Freezing-thawing induces deprotamination, cryocapacitation-associated changes; DNA fragmentation; and reduced progesterone sensitivity in buck spermatozoa

Kritaniya, Deepika; Yadav, Sarita; Swain, Dilip Kumar; Reddy, A. Vidyasagar; Dhariya, Rahul; Yadav, Brijesh; Anand, Mukul; Nigam, Rajesh

doi:10.1016/j.anireprosci.2020.106628

Cited by 9 publications

(25 citation statements)

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“…The live spermatozoa having intact acrosome were evaluated using the fluorescent FITC‐PSA‐PI dual staining using the standard procedures previously described (Mendoza et al., 1992) and recently modified and reported from our group (Kritaniya et al., 2020). Slides were examined under an EVOS FL inverted digital fluorescent microscope (Invitrogen, USA) FITC/blue (Green Fluorescent Filter) and PI/green filters (Red Fluorescent Filter), and the images were overlaid to count different cell types.…”

Section: Methodsmentioning

confidence: 99%

“…The localisation of progesterone receptor was conducted using P-BSA-FITC assay as previously described by Cheng et al (1998) and recently reported in goat spermatozoa by Kritaniya et al (2020). In the assay, P-BSA acts as a conjugate ligand for binding to progesterone membrane receptors.…”

Section: Progesterone Receptor Binding (Progesterone 3-(o-carboxymeth...mentioning

confidence: 99%

“…After 24 hr of freezing in liquid N2, semen straws were thawed using a thawing unit (Minitube, Tifenberg, Germany) at 37℃ for 45 s and were evaluated for various sperm parameters using standard techniques as established in our laboratory (Kritaniya et al, 2020;Shah et al, 2017;Swain et al, 2017;Yadav et al, 2017Yadav et al, , 2019.…”

Section: Post-thaw Evaluation Of the Semenmentioning

confidence: 99%

“…Further, immunolocalisation of tyrosine-phosphorylated proteins was carried out using indirect immunofluorescence as described previously (Kritaniya et al, 2020;Shah et al, 2017;Yadav et al, 2017).…”

Section: Immunoblotting and Immunolocalisation Of Tyrosine-phosphoryl...mentioning

confidence: 99%

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Curcumin in a tris‐based semen extender improves cryosurvival ofHarianabull spermatozoa

et al. 2021

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In this study, the cryoprotective potential of natural antioxidant curcumin in Hariana bull semen was evaluated as an additive in a tris‐based extender with the assessment of motility and motion parameters of spermatozoa, membrane intactness, progesterone‐receptor binding, protein carbonyl content, cervical mucus penetration, cryocapacitation‐associated and apoptotic‐like changes. The collected ejaculates were divided into five groups in the tris‐based extender (control without curcumin‐I, 10 µM‐II, 25 µM‐III, 50 µM‐IV and 75µM‐V) and were cryopreserved. Groups II and III containing 10 and 25 µM curcumin substantially (p < .05) improved the post‐thaw sperm parameters like viability, motility, and velocity parameters; intact acrosome and membrane; lowered protein carbonyl content; DNA fragmentation and cryocapacitation‐associated changes in comparison to control. It was interesting to note that early apoptotic‐like changes in sperm cells were significantly (p < .05) decreased in Group II along with an increase in a higher population of sperm cells having high mitochondrial transmembrane potential. Higher progesterone‐receptor binding, Vanguard distance and in vitro capacitation response were observed only in Group II (10µM) compared to other groups. In conclusion, curcumin in a semen extender manifests cryoprotective effects and may be incorporated at 10 µM concentration in a Hariana bull semen extender for better post‐thaw sperm quality.

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Section: Methodsmentioning

confidence: 99%

Section: Progesterone Receptor Binding (Progesterone 3-(o-carboxymeth...mentioning

confidence: 99%

Section: Post-thaw Evaluation Of the Semenmentioning

confidence: 99%

Section: Immunoblotting and Immunolocalisation Of Tyrosine-phosphoryl...mentioning

confidence: 99%

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Curcumin in a tris‐based semen extender improves cryosurvival ofHarianabull spermatozoa

et al. 2021

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“…However, little information is present in the literature regarding the effects of cryopreservation or capacitation on sperm chromatin condensation. In this sense, a recent study reported that chromatin deprotamination occurs during sperm cryopreservation in bucks [ 37 ], probably due to capacitation-like changes. Moreover, another study reported that the sperm chromatin of bulls showing high NRR was able to better withstand the decondensation induced by EDTA and SDS [ 38 ].…”

Section: Discussionmentioning

confidence: 99%

Sperm chromatin condensation as an in vivo fertility biomarker in bulls: a flow cytometry approach

Llavanera

Ribas-Maynou

Delgado-Bermúdez

et al. 2021

J Animal Sci Biotechnol

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Background Genetic selection in cattle has been directed to increase milk production. This, coupled to the fact that the vast majority of bovine artificial inseminations (AI) are performed using cryopreserved sperm, have led to a reduction of fertility rates over the years. Thus, seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency. In humans, sperm chromatin condensation evaluated through chromomycin A3 (CMA3) has recently been purported to be a powerful biomarker for sperm functional status and male infertility. The objectives of the present study were: a) to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability, and b) to test whether this parameter could be used as a predictor of in vivo fertility in bulls. The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males. Reproductive outcomes of each sire were determined by non-return rates, which were used to classify bulls into two groups (highly fertile and subfertile). Results Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry. Sperm quality parameters (morphology, viability, total and progressive motility) were also assessed. Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility. Sperm morphology, viability and total motility presented an area under the ROC curve (AUC) of 0.54, 0.64 and 0.68, respectively (P > 0.05), and thus were not able to discriminate between fertile and subfertile individuals. Alternatively, while the percentage of progressively motile sperm showed a significant predictive value, with an AUC of 0.73 (P = 0.05), CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls. Specifically, the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility, with an AUC of 0.78 (P = 0.02). Conclusions Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility, with a potential ability to maximising the efficiency of dairy breeding industry.

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Interplay mechanisms between progesterone and endocannabinoid receptors in regulating bull sperm capacitation and acrosome reaction

Mahajan

Sharma

Mishra

et al. 2022

Journal Cellular Physiology

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After ejaculation, sperm show a limited capacity for transcription and translation. In the oviduct, most of the signalling in sperm is nongenomic and is mediated through membrane receptors. Studies have shown that the cation channel of sperm (CatSper), cAMP, cGMP, protein kinases, and tyrosine phosphorylation are involved in the nongenomic signalling of progesterone (P4) in sperm. However, it is not known whether there is an interplay between P4 and cannabinoid receptors 1 and 2 (CB1 and CB2), transient receptor potential vanilloid 1 (TRPV1), CatSper channels, cAMP, inositol trisphosphate receptor (IP3R), and mitogen-activated protein kinase (MAPK); these potential regulators are involved in the regulation of capacitation and the acrosome reaction. In the present study, selective blockers of CB1, CB2, TRPV1, CatSper channels, cAMP, protein kinase A (PKA), IP3R, and MAPK were used to identify their involvement in P4-mediated bull sperm capacitation and the acrosome reaction. Selective blocking of any one of the molecules caused a significant reduction in P4 signalling (p < 0.05). Interestingly, blocking these molecules in combination followed by treatment with P4 resulted in the complete absence of capacitation and the acrosome reaction. Blocking a single receptor was not able to eliminate the P4-induced capacitation and the acrosome reaction. In addition to the CB1 and CB2 receptors, there may be other signalling pathways that mediate P4 signalling. In conclusion, P4 signalling exhibited interplay with the cannabinoid receptors. The regulation of sperm capacitation and the acrosome reaction also involved cAMP, PKA, L-type and T-type calcium channels, TRPV1, inositol trisphosphate, and MAPK.

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Freezing-thawing induces deprotamination, cryocapacitation-associated changes; DNA fragmentation; and reduced progesterone sensitivity in buck spermatozoa

Cited by 9 publications

References 51 publications

Curcumin in a tris‐based semen extender improves cryosurvival ofHarianabull spermatozoa

Curcumin in a tris‐based semen extender improves cryosurvival ofHarianabull spermatozoa

Sperm chromatin condensation as an in vivo fertility biomarker in bulls: a flow cytometry approach

Interplay mechanisms between progesterone and endocannabinoid receptors in regulating bull sperm capacitation and acrosome reaction

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