The motility-based swim-up technique separates bull sperm based on differences in metabolic rates and tail length

Magdanz, Veronika; Boryshpolets, Sergii; Ridzewski, Clara; Eckel, Barbara; Reinhardt, Klaus

doi:10.1371/journal.pone.0223576

Cited by 49 publications

(40 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Swim up was done according to the standardised protocol [ 31 ] with some modifications. After the initial wash, the sperm pellet was re-suspended and washed 1× in 300 μL of Sperm-TALP medium, which was followed by a brief centrifugation (3 min, 500× g ).…”

Section: Methodsmentioning

confidence: 99%

Mitochondria Content and Activity Are Crucial Parameters for Bull Sperm Quality Evaluation

et al. 2021

View full text Add to dashboard Cite

Standard sperm evaluation parameters do not enable predicting their ability to survive cryopreservation. Mitochondria are highly prone to suffer injuries during freezing, and any abnormalities in their morphology or function are reflected by a decline of sperm quality. Our work focused on describing a link between the number and the activity of mitochondria, with an aim to validate its applicability as a biomarker of bovine sperm quality. Cryopreserved sperm collected from bulls with high (group 1) and low (group 2) semen quality was separated by swim up. The spermatozoa of group 1 overall retained more mitochondria (MitoTrackerGreen) and mtDNA copies, irrespective of the fraction. Regardless of the initial ejaculate quality, the motile sperm contained significantly more mitochondria and mtDNA copies. The same trend was observed for mitochondrial membrane potential (ΔΨm, JC-1), where motile sperm displayed high ΔΨm. These results stay in agreement with transcript-level evaluation (real-time polymerase chain reaction, PCR) of antioxidant enzymes (PRDX1, SOD1, GSS), which protect cells from the reactive oxygen species. An overall higher level of glutathione synthetase (GSS) mRNA was noted in group 1 bulls, suggesting higher ability to counteract free radicals. No differences were noted between basal oxygen consumption rate (OCR) (Seahorse XF Agilent) and ATP-linked respiration for group 1 and 2 bulls. In conclusion, mitochondrial content and activity may be used as reliable markers for bovine sperm quality evaluation.

show abstract

Section: Methodsmentioning

confidence: 99%

Mitochondria Content and Activity Are Crucial Parameters for Bull Sperm Quality Evaluation

et al. 2021

View full text Add to dashboard Cite

show abstract

“…The use of a swim-up method to select sperm, instead of protocols requiring density gradient centrifugations, is a clear example of such approximation. This procedure is routinely used in bovine and human seminal samples to isolate spermatozoa with high rates of normal morphology and motility rates and decreased DNA fragmentation [ 35 , 36 ]. In pigs, swim-up is an appropriate experimental system for sperm purification providing similarities with spermatozoa selected in vivo [ 12 ].…”

Section: Discussionmentioning

confidence: 99%

Replacement of Albumin by Preovulatory Oviductal Fluid in Swim-Up Sperm Preparation Method Modifies Boar Sperm Parameters and Improves In Vitro Penetration of Oocytes

Navarro-Serna

París-Oller

Šimoník

et al. 2021

Animals

View full text Add to dashboard Cite

More suitable and efficient methods to protect gametes from external harmful effects during in vitro handling can be achieved by adding preovulatory porcine oviductal fluid (pOF) to in vitro culture media. The objective of this study was to assess the swim-up procedure’s suitability as a sperm selection method using a medium supplemented with 1mg/mL BSA, 1% preovulatory pOF (v/v), 1% v/v pOF plus 1mg/mL BSA, and 5mg/mL BSA. After selection, various sperm parameters were studied, such as sperm recovery rate, sperm morphology, motility (by CASA), vitality, acrosome status and intracellular calcium (by flow cytometry) and ability to penetrate oocytes in vitro. Around 2% of sperm were recovered after swim-up, and the replacement of BSA by pOF showed a beneficial reduction of motility parameters calcium concentration, resulting in an increased penetration rate. The combination of albumin and oviductal fluid in the medium did not improve the sperm parameters results, whereas a high concentration of BSA increased sperm morphological abnormalities, motility, and acrosome damage, with a reduction of calcium concentration and penetration rate. In conclusion, the replacement of albumin by preovulatory oviductal fluid in the swim-up sperm preparation method modifies boar sperm parameters and improves the in vitro penetration of oocytes.

show abstract

“…The determination of sperm oxygen consumption using a Clark electrode has been used as a relatively simple method to evaluate the sperm mitochondrial function in several species such as stallion (42), human (43), boar (44), and ram (13). Oxygen consumption has often been considered a good indicator of the overall metabolic activity within cells, including spermatozoa (45,46), and it has also been correlated with fertility in bulls (17). In this study, a positive correlation between this parameter and reproductive results was found, but only in the B season.…”

Section: Discussionmentioning

confidence: 99%

Influence of Non-conventional Sperm Quality Parameters on Field Fertility in Ovine

et al. 2021

View full text Add to dashboard Cite

The prediction of the fertilizing ability of a seminal dose continues to be a primary aim in the field of artificial insemination (AI). To achieve this goal, in this study we have included the evaluation of some non-conventional sperm quality markers. A total of 3,906 ewes from 52 different farms were inseminated with 357 refrigerated seminal doses obtained from 45 mature Rasa Aragonesa rams. The same samples were used for sperm quality analysis including membrane integrity, capacitation status, oxygen consumption and apoptotic-like markers such as phosphatidylserine translocation (PS), plasmalemma disorganization/mitochondrial membrane potential, caspase activation and DNA damage. Seminal doses from the breeding (B) season presented higher percentages of intact membrane (IM), non permeant (NP) membrane with high mitochondrial membrane potential (ΔΨm) and IM without PS translocation spermatozoa than those from the non-breeding (NB) season. Therefore, we can conclude that there were less spermatozoa showing apoptotic-like features in the seminal doses from the B than the NB season, although these differences did not affect field fertility. Only the percentage of intact membrane, non-capacitated (IM-NC) spermatozoa showed a significant correlation with in vivo fertility (P = 0.005) and fecundity (P = 0.007) values obtained after cervical AI when all data were evaluated. When the data were sorted by season and distance to the farms where AI was performed, the correlation between the percentage of IM-NC spermatozoa and reproductive parameters increased in the NB season and progressively with remoteness from the farms. Some other sperm parameters, like NP with high ΔΨm, IM sperm without active caspases and DNA-intact spermatozoa, also showed significant correlations with the reproductive parameters in the sorted data. Moreover, the increment in both the percentage of IM-NC and DNA-intact spermatozoa would increase the probability of obtaining a fertility higher than the mean (>52%), as revealed by a multiple logistic regression analysis. In conclusion, we have identified two seminal markers—the percentage of intact membrane, non-capacitated spermatozoa, and DNA intact spermatozoa—which could be used as a test to discard males in AI programs, which is highly important from an economic point of view and can contribute to achieving satisfactory fertility rates.

show abstract

The motility-based swim-up technique separates bull sperm based on differences in metabolic rates and tail length

Cited by 49 publications

References 48 publications

Mitochondria Content and Activity Are Crucial Parameters for Bull Sperm Quality Evaluation

Mitochondria Content and Activity Are Crucial Parameters for Bull Sperm Quality Evaluation

Replacement of Albumin by Preovulatory Oviductal Fluid in Swim-Up Sperm Preparation Method Modifies Boar Sperm Parameters and Improves In Vitro Penetration of Oocytes

Influence of Non-conventional Sperm Quality Parameters on Field Fertility in Ovine

Contact Info

Product

Resources

About