Nitric oxide synthase isoforms and the effect of their inhibition on meiotic maturation of porcine oocytes

In this paper we assessed: (i) the change in nitric oxide synthase (NOS) isoforms' expression and intracellular localization and in NOS mRNA in porcine oocytes during meiotic maturation; (ii) the effect of NOS inhibition by Nω-nitro-l-arginine methyl ester (l-NAME) and aminoguanidine (AG) on me...

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Veröffentlicht in:Zygote (Cambridge) 2010-08, Vol.18 (3), p.235-244
Hauptverfasser: Chmelíková, Eva, Ješeta, Michal, Sedmíková, Markéta, Petr, Jaroslav, Tůmová, Lenka, Kott, Tomáš, Lipovová, Petra, Jílek, František
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container_title Zygote (Cambridge)
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creator Chmelíková, Eva
Ješeta, Michal
Sedmíková, Markéta
Petr, Jaroslav
Tůmová, Lenka
Kott, Tomáš
Lipovová, Petra
Jílek, František
description In this paper we assessed: (i) the change in nitric oxide synthase (NOS) isoforms' expression and intracellular localization and in NOS mRNA in porcine oocytes during meiotic maturation; (ii) the effect of NOS inhibition by Nω-nitro-l-arginine methyl ester (l-NAME) and aminoguanidine (AG) on meiotic maturation of cumulus–oocyte complexes (COC) as well as denuded oocytes (DO); and (iii) nitric oxide (NO) formation in COC. All three NOS isoforms (eNOS, iNOS and nNOS) and NOS mRNA (eNOS mRNA, iNOS mRNA and nNOS mRNA) were found in both porcine oocytes and their cumulus cells except for nNOS mRNA, which was not detected in the cumulus cells. NOS isoforms differed in their intracellular localization in the oocyte: while iNOS protein was dispersed in the oocyte cytoplasm, nNOS was localized in the oocyte cytoplasm and in germinal vesicles (GV) and eNOS was present in dots in the cytoplasm, GV and was associated with meiotic spindles. l-NAME inhibitor significantly suppressed metaphase (M)I to MII transition (5.0 mM experimental group: 34.9% MI, control group: 9.5% MI) and at the highest concentration (10.0 mM) also affected GV breakdown (GVBD); in contrast also AG inhibited primarily GVBD. The majority of the oocytes (10.0 mM experimental group: 60.8%, control group: 1.2%) was not able to resume meiosis. AG significantly inhibited GVBD in DO, but l-NAME had no significant effect on the GVBD of these cells. During meiotic maturation, NO is formed in COC and the NO formed by cumulus cells is necessary for the process of GVBD.
doi_str_mv 10.1017/S0967199409990268
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(ii) the effect of NOS inhibition by Nω-nitro-l-arginine methyl ester (l-NAME) and aminoguanidine (AG) on meiotic maturation of cumulus–oocyte complexes (COC) as well as denuded oocytes (DO); and (iii) nitric oxide (NO) formation in COC. 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During meiotic maturation, NO is formed in COC and the NO formed by cumulus cells is necessary for the process of GVBD.</abstract><cop>Cambridge, UK</cop><pub>Cambridge University Press</pub><pmid>20109267</pmid><doi>10.1017/S0967199409990268</doi><tpages>10</tpages></addata></record>
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source MEDLINE; Cambridge University Press Journals Complete
subjects Animals
Cumulus cells
Cumulus Cells - enzymology
Female
Meiosis
Meiotic maturation
Microscopy, Confocal
NG-Nitroarginine Methyl Ester - pharmacology
Nitric Oxide Synthase - antagonists & inhibitors
Nitric Oxide Synthase - genetics
Nitric Oxide Synthase - metabolism
NOS
Oocyte
Oocytes - drug effects
Oocytes - enzymology
Pig
Protein Isoforms - antagonists & inhibitors
Protein Isoforms - genetics
Protein Isoforms - metabolism
RNA, Messenger - metabolism
title Nitric oxide synthase isoforms and the effect of their inhibition on meiotic maturation of porcine oocytes
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