ProAKAP4 protein marker: Towards a functional approach to male fertility

Carracedo, Sara; Briand-Amirat, Lamia; Dordas-Perpinyà, Marta; Yolanda, Ramos Escuredo; Romain, Delcombel; Sergeant, Nicolas; Delehedde, Maryse

doi:10.1016/j.anireprosci.2022.107074

Cited by 13 publications

(20 citation statements)

References 82 publications

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“…A commercial ELISA kit for the assessment of proAKAP4 concentrations (4MID® Kits, 4BioDx, Lille, France) is available for human and 10 animal species, including the domestic cat. However, although there are many publications about proAKAP4 for multiple species (reviewed by Carracedo et al., 2022), we did not find any publications on its use for feline spermatozoa.…”

Section: Introductioncontrasting

confidence: 73%

“…It contributes to the formation of the ribs and columns of the fibrous sheath and anchors the glycolytic enzymes that regulate sperm motility (Delehedde et al., 2019). The AKAP4 protein is expressed in a variety of animal species, from lizards to mammals (Carracedo et al., 2022), and its gene sequence is highly conserved (with an overall homology of 70%), highlighting its important role in the spermatozoon (Carracedo et al., 2022). Its essential contribution to sperm motility is further demonstrated in AKAP4 knockout male mice, which are infertile due to an abnormal flagellar structure that causes markedly reduced sperm motility and a lack of progressive motility (Miki et al., 2002).…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of a commercial proAKAP4 kit for the assessment of fresh and frozen–thawed feline spermatozoa

Prochowska,

Eberhardt,

Niżański

2024

Reprod Domestic Animals

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ProAKAP4 is gaining increasing attention as a potential marker of semen quality in many species, but while there is a commercial kit for assessing proAKAP4 in the domestic cat, there are no publications about its use. The aim of this study was to evaluate the commercial proAKAP4 kit – Cat 4MID® Kit (SPQI – 4BioDx, Lille, France) for the assessment of feline semen. Semen was collected from 54 male cats by urethral catheterization. After a basic semen evaluation (subjective motility, CASA, viability, morphology), proAKAP4 levels in each sample were assessed using the Cat 4MID® Kit according to the manufacturer's protocol or with some modifications related to incubation time, sample storage conditions and number of spermatozoa used. Finally, the Spearman correlation of proAKAP4 concentration and sperm motility parameters was calculated. The most reliable results (acceptable intraassay coefficient of variation) were obtained with an optimized protocol of overnight incubation and isolation of proAKAP4 protein from 1 × 106 spermatozoa stored at −80°C. For fresh semen, there were no significant correlations between proAKAP4 concentration and sperm motility parameters, despite a strong correlation between motility parameters and sperm viability and morphology. A predominant effect of other sperm parameters and highly variable performance of lysis buffer question the usefulness of Cat 4MID® Kit for the assessment of feline semen. For frozen–thawed semen, there was a moderate, negative correlation between proAKAP4 concentration and two CASA parameters, VAP and VSL. As there were no correlations between proAKAP4 concentration in fresh semen and motility parameters in cryopreserved samples, proAKAP4 cannot be used as freezability marker in cats. More studies are needed to establish potential correlation with long‐lasting motility.

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Section: Introductioncontrasting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

Evaluation of a commercial proAKAP4 kit for the assessment of fresh and frozen–thawed feline spermatozoa

Prochowska,

Eberhardt,

Niżański

2024

Reprod Domestic Animals

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“…AKAP4 structural protein (an A-kinase anchoring protein which binds to PKA regulatory proteins) is the most abundant constitutive protein in the fibrous sheath of mammalian sperm and have been shown to be a key molecule involved in the regulation of flagellar motion and function [ 37 , 166 , 167 , 168 ]. Furthermore, AKAP4 interacts with many protein partners essential in regulation of sperm motility and hyperactivation including kinases, phosphatases, and glycolytic enzymes [ 80 , 169 , 170 ]. Reduced expression of AKAP4 has been associated with reduced motility and infertility [ 168 , 171 , 172 ].…”

Section: Methods Of Motility Assessmentmentioning

confidence: 99%

“…As such, proAKAP4, the precursor to AKAP4, has been identified as a promising marker for assessing sperm motility and quality in many mammalian species including the ram [ 169 ]. Concentrations of proAKAP4 can be determined using molecular assays like the 4MID Kit (4BioDx, Lille, France).…”

Section: Methods Of Motility Assessmentmentioning

confidence: 99%

“…Concentrations of proAKAP4 can be determined using molecular assays like the 4MID Kit (4BioDx, Lille, France). This kit is a quantitative immunoassay based on the sandwich ELISA which includes two monoclonal antibodies specific to the proAKAP4 protein [ 169 ]. The effectives of such a molecular approach in assessing motility and predicting long term quality of ram sperm has been reviewed by Carracedo, et al [ 169 ].…”

Section: Methods Of Motility Assessmentmentioning

confidence: 99%

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Motility Assessment of Ram Spermatozoa

Hoek

Rickard

Graaf

2022

Biology

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For successful fertilisation to occur, spermatozoa need to successfully migrate through the female reproductive tract and penetrate the oocyte. Predictably, poor sperm motility has been associated with low rates of fertilisation in many mammalian species, including the ram. As such, motility is one of the most important parameters used for in vitro evaluation of ram sperm quality and function. This review aims to outline the mechanical and energetic processes which underpin sperm motility, describe changes in motility which occur as a result of differences in sperm structure and the surrounding microenvironment, and assess the effectiveness of the various methods used to assess sperm motility in rams. Methods of subjective motility estimation are convenient, inexpensive methods widely used in the livestock industries, however, the subjective nature of these methods can make them unreliable. Computer-assisted sperm analysis (CASA) technology accurately and objectively measures sperm motility via two-dimensional tracing of sperm head motion, making it a popular method for sperm quality assurance in domesticated animal production laboratories. Newly developed methods of motility assessment including flagellar tracing, three-dimensional sperm tracing, in vivo motility assessment, and molecular assays which quantify motility-associated biomarkers, enable analysis of a new range of sperm motion parameters with the potential to reveal new mechanistic insights and improve ram semen assessment. Experimental application of these technologies is required to fully understand their potential to improve semen quality assessment and prediction of reproductive success in ovine artificial breeding programs.

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