Expression patterns of the activator type of cAMP-responsive element modulator in testicular germ cells of Japanese Black bulls

Noda, Taichi; Minami, Kenta; Kojima, Aya; Mizuno, Yohei; Isono, Ayane; Sakase, Mitsuhiro; Fukushima, Moriyuki; Harayama, Hiroshi

doi:10.1016/j.theriogenology.2014.01.014

Cited by 7 publications

(2 citation statements)

References 35 publications

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“…Therefore, variation in the level of AQP proteins in each ejaculate was considered to have been determined during spermatogenesis. Individual differences in mRNA and in amounts of protein expressed by some genes have been reported in the testis and spermatozoa [ 40 , 41 ]. Some conditions during spermatogenesis in each bull may influence the variation in the amount of AQPs among ejaculates, but further studies are required.…”

Section: Discussionmentioning

confidence: 99%

Expression and localization of aquaporins 3 and 7 in bull spermatozoa and their relevance to sperm motility after cryopreservation

Fujii

Hirayama

Fukuda

et al. 2018

Journal of Reproduction and Development

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Artificial insemination with cryopreserved semen is a well-developed technique commonly used for controlled reproduction in cattle. However, despite current technical advances, cryopreservation continues to damage bull spermatozoa, resulting in a loss of approximately 30 to 50% of viable spermatozoa post thawing. To further improve the efficiency of cryopreservation of bull spermatozoa, understanding the molecular mechanisms underlying the cryobiological properties that affect cryoinjuries during cryopreservation process of bull spermatozoa is required. In this study, we examined the expression and localization of aquaporin (AQP) 3 and AQP7 in fresh, cooled, and frozen-thawed bull spermatozoa. Furthermore, we investigated the relevance of AQP3 and AQP7 to motility and to membrane integrity in frozen-thawed bull spermatozoa. Western blotting against AQP3 and AQP7 in bull spermatozoa revealed bands with molecular weights of approximately 42 kDa and 53 kDa, respectively. In immunocytochemistry analyses, immunostaining of AQP3 was clearly observed in the principal piece of the sperm tail. Two immunostaining patterns were observed for AQP7 ―pattern 1: diffuse staining in head and entire tail, and pattern 2: diffuse staining in head and clear staining in mid-piece. Cooling and freeze-thawing did not affect the localization pattern of AQP7 and the relative abundances of AQP3 and AQP7 evaluated by Western blotting. Furthermore, we demonstrated that the relative abundances of AQP3 and AQP7 varied among ejaculates, and they were positively related to sperm motility, particularly sperm velocity, post freeze-thawing. Our findings suggest that AQP3 and AQP7 are possibly involved in the tolerance to freeze-thawing in bull spermatozoa, particularly in the sperm’s tail.

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Section: Discussionmentioning

confidence: 99%

Expression and localization of aquaporins 3 and 7 in bull spermatozoa and their relevance to sperm motility after cryopreservation

Fujii

Hirayama

Fukuda

et al. 2018

Journal of Reproduction and Development

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“…S9) were punctured with a 26‐gauge disposable needle and subsequently given gentle pressure to effuse the contents for collection with a micropipette. Cauda epididymal fluid was collected by gentle retrograde flushing with PBS after insertion of a 22‐gauge needle attached to a syringe (Terumo) into the ductus deferens (Noda et al, ). Samples from the testis and epididymis were immediately used for indirect immunofluorescence.…”

Section: Methodsmentioning

confidence: 99%

Distinct segment‐specific functions of calyculin A‐sensitive protein phosphatases in the regulation of cAMP‐triggered events in ejaculated bull spermatozoa

Mizuno

Isono

Kojima

et al. 2015

Molecular Reproduction Devel

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Livestock spermatozoa possess more tenacious suppressors of cAMP-triggered events-including capacitation-associated changes-than laboratory animal spermatozoa, leading to flagellar hyperactivation. In order to identify the suppressors, we examined effects of an inhibitor of serine/threonine protein phosphatases (calyculin A) on cAMP-triggered changes in the protein phosphorylation state, and subsequent occurrence of hyperactivation and acrosome reaction in ejaculated bull spermatozoa. Ejaculated spermatozoa were incubated in cAMP-supplemented medium, then assessed for motility, acrosome morphology, and phosphorylated protein localization. The addition of calyculin A greatly enhanced cAMP-triggered protein phosphorylation at serine/threonine and tyrosine residues in the connecting piece and induction of flagellar hyperactivation. Most hyperactivated spermatozoa exhibited extremely asymmetrical bends at the middle piece, which produced intensive twisting or figure-eight movements. In the sperm head, however, cAMP-triggered dephosphorylation of serine/threonine-phosphorylated proteins and subsequent acrosome reaction were abolished by the addition of calyculin A. Based on these results, we suggest that calyculin A-sensitive protein phosphatases in the connecting piece are suppressors of cAMP-triggered events leading to hyperactivation. By contrast, similar protein phosphatases in the sperm head accelerate cAMP-triggered events leading to the acrosome reaction. These findings are consistent with the indication that calyculin A-sensitive protein phosphatases have distinct functions in the regulation of cAMP-triggered events in different regions of ejaculated bull spermatozoa.

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Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction

Ogura

Takagishi

Harayama

2016

Animal Reproduction Science

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Expression patterns of the activator type of cAMP-responsive element modulator in testicular germ cells of Japanese Black bulls

Cited by 7 publications

References 35 publications

Expression and localization of aquaporins 3 and 7 in bull spermatozoa and their relevance to sperm motility after cryopreservation

Expression and localization of aquaporins 3 and 7 in bull spermatozoa and their relevance to sperm motility after cryopreservation

Distinct segment‐specific functions of calyculin A‐sensitive protein phosphatases in the regulation of cAMP‐triggered events in ejaculated bull spermatozoa

Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction

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