effect gene critical for the oocyte-to-embryo transition. Nat Genet. (2003) 33:187–91. doi: 10.1038/ng1079, PMID: PubMed DOI

Chen L, Yao F, Qin Y, Shao Y, Fang L, Yu X, et al. . The potential role of Krüppel-like factor 13 (Aj-klf13) in the intestine regeneration of sea cucumber Apostichopus japonicus. Gene. (2020) 735:144407. doi: 10.1016/j.gene.2020.144407, PMID: PubMed DOI

Zhu Z, Li R, Wang L, Zheng Y, Hoque SM, Lv Y, et al. . Glycogen synthase kinase-3 regulates sperm motility and acrosome reaction via affecting energy metabolism in goats. Front Physiol. (2019) 10:968. doi: 10.3389/fphys.2019.00968 PubMed DOI PMC

Liao H-Y, O’flaherty C. Lysophosphatidic acid signalling regulates human sperm viability via the phosphoinositide 3-kinase/Akt pathway. Cells. (2023) 12:2196. doi: 10.3390/cells12172196 PubMed DOI PMC

Li Y, Jin L, Li Y, Qian J, Wang Z, Zheng X, et al. . Lysophosphatidic acid improves human sperm motility by enhancing glycolysis and activating L-type calcium channels. Front Endocrinol. (2022) 13:896558. doi: 10.3389/fendo.2022.896558, PMID: PubMed DOI PMC

Chen Y, Wang K, Zhang D, Zhao Z, Huang J, Zhou L, et al. . Gpx6 is involved in the in vitro induced capacitation and acrosome reaction in porcine sperm. Theriogenology. (2020) 156:107–15. doi: 10.1016/j.theriogenology.2020.06.020 PubMed DOI

Schlingmann K, Michaut MA, Mcelwee JL, Wolff CA, Travis AJ, Turner RM. Calmodulin and Camkii in the sperm principal piece: evidence for a motility-related calcium/calmodulin pathway. J Androl. (2007) 28:706–16. doi: 10.2164/jandrol.106.001669, PMID: PubMed DOI

Saito K, Kageyama Y, Okada Y, Kawakami S, Kihara K, Ishibashi K, et al. . Localization of aquaporin-7 in human testis and ejaculated sperm: possible involvement in maintenance of sperm quality. J Urol. (2004) 172:2073–6. doi: 10.1097/01.ju.0000141499.08650.ab, PMID: PubMed DOI