Focal adhesion kinase is required for actin polymerization and remodeling of the cytoskeleton during sperm capacitation

Roa-Espitia, Ana Lilia; Hernández-Rendón, Eva R.; Baltiérrez-Hoyos, Rafael; Muñoz-Gotera, Rafaela; Cote-Vélez, Antonieta; Jiménez, Irma; González‐Márquez, Humberto; Hernández‐González, Enrique O.

doi:10.1242/bio.017558

Cited by 46 publications

(61 citation statements)

References 70 publications

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“…The cellular distribution of β1‐integrin was analyzed in capacitated spermatozoa in the absence or presence of NSC23766 (100 μM). In noncapacitated and capacitated spermatozoa, β1‐integrin was located in the apical region of the acrosome and the midpiece (Non‐Cap and Cap; Figure ), as has already been reported . These results indicate that β1‐integrin does not change its location during capacitation (Figure ).…”

Section: Resultssupporting

confidence: 81%

“…Capacitated spermatozoa presented an increase in fluorescence, mainly in the apical region of the acrosome, the flagellum and the equatorial segment (FAP2; Figure A). These two patterns are typically observed during the capacitation of guinea pig spermatozoa . Spermatozoa capacitated in the presence of NSC23766 presented a third pattern of increased staining, but unlike FAP1 and FAP2, the increase occurred throughout the acrosome, except for the apical region (FA3P; Figure A).…”

Section: Resultsmentioning

confidence: 80%

“…It is known, that actin cytoskeleton is remodeled in the apical region of the acrosome during capacitation, forming a barrier that prevents the premature AR . A similar remodeling of the actin cytoskeleton has been described for capacitated spermatozoa of bovine, boar, guinea pig, and ram; an essential feature of these spermatozoa is their spatula‐shaped head …”

Section: Introductionmentioning

confidence: 61%

“…Spermatozoa were capacitated (60 minutes) in the absence and presence of NSC23766 (100 μM). In other studies, we have defined the different fluorescent patterns observed when spermatozoa are stained with CTC: pattern F for noncapacitated spermatozoa, pattern B for capacitated spermatozoa, and AR pattern for sperm that have experienced AR . These patterns are shown in Figure B.…”

Section: Resultsmentioning

confidence: 99%

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Rac1 is necessary for capacitation and acrosome reaction in guinea pig spermatozoa

Ramírez‐Ramírez

Salgado‐Lucio²,

Roa-Espitia³

et al. 2019

J of Cellular Biochemistry

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Actin cytoskeleton remodeling is a critical process for the acquisition of fertilizing capacity by spermatozoa during capacitation. However, the molecular mechanism that regulates this process has not been fully elucidated. In somatic cells, Ras‐related C3 botulinum toxin substrate 1 protein (Rac1) promotes the polymerization of actin by participating in the modeling of two structures: lamellipodia and adhesion complexes linked with the plasma membrane. Rac1 is expressed in mammalian spermatozoa; however, the role of Rac1 in sperm physiology is unknown. This study aimed to elucidate the participation of Rac1 in capacitation and acrosome reaction (AR). Rac1 was found to be dispersed throughout the acrosome and without changes in the middle piece. After 60 minutes of capacitation, Rac1 was found in the apical region of the acrosome only, which concurred with an increase in Rac1‐GTP. Rac1 inhibition prevented such changes. In the middle piece, Rac1 localization remained unchanged. Besides, Rac1 inhibition blocked capacitation and AR. The present study demonstrates that Rac1 participates only in the actin cytoskeleton remodeling that occurs in the acrosomal apical region during capacitation, a region where a large amount of actin is polymerized and shaped in a diadem‐like structure. Our data also show that this actin cytoskeleton organized by Rac1 interacts with filamin‐1, and such interaction was blocked by the inhibition of Rac1, which led to a different organization of the actin cytoskeleton. All these outcomes imply that the formation of an F‐actin cytoskeleton in the acrosomal apical region is a necessary event for capacitation and AR, and which is Rac1 driven.

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Section: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 61%

Section: Resultsmentioning

confidence: 99%

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Rac1 is necessary for capacitation and acrosome reaction in guinea pig spermatozoa

Ramírez‐Ramírez

Salgado‐Lucio²,

Roa-Espitia³

et al. 2019

J of Cellular Biochemistry

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“…Alpha-actinin-4 interacts with the focal-adhesive complex, ensuring it is connected to the actin cytoskeleton [55]. This connection supports the activity of the focaladhesive kinase, which provides the acrosome the integrity necessary for capacitation [56]. Consequently, changes in the alpha-actinin-4 content in spermatozoa can have significant consequences for their fertilization ability.…”

Section: Mrna and Protein Expression Levels Of Cytoskeletal Genesmentioning

confidence: 85%

Cytoskeleton Structure in Mouse Sperm and Testes After 30 Days of Hindlimb Unloading and 12 Hours of Recovery

Usik,

Ogneva

2018

Cell Physiol Biochem

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Background/Aims: Changes in the external mechanical field result in cytoskeleton reorganization and the formation of adaptive patterns in different types of cells, including somatic cells and sex cells. The aim of this research was to study the protein and mRNA content of cytoskeletal and sperm-specific genes in the sperm and testis cells of mice. Methods: Mice were subjected to 30 days of antiorthostatic suspension to simulate weightlessness, followed by 12 h of recovery, while receiving essential phospholipids at a dosage of 500 mg/kg/day (30HSE and 30HSE+12h groups) or a similar dosage of a placebo (30HS and 30HS+12h groups). Accordingly, reference groups (CE group and C group) were formed. The total number and the percentage of motile spermatozoa were calculated using a Makler chamber. To analyze the number of viable spermatozoa and the permeability of their membranes, eosin staining was used as well as Diff-Quick for a morphological evaluation. Relative protein and mRNA content was estimated in a western blot and quantitative PCR assay, respectively. Results: The relative protein expression levels of actin (beta and gamma) and two alpha-actinin isoforms (1 and 4) remained constant in the sperm of all study groups, except for the 30HS+12h group, where the alpha-actinin-4 level was 13% higher than in the reference group (p < 0.1). In the testis cells, the relative actin isoform content was equivalent to that in the spermatozoa. However, in the testis cells, the ACTN1 mRNA content was 17% higher in the 30HS group than in the C group (p < 0.05), and decreased after 12 h of recovery. In contrast, the ACTN4 mRNA content was 20% lower in the 30HS group than in the reference group (p < 0.05) and increased after the 12-h recovery period. At the same time, in the group administered the essential phospholipids, the relative ACTN1 and ACTN4 mRNA content did not differ from those of the reference group. The relative beta-tubulin content was similar in the reference C group and the reference CE group, which was administered the essential phospholipids. In the 30HS and 30HS+12h groups, the beta-tubulin content decreased by 19% and 22% (p < 0.05), respectively, and they also decreased in the groups administered the essential phospholipids (30HSE and 30HSE+12h groups, by 27% and 33%, respectively, p < 0.05). In the testis tissue, the relative tubulin content did not change in any of the experimental groups. At the same time, the relative mRNA content of the genes encoding the studied cytoskeletal proteins increased, which may indicate the protein content was regulated mainly at the translational level. Conclusion: The spermogram parameters and the content of the sperm-specific proteins and the associated mRNAs revealed a decrease in the number of mature spermatozoa in mice suspended under conditions of weightlessness. Moreover, the decrease was prevented by the administration of essential phospholipids.

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TMEM16A inhibition impedes capacitation and acquisition of hyperactivated motility in guinea pig sperm

Cordero-Martínez

Reyes-Miguel

Rodrı́guez-Páez

et al. 2018

J of Cellular Biochemistry

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Ca -activated Cl channels (CaCCs) are anionic channels that regulate many important physiological functions associated with chloride and calcium flux in some somatic cells. The molecular identity of CaCCs was revealed to be TMEM16A and TMEM16B (also known as Anoctamin or ANO1 and ANO2, respectively) in all eukaryotes. A recent study suggests the presence of TMEM16A in human sperm and a relationship with the rhZP-induced acrosome reaction. However, to the best of our knowledge, little is known about the role of TMEM16A in other spermatic processes such as capacitation or motility. In this study, we evaluated the effects of two TMEM16A antagonists on capacitation, acrosome reaction, and motility in guinea pig sperm; these antagonists were T16Ainh-A01, belonging to a second generation of potent antagonists of TMEM16A, and niflumic acid (NFA), a well-known antagonist of TMEM16A (CaCCs). First of all, we confirmed that the absence of Cl in the capacitation medium changes motility parameters, capacitation, and the progesterone-induced acrosome reaction. Using a specific antibody, TMEM16A was found as a protein band of ∼120 kDa, which localization was in the apical crest of the acrosome and the middle piece of the flagellum. Inhibition of TMEM16A by T16Ainh-A01 affected sperm physiology by reducing capacitation, blocking the progesterone-induced acrosome reaction under optimal capacitation conditions, inhibiting progressive motility, and the acquisition of hyperactivated motility, diminishing [Ca ]i, and increasing [Cl ]i. These changes in sperm kinematic parameters provide new evidence of the important role played by TMEM16A in the production of sperm capable of fertilizing oocytes.

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Focal adhesion kinase is required for actin polymerization and remodeling of the cytoskeleton during sperm capacitation

Cited by 46 publications

References 70 publications

Rac1 is necessary for capacitation and acrosome reaction in guinea pig spermatozoa

Rac1 is necessary for capacitation and acrosome reaction in guinea pig spermatozoa

Cytoskeleton Structure in Mouse Sperm and Testes After 30 Days of Hindlimb Unloading and 12 Hours of Recovery

TMEM16A inhibition impedes capacitation and acquisition of hyperactivated motility in guinea pig sperm

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