Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways

Rodriguez, P.; O’Flaherty, Cristián; Beconi, M.T.; Beorlegui, N.

doi:10.1016/j.anireprosci.2004.05.018

Cited by 47 publications

(44 citation statements)

References 41 publications

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“…An association has been reported between sperm motility and the precursor of peroxynitrite, nitric oxide, high concentrations of which were associated with reduced sperm motility in in vitro studies (Tomlinson et al, 1992;Herrero et al, 1994;Rosselli et al, 1995;Weinberg et al, 1995;Rodriguez et al, 2005) and in asthenozoospermic patients (Aksoy et al, 2002;Balercia et al, 2004). Thus, if NO can generate ONOO 2 in vivo, then this highly reactive radical could ultimately be responsible for impaired sperm function.…”

Section: Discussionmentioning

confidence: 99%

Peroxynitrite-mediated nitrosative stress decreases motility and mitochondrial membrane potential in human spermatozoa

Uribe

Boguen

Treulén

et al. 2014

MHR: Basic Science of Reproductive Medicine

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Nitrosative stress is produced by high levels of reactive nitrogen species (RNS). The RNS include peroxynitrite, a highly reactive free radical produced from a diffusion-controlled reaction between nitric oxide and superoxide anion. Peroxynitrite causes nitration and oxidation of lipids, proteins and DNA, and is thus considered an important pathogenic mechanism in various diseases. Although high levels of peroxynitrite are associated with astenozoospermia, few reports exist regarding the in vitro effect of high levels of this RNS on human sperm. The aim of this study was to evaluate the in vitro effect of nitrosative stress caused by peroxynitrite on the viability, motility and mitochondrial membrane potential of human spermatozoa. To do this, human spermatozoa from healthy donors were exposed in vitro to 3-morpholinosydnonimine (SIN-1), a molecule that generates peroxynitrite. Incubations were done at 378C for up to 4 h with SIN-1 concentrations between 0.2 and 1.0 mmol/l. Generation of peroxynitrite was confirmed using dihydrorhodamine 123 (DHR) by spectrophotometry and flow cytometry. Sperm viability was assessed by propidium iodide staining; sperm motility was analyzed by CASA, and the state of mitochondrial membrane potential (DCm) by JC-1 staining. Viability and DCm were measured by flow cytometry. The results showed an increase in DHR oxidation, demonstrating the generation of peroxynitrite through SIN-1. Peroxynitrite decreased progressive and total motility, as well as some sperm kinetic parameters. Mitochondrial membrane potential also decreased. These alterations occurred with no decrease in sperm viability. In conclusion, peroxynitriteinduced nitrosative stress impairs vital functions in the male gamete, possibly contributing to male infertility.

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Section: Discussionmentioning

confidence: 99%

Peroxynitrite-mediated nitrosative stress decreases motility and mitochondrial membrane potential in human spermatozoa

Uribe

Boguen

Treulén

et al. 2014

MHR: Basic Science of Reproductive Medicine

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“…Decreased and controlled ROS help in sperm development and maturation (Kumar et al, 1991;Aitken and Vernet, 1998), capacitation (de Lamirande and Cagnon, 1993, 1995O'Flaherty et al, 2003O'Flaherty et al, , 2006 de , acrosome reaction (de Lamirande et al, 1998;Rodriguez et al, 2005), and sperm-zona interaction (Said et al, 2004). Epididymal maturation, capacitation, and acrosome reaction in spermatozoa are linked with progressive alterations in Superoxide production.…”

Section: Discussionmentioning

confidence: 95%

Modulation of nicotinamide adenine dinucleotide phosphate oxidase activity through sequential posttranslational modifications of p22 phagocytic oxidase during capacitation and acrosome reaction in goat spermatozoa1

Chandrasekhar

Laloraya

Kumar

2011

Journal of Animal Science

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Superoxide anion radical, produced in low quantities, plays a positive role in sperm function. Spermatozoa produce superoxide anion radical during posttesticular development, which shows an abrupt increase during capacitation. The NAD phosphate oxidase (NOX) family members NOX2 and NOX5 are the 2 enzymes implicated in superoxide production in spermatozoa. We examined the organization of NOX2 in goat spermatozoa during epididymal maturation, capacitation, and acrosome reaction. Spermatozoa from testis, caput epididymidis, corpus epididymidis, and cauda epididymidis possessed components of the phagocytic oxidase (PHOX; i.e., gp91phox, p22phox, p67phox, p47phox, p40phox), and ras-related C3 botulinum toxin substrate 1/2 (Rac1/2) on spermatozoa, and their concentrations did not show significant alterations during epididymal maturation. During capacitation in vitro, p22phox underwent Thr-phosphorylation, which resulted in a mobility shift of the corresponding band toward greater molecular mass. The Rac1/2 also showed a mobility shift from 32 to 23 kDa during capacitation. During progesterone-induced acrosome reaction, the spermatozoa experienced a total loss of p22phox and p47phox. The p47phox, but not p22phox, was detected in the exocytic vesicles of the acrosome. The Thr-phosphorylated form of p22phox was ubiquitinated and degraded through proteasome-mediated pathways in goat sperm cell lysates. Thus, Thr phosphorylation of p22phox acts as a regulatory switch in goat spermatozoa that transiently activates the NOX2 system during capacitation and subsequently directs it for degradation through the ubiiquitin-proteasomal pathway during progesterone-induced acrosome reaction.

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“…O controle consistiu de espermatozoides cultivados em meio de capacitação sem o inibidor da enzima iNOS. Para saber se este efeito seria impedido pela adição de NO, acrescentaram-se 500M de SNP ao meio (Rodriguez et al, 2004), juntamente à concentração de AG que apresentou o efeito deletério (0,1M+SNP).…”

Section: Methodsunclassified

“…Além disso, durante a capacitação induzida pela heparina, espermatozoides sintetizam NO (Leal et al, 2009). A participação do NO na capacitação de espermatozoides bovinos tem sido descrita na integridade da membrana, vigor e motilidade durante a capacitação (Paes de Carvalho et al, 2003;Rodriguez et al, 2004;Leal et al, 2009 A iNOS é seletivamente inibida pela aminoguanidina (AG), além de amidinas (Nakamura et al, 2002). Thornalley (2003) demonstrou que a AG é um potente inibidor da iNOS na concentração de 31M e um fraco inibidor da nNOS e eNOS nas concentrações de 170 e 330M, respectivamente, enquanto Nakamura et al (2002) observaram que 10 e 100mM de AG diminuíram a maturação de oócitos contidos em ovários de ratas com quatro a sete folículos pré-ovulatórios.…”

Section: Introductionunclassified

Efeito da inibição da óxido nítrico sintase induzível na capacitação in vitro de espermatozoides bovinos

Ferreira-Berbari¹,

Caldas-Bussiere²,

Carvalho³

et al. 2010

Arq. Bras. Med. Vet. Zootec.

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RESUMOAvaliaram-se o papel do óxido nítrico (NO) por meio da inibição da enzima óxido nítrico sintase induzível (iNOS), após a adição da aminoguanidina (AG), na motilidade, no vigor e na integridade da membrana plasmática nos tempos de 15, 60, 120, 180, 240 e 300min e a atividade mitocondrial e a capacitação de espermatozoides bovinos após 300min de cultivo. Adicionaram-se diferentes concentrações (0,001, 0,01 e 0,1M) de AG durante a capacitação induzida pela heparina e 500M de nitroprussiato de sódio (SNP, doador de NO) à concentração deletéria. A adição de 0,1M de AG diminuiu a motilidade e o vigor espermático e a integridade da membrana (P<0,05). A adição de SNP ao meio de cultivo com 0,1M de AG somente reverteu a integridade da membrana após 300min. A inibição da síntese de NO pela adição de AG não alterou a atividade mitocondrial. A percentagem de oócitos penetrados com espermatozoides tratados com 0,01 e 0,1M de AG diminuiu 20,3 e 100%, respectivamente, em relação aos não tratados (controle) (P<0,05), contudo houve aumento de 15% na percentagem de oócitos desnudados penetrados com espermatozoides capacitados em presença de 0,1M de AG. Conclui-se que a inibição da síntese de NO pela AG diminuiu a qualidade espermática durante a capacitação de espermatozoides bovinos in vitro, exceto a atividade mitocondrial. Somente a integridade da membrana foi revertida após adição de NO, sugerindo diferentes vias de ação do NO na qualidade espermática ao longo da capacitação in vitro de espermatozoides bovinos.Palavras-chave: bovino, óxido nítrico, qualidade do espermatozoide, capacitação in vitro, aminoguanidina 15, 60, 120, 180, 240, and 300min and on mitochondrial activity and capacitation after 300min, respectively. Different concentrations, 0.001, 0.01, and 0.1M of AG were added during the heparin induced capacitation and sodium nitroprusside (SNP, 01 and 0.1M of AG diminished, 20.3 and 100%, respectively, in ABSTRACT The role of nitric oxide (NO) was evaluated by inhibition of inducible nitric oxide synthase (iNOS), with aminoguanidine (AG) on motility, vigor, and plasmatic membrane integrity of bovine spermatozoa culture after

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Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways

Cited by 47 publications

References 41 publications

Peroxynitrite-mediated nitrosative stress decreases motility and mitochondrial membrane potential in human spermatozoa

Peroxynitrite-mediated nitrosative stress decreases motility and mitochondrial membrane potential in human spermatozoa

Modulation of nicotinamide adenine dinucleotide phosphate oxidase activity through sequential posttranslational modifications of p22 phagocytic oxidase during capacitation and acrosome reaction in goat spermatozoa1

Efeito da inibição da óxido nítrico sintase induzível na capacitação in vitro de espermatozoides bovinos

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