Calpain inhibition prevents flotillin re-ordering and Src family activation during capacitation

Maldonado-García, Deneb; Salgado‐Lucio, Monica L.; Roa-Espitia, Ana Lilia; Reyes-Miguel, Tania; Hernández‐González, Enrique O.

doi:10.1007/s00441-017-2591-2

Cited by 8 publications

(12 citation statements)

References 89 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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%

“…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. 12,23 These patterns are shown in Figure 2B. After 60 minutes of capacitation, patterns B and AR increased significantly in comparison with noncapacitated spermatozoa (P = .001 and P = .002, respectively; Figure 2B).…”

Section: Rac1 Is Involved In Capacitation and Armentioning

confidence: 89%

“…These assays were performed following the procedure described by Maldonado‐Garcia et al Noncapacitated and capacitated spermatozoa, treated or no with NSC23766 during 1 hour, were fixed with a mixture of formaldehyde (3% vol/vol) and glutaraldehyde (0.2% vol/vol) diluted in phosphate‐buffered saline (PBS). Spermatozoa were pelleted by centrifugation (600 g for 3 minutes), resuspended and incubated in NH 4 Cl (50 mM) for 10 minutes, rinsed twice with PBS, and once with bidistilled water.…”

Section: Methodsmentioning

confidence: 99%

“…Fluorescence images were registered using an Olympus BX5 fluorescence microscope. CTC staining patterns, 100 spermatozoa per sample were classified as Maldonado‐Garciía et al: ‘F,’ the acrosome region displays a faint fluorescence, feature of spermatozoa noncapacitated and with acrosome‐intact; “B,” a band along the equatorial segment and the acrosomal region exhibit a bright fluorescence, attribute of capacitated, and with acrosome‐intact spermatozoa; and “AR,” fluorescence in the postacrosomal region is distinctive of physiologically capacitated acrosome‐reacted spermatozoa. The status of the acrosomal cap on each spermatozoon was verified under phase‐contrast illumination.…”

Section: Methodsmentioning

confidence: 99%

“…We followed the procedure characterized by Ward and Storey 25 and adapted for guinea pig spermatozoa. 23 In a light-shielded container were mixed 45 μL of spermatozoa and the same volume of CTC solution (750 mM CTC diluted in 130 mM NaCl, 5 mM cysteine, and 20 mM Tris, at a pH of 7.8) and placed in a water bath at 37°C for 20 seconds. Then, the spermatozoa were fixed, adding glutaraldehyde (0.75 µL, final concentration 0.1%) diluted in Tris (0.5 mM, pH 7.4) at room temperature.…”

Section: Ctc Fluorescence Proceduresmentioning

confidence: 99%

See 4 more Smart Citations

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

View full text Add to dashboard Cite

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.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Rac1 Is Involved In Capacitation and Armentioning

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Ctc Fluorescence Proceduresmentioning

confidence: 99%

See 3 more Smart Citations

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

View full text Add to dashboard Cite

show abstract

Perfluorooctane sulfonate and perfluorooctanoic acid induce plasma membrane dysfunction in boar spermatozoa during in vitro capacitation

Ortiz-Sánchez

Roa-Espitia

Fierro

et al. 2022

Reproductive Toxicology

View full text Add to dashboard Cite

Guinea pig spermatozoa adhesion to an immobilized fibronectin matrix alters their physiology and increases their survival

Jorge-Cruz,

Roa-Espitia,

Hernández-González

2024

Biology of Reproduction

View full text Add to dashboard Cite

Isthmus is the region of the oviduct considered a reservoir for spermatozoa, where they are retained and then released synchronously with ovulation. Integrins mediate this interaction, and it is suggested that they regulate the viability and capacitation of spermatozoa. Spermatozoa retained in the oviductal epithelial cells show specific characteristics: normal morphology, intact acrosome and plasma membrane, no DNA fragmentation, and low levels of intracellular Ca2+ and protein phosphorylation at Tyr. This work aimed to define spermatozoa’s ability to adhere to an immobilized fibronectin matrix and its effects on their viability and capacitation. We found that guinea pig spermatozoa showed a high affinity for adhering to an immobilized fibronectin matrix but not to those made up of type 1 collagen or laminin. This interaction was mediated by integrins that recognize the RGD domain. Spermatozoa adhered to an immobilized fibronectin matrix were maintained in a state of low capacitation: low levels of intracellular concentration of Ca2+, protein phosphorylation in Tyr and F-actin. Also, spermatozoa kept their plasma membrane and acrosome intact, flagellum beating, and showed low activation of caspases 3/7. The spermatozoa adhered to the immobilized fibronectin matrix, gradually detached, forming rosettes and did not undergo a spontaneous acrosomal reaction but were capable of experiencing a progesterone-induced acrosomal reaction. In conclusion, the adhesion of spermatozoa to an immobilized fibronectin matrix alters the physiology of the spermatozoa, keeping them in a steady state of capacitation, increasing their viability in a similar way to what was reported for spermatozoa adhered to oviductal epithelial cells.

show abstract

Calpain inhibition prevents flotillin re-ordering and Src family activation during capacitation

Cited by 8 publications

References 89 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

Perfluorooctane sulfonate and perfluorooctanoic acid induce plasma membrane dysfunction in boar spermatozoa during in vitro capacitation

Guinea pig spermatozoa adhesion to an immobilized fibronectin matrix alters their physiology and increases their survival

Contact Info

Product

Resources

About