ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm

Riesco, Marta F.; Anel-López, Luis; Neila-Montero, Marta; Palacin-Martinez, Cristina; Montes-Garrido, Rafael; Álvarez, M.; Paz, P. de; Anel, L.

doi:10.3390/biom10071046

Cited by 38 publications

(49 citation statements)

References 70 publications

(110 reference statements)

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“…Jumeau et al (2018) stated that there was a positive correlation between the proAKAP4 level and the sperm motility of human spermatozoa separated from seminal plasma, detected using the SDS‐PAGE and protein immunoblot methods. Previous studies performed in stallion, ram, dog and bull frozen‐thawed semen samples reported a positive correlation between the proAKAP4 levels determined by ELISA and CASA techniques performed at hour 0 (post‐thawed) in terms of total motility, progressive motility, BCF, LIN, VCL and VSL (Blommaert et al., 2019; Le Couazer et al., 2019; Riesco et al., 2020; Ruelle et al, 2019). In addition to the CASA results, Ruelle et al (2019) stated that the correlation was associated with the success rates of approximately 200,000 AI practices to support their hypothesis.…”

Section: Discussionmentioning

confidence: 92%

Quality assessment of frozen bull semen with the precursor A‐kinase anchor protein 4 biomarker

Baştan

Akçay

2021

Andrologia

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In this study, the quality of frozen bull semen was evaluated with the proAKAP4 level test. Sixty straws of frozen bull semen from various batches (n = 30) belonging to six bulls were used in the current study. The frozen bull semen samples were analysed in terms of proAKAP4 levels, sperm morphology and sperm movement parameters at hour 0 and hour 3 after thawing. The semen samples were divided into three groups according to the proAKAP4 levels: low concentration (<25 ng/10x106 spermatozoa), moderate concentration (25 to 39 ng/10x106 spermatozoa) and high concentration (≥40 ng/10x106 spermatozoa). A positive correlation was found between the proAKAP4 level and total motility (TM3), progressive motility (PM3), VSL3 and VCL3 values obtained after the third‐hour thermoresistance test (p < .05). There was a negative correlation between the percentage of sperm abnormal tail and the proAKAP4 level (p < .01). In addition, it was observed that the semen samples with proAKAP4 concentrations of 25 ng/106 spermatozoa and higher preserved the TM3 and PM3 motility characteristics. In conclusion, the proAKAP4 has the potential to become a biomarker protein to evaluate in the quality analysis of frozen‐thawed semen.

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Section: Discussionmentioning

confidence: 92%

Quality assessment of frozen bull semen with the precursor A‐kinase anchor protein 4 biomarker

Baştan

Akçay

2021

Andrologia

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“…Probably this negative effect is due to some deleterious effects at the structural level since the sperm metabolism was not affected by the gentamicin respect to the C samples. A recent study carried out by Riesco et al ( 32 ) demonstrated a novel sperm protein (ProAKAP4) as a promising diagnostic parameter of sperm quality in ram sperm, so long correlated to sperm motility. This protein is the most expressed protein of the sperm fibrous sheath being an important part of the flagellum ( 33 , 34 ).…”

Section: Discussionmentioning

confidence: 99%

Comparing the Effect of Different Antibiotics in Frozen-Thawed Ram Sperm: Is It Possible to Avoid Their Addition?

et al. 2021

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It is crucial to perform a deep study about the most extensively used antibiotics in sperm extenders. Most of the protocols and concentrations used in ram are direct extrapolations from other species. It is important to establish species-specific antibiotic treatments to optimize their use and if it is possible to reduce the quantity. Previews studies have assessed some aspects of sperm quality in vitro, but this study aimed to go further and assess the effect of three different antibiotic treatments, which are the most extensively used, not only in sperm quality or assessing the inhibitory effect on bacterial growth but also assessing these important parameters of productivity such as fertility, prolificacy, fecundity, and sex-ratio during a freeze-thaw process. Gentamicyn (G) treatment showed the worst results, not only concerning sperm quality but also in the reproductive trials exhibiting a toxical effect at the experiment concentration, and being the most powerful inhibiting bacterial growth. For its part, Lincomicyn-spectinomycin (LS) showed similar results inhibiting bacterial growth but it did not show a detrimental effect either in sperm quality or in reproductive parameters. Penicillin-streptomycin (PS) showed good results in the sperm quality and in the reproductive in vivo trials, but it showed a very poor effect inhibiting bacterial growth probably due to some kind of antibiotic resistance. According to our results, there is not a significant positive relationship between the higher bacterial inhibitory activity of LS and PS samples, and the sperm quality respect Control samples (without antibiotics). In the case of G, which exhibited the most effective as antibacterial, we observed a toxic effect on sperm quality that could be translated on productivity parameters. Our results suggest that the bacterial contamination control in frozen-thawed semen may be possible without the use of antibiotics, although the effects of longer periods of cooling storage and different temperatures of storage need to be further investigated for animal semen. At this point, a reflection about a drastic reduction in the use of antibiotic treatments in sperm cryopreservation is mandatory, since freezing conditions could keep sperm doses contamination within the levels recommended by regulatory health agencies.

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“…Sperm samples of different experimental groups were diluted in phosphate-buffered saline (PBS) medium to obtain a total of 2 × 10 6 of spermatozoa per sample; these samples were centrifuged at 500× g for 10 min at RT. The flow cytometry staining protocol has been previously described by Riesco et al, 2020 [ 29 ]. Briefly, the supernatant was discarded and the sperm pellet was incubated at RT for 30 min in the dark with 96 μL of Zombie Violet™ (membrane integrity probe) (1:1000 final dilution in PBS, Biolegend, Madrid, Spain), 2 μL of CellEvent™ Caspase-3/7 (apoptosis marker) (4 μM final concentration in PBS, Thermo Fisher, Madrid, Spain) and 2 μL of CellROX ® (ROS content labeling) (5 μM final concentration in PBS, Thermo Fisher, Madrid, Spain).…”

Section: Methodsmentioning

confidence: 99%

Multiparametric Study of Antioxidant Effect on Ram Sperm Cryopreservation—From Field Trials to Research Bench

Riesco

Álvarez

Anel-López

et al. 2021

Animals

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The optimization of sperm cryopreservation protocols in ram is a feasible tool to reinforce artificial insemination technologies considering the desirable application of sperm by vaginal/cervical or transcervical deposition. Cryopreservation provokes different types of damage on spermatozoa and many of these detrimental effects are triggered by redox deregulation. For this reason, the antioxidant supplementation in sperm cryopreservation protocols to decrease reactive oxygen species (ROS) levels and to equilibrate redox status has been widely employed in different species. Despite this, more fertility trials are necessary to provide the definitive tool to ensure the antioxidant effectiveness on sperm quality. For this reason, in this work, we performed a multiparametric analysis of some previously tested antioxidants (crocin, GSH and Trolox) on ram sperm cryopreservation from field trials to sperm quality analyses focused on new strategies to measure redox balance. Attending to fertility trial, Trolox supplementation registered an improvement concerning to fertility (when we considered high fertility males) and multiple lambing frequency and other complementary and descriptive data related to lambing performance such as prolificacy and fecundity. This positive effect was more evident in multiple lambing frequency when we considered low fertility males than in global male analysis. In vitro analyses of sperm quality confirmed in vivo trials registering a positive effect on sperm viability and redox balance. In this study, we provided the definitive evidence that the role of trolox on redox balance maintenance has a direct effect on fertility parameters, such as prolificacy. The effectiveness of antioxidant treatments was tested, for the first time in ovine species, using an integrative and multiparametric approach combining in vivo and in vitro analyses and novel approaches, such as RedoxSYS. These types of strategies should be applied to improve sperm conservation methods and optimize AI technologies upgrading the correlation between in vitro and in vivo analyses.

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ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm

Cited by 38 publications

References 70 publications

Quality assessment of frozen bull semen with the precursor A‐kinase anchor protein 4 biomarker

Quality assessment of frozen bull semen with the precursor A‐kinase anchor protein 4 biomarker

Comparing the Effect of Different Antibiotics in Frozen-Thawed Ram Sperm: Is It Possible to Avoid Their Addition?

Multiparametric Study of Antioxidant Effect on Ram Sperm Cryopreservation—From Field Trials to Research Bench

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