Commercial application of flow cytometry for evaluating bull sperm

DeJarnette, J.M.; Harstine, B.R.; McDonald, Kristina; Marshall, C.E.

doi:10.1016/j.anireprosci.2021.106838

Cited by 13 publications

(13 citation statements)

References 20 publications

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“…A washing step to exclude extender components before staining may minimize artefacts but as the impact of the extender on the analyses was of major interest, no washing steps were performed. As shown in this study, DeJarnette et al (2021) also pointed out the importance of establishing adapted laboratory protocols for different extenders in order to obtain correct and comparable results.…”

Section: Discussionmentioning

confidence: 53%

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Assessment of the stallion sperm acrosome in two different extenders: Spermac stain in comparison with PNA/PSA/PI triple‐staining

Dierberger,

Pieper,

Jung

et al. 2023

Reprod Domestic Animals

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Since the stallion acrosome is very small compared to other species and cannot be properly assessed without additional staining, several labelling techniques were developed to facilitate its assessment. The aim of this study was to compare the Spermac stain (Minitüb GmbH) and a PNA/PSA/PI triple‐staining detected by flow cytometry with regard to method agreement for detecting non‐intact acrosomes within two different extenders. For this purpose, eighteen stallion ejaculates were split in half and diluted with the semen extenders EquiPlus or Gent (Minitüb GmbH) to a final concentration of 50 × 106 sperm/mL, respectively. Subsequently, 126 semen samples were stained with both methods between 4 and 240 h (mean: 63.8 ± 48.9 h) after semen collection. Calculated Intraclass correlation coefficients revealed excellent correlations between both methods for EquiPlus (r = .77, p < .001) and fair correlations for Gent (r = .49, p < .001). Interestingly, flow cytometry detected more non‐intact acrosomes in EquiPlus than in Gent (p < .001), whereas the Spermac stain showed no differences (p = .902) between extenders. The poorer method agreement in Gent could be caused by egg yolk artefacts, which made interpretation difficult, so flow cytometry might be preferred. The differences in detected non‐intact acrosomes between extenders highlighted the importance of establishing adapted laboratory protocols for different extender types in order to generate comparable results.

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Section: Discussionmentioning

confidence: 53%

“…As shown in this study, DeJarnette et al (2021) also pointed out the importance of establishing adapted laboratory protocols for different extenders in order to obtain correct and comparable results.…”

Section: Discussionmentioning

confidence: 57%

Assessment of the stallion sperm acrosome in two different extenders: Spermac stain in comparison with PNA/PSA/PI triple‐staining

Dierberger,

Pieper,

Jung

et al. 2023

Reprod Domestic Animals

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“…Despite improvements in recent years in the in vitro analysis of semen, both in the standardization and inclusion of multiple variables in the mathematical models, the predictive capacity of these in vitro assessments continues to be limited. 42 On the one hand, both CASA and FC procedures allow us to analyze a large number of spermatozoa in a more objective way in order to explain part of fertility variation. 7 On the other hand, proteomic analyses allow us to identify potential biomarkers associated with sperm quality and fertility.…”

Section: Discussionmentioning

confidence: 99%

“…Predicting the fertility potential of males or seminal samples is a challenge that has not yet been resolved. Despite improvements in recent years in the in vitro analysis of semen, both in the standardization and inclusion of multiple variables in the mathematical models, the predictive capacity of these in vitro assessments continues to be limited . On the one hand, both CASA and FC procedures allow us to analyze a large number of spermatozoa in a more objective way in order to explain part of fertility variation .…”

Section: Discussionmentioning

confidence: 99%

Bull Sperm SWATH-MS-Based Proteomics Reveals Link between High Fertility and Energy Production, Motility Structures, and Sperm–Oocyte Interaction

Gacem,

Castello-Ruiz,

Hidalgo

et al. 2023

J. Proteome Res.

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The prediction of male or semen fertility potential remains a persistent challenge that has yet to be fully resolved. This work analyzed several in vitro parameters and proteome of spermatozoa in bulls cataloged as high-(HF; n = 5) and low-field (LF; n = 5) fertility after more than a thousand artificial inseminations. Sperm motility was evaluated by computer-assisted sperm analysis. Sperm viability, mitochondrial membrane potential (MMP) and reactive oxygen species (mROS) of spermatozoa were assessed by flow cytometry. Proteome was evaluated by the SWATH-MS procedure. Spermatozoa of HF bulls showed significantly higher total motility than the LF group (41.4% vs 29.7%). Rates of healthy sperm (live, high MMP, and low mROS) for HF and LF bull groups were 49% and 43%, respectively (p > 0.05). Spermatozoa of HF bulls showed a higher presence of differentially abundant proteins (DAPs) related to both energy production (COX7C), mainly the OXPHOS pathway, and the development of structures linked with the motility process (TPPP2, SSMEM1, and SPAG16). Furthermore, we observed that equatorin (EQTN), together with other DAPs related to the interaction with the oocyte, was overrepresented in HF bull spermatozoa. The biological processes related to protein processing, catabolism, and protein folding were found to be overrepresented in LF bull sperm in which the HSP90AA1 chaperone was identified as the most DAP. Data are available via ProteomeXchange with identifier PXD042286.

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“…Subfertility, a signi cantly reduced reproduction rate relative to average reproduction rate by most other bulls, is a major obstacle to the pro tability of the U.S. meat and dairy industry (Shalloo et al, 2014). Currently, the markers used to diagnose bull subfertility explain ~ 3% of the total variance in arti cial insemination eld fertility deviations (DeJarnette et al, 2022). Bovine sperm is extensively analyzed prior to use and undiagnosed subfertility after analysis points to an unknown male factor and a gap in our understanding of sperm biology.…”

Section: Introductionmentioning

confidence: 99%

Abnormal Centriolar Biomarker Ratios Correlate with Unexplained Bull Artificial Insemination Subfertility – a Pilot Study

Turner

Achinger

Kong

et al. 2023

Preprint

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The mechanisms underlying male infertility are poorly understood. Most mammalian spermatozoa have two centrioles: the typical barrel-shaped proximal centriole (PC) and the atypical fan-like distal centriole (DC) connected to the axoneme (Ax). These structures are essential for fertility. However, the relationship between centriole quality and subfertility (reduced fertility) is not well established. Here, we tested the hypothesis that assessing sperm centriole quality can identify cattle subfertility. By comparing sperm from 25 fertile and 6 subfertile bulls, all with normal semen analyses, we found that unexplained subfertility and lower sire conception rates (pregnancy rate from artificial insemination in cattle) corelate with abnormal centriolar biomarker distribution. Fluorescence-based Ratiometric Analysis of Sperm Centrioles (FRAC) found only four fertile bulls (4/25, 16%) had positive FRAC tests (having one or more mean FRAC ratios outside of the distribution range in a group’s high-quality sperm population), whereas all of the subfertile bulls (6/6, 100%) had positive FRAC tests (P=0.00008). The most sensitive biomarker was Acetylated Tubulin, which had a novel labeling pattern between the DC and Ax. These data suggest that FRAC and Acetylated Tubulin labeling can identify bull subfertility that remains undetected by current methods and may provide insight into a novel mechanism of subfertility.

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Commercial application of flow cytometry for evaluating bull sperm

Cited by 13 publications

References 20 publications

Assessment of the stallion sperm acrosome in two different extenders: Spermac stain in comparison with PNA/PSA/PI triple‐staining

Assessment of the stallion sperm acrosome in two different extenders: Spermac stain in comparison with PNA/PSA/PI triple‐staining

Bull Sperm SWATH-MS-Based Proteomics Reveals Link between High Fertility and Energy Production, Motility Structures, and Sperm–Oocyte Interaction

Abnormal Centriolar Biomarker Ratios Correlate with Unexplained Bull Artificial Insemination Subfertility – a Pilot Study

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