Improving litter size by boar spermatozoa: application of combined H33258/CTC staining in field trial with artificial insemination

Kwon, Woo-Sung; Rahman, Saidur; Lee, J.-S.; You, Young Ah; Pang, Myung-Geol

doi:10.1111/andr.12020

Cited by 27 publications

(35 citation statements)

References 33 publications

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“…The capacitation status was determined using the dual staining method described by Kwon et al 12 13 52 . Briefly, 135 μL of treated spermatozoa was added to 15 μL of H33258 solution (10 μg H33258/mL Dulbecco’s PBS [DPBS]) and incubated for 2 min at room temperature (RT).…”

Section: Methodsmentioning

confidence: 99%

Increased male fertility using fertility-related biomarkers

Kwon

Rahman

Ryu

et al. 2015

Sci Rep

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Conventional semen analyses are used to evaluate male factor fertility/infertility in humans and other animals. However, their clinical value remains controversial. Therefore, new tools that more accurately assess male fertility based on sperm function and fertilization mechanism are of interest worldwide. While protein markers in spermatozoa that might help differentiate fertile and infertile sperm have been identified, studies are in their infancy, and the markers require validation in field trials. In the present study, to discover more sensitive biomarkers in spermatozoa for predicting male fertility, we assessed protein expression in capacitated spermatozoa. The results demonstrated that cytochrome b-c1 complex subunit 2 (UQCRC2) was abundantly expressed in high-litter size spermatozoa (>3-fold). On the other hand, equatorin, beta-tubulin, cytochrome b-c1 complex subunit 1 (UQCRC1), speriolin, Ras-related protein Rab-2A (RAB2A), spermadhesin AQN-3, and seminal plasma sperm motility inhibitor were abundantly expressed in low-litter size spermatozoa (>3-fold). Moreover, RAB2A and UQCRC1 expression negatively correlated with litter size, while UQCRC2 expression positively correlated with litter size. Finally, the putative biomarkers predicted litter size in field trials. Our study suggests that biomarkers present in spermatozoa after capacitation can help differentiate superior male fertility from below-average fertility with high sensitivity.

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Section: Methodsmentioning

confidence: 99%

Increased male fertility using fertility-related biomarkers

Kwon

Rahman

Ryu

et al. 2015

Sci Rep

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“…Yorkshire and Duroc semen samples were collected using the gloved‐hand technique twice per week (Almond et al ., ). The lower limit values for AI were calculated following criteria previously described (Kwon et al ., 2015c). Semen samples were diluted with commercial extender (Beltsville Thawing Solution) for AI (3 × 10 9 sperm cells/100 mL).…”

Section: Methodsmentioning

confidence: 99%

“…Three different ejaculated semen samples ( n = 3) from 20 randomly selected Yorkshire (average litter size 11.77 ± 0.24) and Duroc (average litter size 8.54 ± 0.42) boars were washed to remove seminal plasma and dead spermatozoa with a discontinuous [70% (v/v) and 35% (v/v)] Percoll gradient (Sigma, St Louis, MO, USA) at 500 g for 20 min (Flesch et al ., ; Kwon et al ., ,b). Capacitation was induced as previously described (Kwon et al ., 2014b, ,d). To induce capacitation, samples were incubated with modified tissue culture medium 199 [containing 10% fetal bovine serum (v/v), 0.91 m m sodium pyruvate, 3.05 m m d ‐glucose, 2.92 m m calcium lactate, 2.2 g/L sodium bicarbonate, and 10 μg/mL heparin; Sigma) for 30 min at 37 °C under an atmosphere of 5% CO 2 in air.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, the pooled semen samples from Yorkshire and Duroc boar were washed with a discontinuous [70% (v/v) and 35% (v/v)] Percoll gradient at 500 g for 20 min. Capacitation was induced as previously described (Kwon et al ., 2014b, ,d). Capacitation‐induced samples were washed once more with a discontinuous Percoll gradient to remove media for inducing of capacitation.…”

Section: Methodsmentioning

confidence: 99%

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Comparison of markers predicting litter size in different pig breeds

et al. 2017

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To overcome the limitations of conventional analysis of male fertility in animals and humans, proteomic studies have been performed to develop fertility-related biomarkers for prognosis and diagnosis of male fertility. However, the studies were focused on specific species or breeds. Therefore, a study is required to validate whether fertility-related markers would apply to other breeds in pigs. In this study, previously developed fertility-related biomarkers from Landrace were validated to use for prognosis of male fertility in commercially available breeds. Expression level of eight biomarkers in non-capacitated and capacitated (C) spermatozoa from Yorkshire and Duroc boars was analyzed. And then, to explore the validity of these markers for prognosis of male fertility, i.e. litter size, artificial insemination was performed. Among them, RAB2A (NC) and UQCRC1 (NC) turned out to be highest efficient markers for Yorkshire. RAB2A (C) was most efficient marker for Duroc. Average litter size has increased as much as 1.41 live born after prediction using eight fertility-related biomarkers in Yorkshire. In addition, average 2.52 litter size was increased after prediction using eight fertility-related biomarkers in Duroc. Average litter sizes were especially highly increased after prediction of fertility using RAB2A (NC) in Yorkshire (1.57 piglets) and TPI (NC) in Duroc (3.14 piglets), respectively. As a result, all biomarkers were significantly correlated with litter size. However, overall accuracy to predict litter size in three breeds was different in response with each marker. Average litter size after artificial insemination was also significantly affected by marker selection. Therefore, this study suggests that developed fertility-related markers may be used for prognosis and diagnosis of male fertility irrespective of breed. However, selection of efficient markers for breeds should be considered to obtain more accurate and efficient outcomes.

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“…Combined Hoechst 33258/chlortetracycline fluorescence (H33258/CTC) staining is widely used method to directly measure the capacitation status of spermatozoa by monitoring calcium-regulated changes using the fluorescent antibiotic chlortetracycline [ 12 , 13 ]. Recently, it has been reported that the H33258/CTC staining has been applied for evaluating boar fertility in order to overcome the limitations of current semen analyses [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Applications of capacitation status for litter size enhancement in various pig breeds

Kwon

Shin

Ryu

et al. 2018

Asian-Australas J Anim Sci

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ObjectiveSeveral studies have reported the development of new molecular methods for the prognosis and diagnosis of male fertility based on biomarkers aimed at overcoming the limitations of conventional male fertility analysis tools. However, further studies are needed for the field application of these methods. Therefore, alternative methods based on existing semen analysis methods are required to improve production efficiency in the animal industry.Methodswe examined the possibility of improving litter size in various pig breeds using combined Hoechst 33258/chlortetracycline fluorescence (H33258/CTC) staining. The correlation between field fertility and capacitation status by combined H33258/CTC staining in different ejaculates spermatozoa (n = 3) from an individual boar (20 Landrace, 20 Yorkshire, and 20 Duroc) was evaluated as well as overall accuracy.ResultsThe acrosome reacted (AR) pattern after capacitation (%) was positively correlated with the litter size of Landrace, Yorkshire, and Duroc pigs and the overall accuracy was 75%, 75%, and 70% in Landrace, Yorkshire, and Duroc pigs, respectively. The difference (Δ) in AR pattern before and after capacitation was positively correlated with the litter size of Landrace, Yorkshire, and Duroc pigs and the overall accuracy was 80%, 65%, and 55% in Landrace, Yorkshire, and Duroc pigs, respectively. However, the difference (Δ) in capacitated (B) pattern before and after capacitation was negatively correlated with the litter size of Landrace pigs and the overall accuracy was 75%. Moreover, average litter size was significantly altered according to different combined H33258/CTC staining parameters.ConclusionThese results show that combined H33258/CTC staining may be used to predict male fertility in various breeds. However, the selection of specific efficiency combined H33258/CTC staining parameters requires further consideration. Taken together, these findings suggest that combined H33258/CTC staining may constitute an alternative method for predicting male fertility until such time as fertility-related biomarkers are further validated.

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Improving litter size by boar spermatozoa: application of combined H33258/CTC staining in field trial with artificial insemination

Cited by 27 publications

References 33 publications

Increased male fertility using fertility-related biomarkers

Increased male fertility using fertility-related biomarkers

Comparison of markers predicting litter size in different pig breeds

Applications of capacitation status for litter size enhancement in various pig breeds

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