Effects of calpain and Rho-kinase inhibitors on the acrosome reaction and motility of fowl spermatozoa in vitro

Ashizawa, Koji; Wishart, G. J.; Katayama, Seiichi; Takano, Daisuke; Maeda, Mineko; Arakawa, Emi; Tsuzuki, Yasuhiro

doi:10.1530/rep.1.00588

Cited by 13 publications

(9 citation statements)

References 64 publications

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“…Ca 2C influx from the extracellular medium to the cytosol through voltage channels is believed to be the first event that initiates the successive signaling pathways required for the acrosome exocytotic secretory response. The concomitant need of two factors, IPVL components and Ca 2C , for the in vitro induction of the chicken acrosome reaction has already been reported by different authors (Horrocks et al 2000, Ashizawa et al 2004, 2006a, 2006b). However, the present study showed that a small percentage of acrosome-reacted spermatozoa may be obtained with the presence of Ca 2C without IPVL in the BPSE and DMEM media, and in the three media studied after the addition of Ca 2C ionophore.…”

Section: Discussionmentioning

confidence: 71%

“…Okamura & Nishiyama (1978) described the ultrastructure of the acrosome reaction when spermatozoa are in contact to the IPVL of the ovulated oocyte. Horrocks et al (2000) suggested that the IPVL inducers of acrosome reaction could be N-linked oligosaccharides with terminal N-acetyl-glucosamine residues, while Ashizawa et al (2004Ashizawa et al ( , 2006aAshizawa et al ( , 2006b) started to describe the signaling pathways involved in the chicken acrosome reaction and Rabbani et al (2006Rabbani et al ( , 2007 suggested the involvement of sperm-associated bodies in the interaction of spermatozoa and IPVL. However, the understanding of the factors involved in the induction of the acrosome reaction is still very incomplete, despite its importance for the understanding of the process of fertilization itself and the need to counteract induction of the acrosome reaction when it occurs spontaneously after in vitro storage for example.…”

Section: Discussionmentioning

confidence: 99%

“…Fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA, Arachis hypogeae) (FITC-PNA) has been used to study acrosomal status (Horrocks et al 2000, Ashizawa et al 2004, 2006a, 2006b. Motility was evaluated by computer-assisted semen analyses (CASA) to measure any motility hyperactivation related to the initiation of acrosome reaction.…”

Section: Introductionmentioning

confidence: 99%

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A reappraisal of the factors involved in in vitro initiation of the acrosome reaction in chicken spermatozoa

Lemoine¹,

Grasseau²,

Brillard³

et al. 2008

Reproduction

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Chicken spermatozoa may remain in the female oviduct for a prolonged period before induction of the acrosome reaction on contact with the inner perivitelline layer (IPVL). By contrast, the acrosome reaction may be induced very rapidly in vitro in the presence of IPVL and Ca 2C. In the present study, we examined the extent to which the chicken acrosome reaction can be induced in media of various compositions in the presence or absence of IPVL and/or Ca 2C and other factors known to be efficient in mammals. We also compared the efficacy of perivitelline layer (PL) taken at various states of oocyte maturation in initiating the reaction. The acrosome reaction was induced in less than 5 min in the presence of Ca 2C and IPVL. Incubation of spermatozoa in different saline media (Beltsville poultry semen extender (BPSE); Dulbecco's modified eagle medium; NaCl-TES buffer) without IPVL showed a significant induction of acrosome reaction in BPSE supplemented with 5 mM Ca 2C and in the three media after supplementation with Ca 2C and Ca 2C ionophore A23187.By contrast, the acrosome reaction was never induced without Ca 2C. BSA, NaHCO 3 , and progesterone did not stimulate the acrosome reaction. Ca 2C plus PL taken at various physiological states (follicle IPVL, ovulated IPVL, oviposited IPVL, and/or outer perivitelline layer) strongly stimulated the acrosome reaction, the latest states being the most efficient. Although PL induced the acrosome reaction in the presence of extracellular Ca 2C, it was not possible to induce hyperactivation in chicken spermatozoa. Taken together, these results emphasize the central role of Ca 2C in the in vitro initiation of the acrosome reaction in chickens and show specific features of this induction in birds.

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Section: Discussionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A reappraisal of the factors involved in in vitro initiation of the acrosome reaction in chicken spermatozoa

Lemoine¹,

Grasseau²,

Brillard³

et al. 2008

Reproduction

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“…Ashizawa et al [17] showed that PKC may contribute to inhibition of the reaction. In addition, protein phosphatases type 1 and/or type 2A and protein phosphatase type 2B have been suggested to be involved in the regulation of the acrosome reaction in the chicken, because specific inhibition activities of these phosphatases significantly stimulate the reaction [17][18][19]. However, the potential involvement of kinases, such as MAPKs (MAPK3/1 and MAPK14), PKA, and PIK3, has not been investigated to date.…”

Section: Introductionmentioning

confidence: 99%

Potential Involvement of Several Signaling Pathways in Initiation of the Chicken Acrosome Reaction1

Lemoine¹,

Dupont²,

Guillory³

et al. 2009

Biology of Reproduction

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Avian sperm biology has demonstrated specific features in preparation for fertilization. For example, capacitationlike processes and motility hyperactivation do not exist in the form described in mammals. The present study investigated the potential involvement of several signaling pathways, including protein kinase A (PKA), phosphatidylinositol 3 kinase (PIK3), mitogen-activated protein kinase 3/1 (MAPK3/1), and MAPK14 in the chicken acrosome reaction (AR). The presence in chicken spermatozoa of key proteins involved in these signaling pathways (i.e., cAMP-responsive element-binding protein [CREB], AKT, MAPK1, and MAPK14 and their respective phosphorylated forms) was detected using immunoblotting and localized by immunocytochemistry, mainly in the heads. The potential involvement of these pathways in the AR induced by inner perivitelline layer (IPVL) and Ca(2+) was then examined using specific inhibitors and phosphorylation status measurements. The effects of the specific inhibitors on motility were also measured. Phosphorylations of AKT, CREB, and MAPK1, but not MAPK14, were increased at the time of AR. Phosphorylation of AKT was increased in the presence of IPVL alone, whereas both IPVL and Ca(2+) were needed to increase CREB and MAPK1 phosphorylations. Inhibition of the three corresponding pathways blocked the increase in phosphorylation and significantly decreased AR. Inhibitions of the PKA and MAPK1 pathways also significantly decreased motility, whereas MAPK14 and PIK3 inhibition had no effect on motility. Our results suggest that the AR could be mediated by activation of the PKA, PIK3, and MAPK1 pathways through a sequential action involving, successively, PIK3 and then PKA and MAPK1 activations.

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“…In presence of PVL the ability of sperm to perform the AR was not different (P>0.05) in fresh semen than thawed semen using BPSE diluent, however with Lake diluent the ability of sperm to perform the AR was lower (P<0.05) after thawed. Some studies have reported that cryopreservation impairs the ability of sperm to undergo an AR due to the reduced production of cAMP, lack of mobilization of Ca 2+ , or oxidative stress and changes in the proportion of cholesterol and phospholipids in the membranes before the AR (Ashizawa et al 2006). The percentage of sperm that underwent an AR was always higher (P<0.05) when they were induced with PVL than the percentage that underwent an AR without PVL.…”

Section: Resultsmentioning

confidence: 95%

Evaluation of two diluents for the storage of fresh and cryopreserved semen of Harris hawk (Parabuteo unicinctus)

Herrera

Calderón

Guzmán

et al. 2017

Austral j. vet. sci.

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ABSTRACT. Seminal storage, both fresh and cryopreserved, has contributed to the reproduction of wild birds in captivity, however, a method of predicting the fertilization capacity is needed. Indicators of the sperm acrosome reaction (AR) have been associated with its fertilization capacity. Therefore, the aim of this study was to evaluate the in vitro AR of fresh and cryopreserved Harris hawk sperm using Beltsville Poultry Semen Extender (BPSE) and Lake as diluents. 64 ejaculates were obtained and examined fresh and after thawing. The sperm basic evaluation for each ejaculate was made using eosin-nigrosin, while the ability of AR was assessed by coincubation with perivitelline layer (PVL). Sperm motility of fresh semen was higher (P<0.05) in fresh semen in BPSE (37.9±1.7) than in Lake (30.9±1.7) diluent. However, the motility decreased (P<0.05) in both diluents after thawing. For fresh semen, the percentage of sperm that underwent an AR without incubation with PVL was higher (P<0.05) with BPSE (14.1±1.7) than with Lake (6.8±2.5) diluent, however, AR was similar between tow diluents (P>0.05) after thawing. The percentage of sperm that underwent an AR when incubated with PVL post thawing in Lake (45.4±2.7) was lower (P<0.05) than that of fresh semen (55.3±3.1), whereas there were no differences (P>0.05) with BPSE. It is concluded that Lake diluent was more efficient for fresh seminal storage, while BPSE diluent was more efficient for seminal cryopreservation in Harris hawk.Keywords: acrosome, cryopreservation, sperm. RESUMEN.El almacenamiento seminal in vitro ha contribuido a la reproducción de aves silvestres en cautiverio. Sin embargo es necesario predecir su capacidad fertilizante. Los indicadores de reacción acrosomal (RA) espermática se han relacionado con su capacidad fertilizante. Por lo anterior, el objetivo de este trabajo fue evaluar la capacidad de reacción acrosomal in vitro en espermatozoides de halcón Harris, conservados en fresco y criopreservados. Se obtuvieron 64 eyaculados, los cuales fueron evaluados en fresco y posdescongelación, 32 se diluyeron con medio Beltsville Poultry Semen Extender (BPSE) y 32 utilizando medio Lake. Se evaluaron 64 eyaculados en fresco y posdescongelación; para la evaluación espermática básica se utilizó la tinción de eosina-nigrosina, y capacidad de RA fue mediante su inducción con membrana perivitelina (MPV). La movilidad espermática en semen fresco fue mayor (P<0,05) cuando se usó el medio BPSE (37,9±1,7) en comparación con el medio Lake (30,9±1,7). Sin embargo, la movilidad en ambos diluyentes disminuyó (P<0,05) posdescongelación. En semen fresco, el porcentaje de RA sin incubar con MPV fue mayor (P<0,05) con medio BPSE (14,1±1,7) al porcentaje con medio Lake (6,8±2,5). En semen descongelado los promedios de RA fueron similares (P>0,05). El porcentaje de espermatozoides con RA en el medio Lake, incubados con MPV posdescongelación (45,4±2,7), fue menor (P<0,05), comparado en fresco (55,3±3,1). Se concluye que el diluyente Lake fue más eficaz para la conservación en fr...

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Effects of calpain and Rho-kinase inhibitors on the acrosome reaction and motility of fowl spermatozoa in vitro

Cited by 13 publications

References 64 publications

A reappraisal of the factors involved in in vitro initiation of the acrosome reaction in chicken spermatozoa

A reappraisal of the factors involved in in vitro initiation of the acrosome reaction in chicken spermatozoa

Potential Involvement of Several Signaling Pathways in Initiation of the Chicken Acrosome Reaction1

Evaluation of two diluents for the storage of fresh and cryopreserved semen of Harris hawk (Parabuteo unicinctus)

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