ABSTRACT
Calcineurin is required for oocyte exit from meiotic block in metaphase II (MII) stage in invertebrates and also in lower vertebrates. However, the role of calcineurin in mammalian oocyte activation is still unclear. The aim of this study was to determine whether calcineurin is involved in the processes regulating porcine oocyte activation. Indirect immunofluorescence demonstrated localization of both calcineurin subunits, CnA and CnB, especially in the cortex area of MII oocytes, in vitro fertilized and also parthenogenetically activated oocytes. After activation, the fluorescence intensity of the protein in the cortex area of oocytes remains unchanged; the protein calcineurin in the cytoplasm was recorded mainly around the pronuclei. Treatment of matured oocytes with calcineurin inhibitors, cyclosporin A (CsA) and hymenistatin I (HS-I), followed by activation with calcium ionophore A23187, significantly decreased the rate of activated oocytes compared to oocytes that were treated only with calcium ionophore (Ca-Io), (CsA+Ca-Io 25.0% v. Ca-Io 83.3%; HS-I+Ca-Io 32.5% v. Ca-Io 85.0%). Compared to the control, CsA treatment of matured oocytes followed by activation with Ca-Io did not affect the activity level of metaphase-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) in activated oocytes evaluated by kinase activity assay. Simultaneous staining of calcineurin and cortical granule content in matured oocytes showed that calcineurin distributed in the cortical area of the oocyte has not been colocalized with cortical granules content. On the other hand, the calcineurin inhibition before parthenogenetic activation leads to a reduction of the cortical reaction level compared to oocytes that were not treated with CsA (complete exocytosis: CsA+Ca-Io 2.6% v. Ca-Io 83.9%; sum of cortical granule brightness: CsA + Ca-Io 0.69 v. Ca-Io 0.15). Our results showed that calcineurin is involved in the process of pig oocyte activation and cortical granule exocytosis; however this regulation seems to be MPF and MAPK independent.
Subject(s)
Calcineurin/pharmacology , Oocytes/drug effects , Swine/physiology , Animals , Calcium Ionophores/pharmacology , Exocytosis , Fertilization in Vitro , In Vitro Oocyte Maturation Techniques/veterinary , Metaphase , Oocytes/metabolism , Parthenogenesis/physiologyABSTRACT
We tested the effect of two different concentrations (150µg/l and 0.15µg/l) of mycotoxin zearalenone (ZEA) on the reproductive parameters and expression of testicular genes in male mice. In adult males, no reduction of body or reproductive organ weight was observed, and the seminiferous tubules were morphologically normal with ongoing spermatogenesis. However, we found decreased sperm concentration, increase of morphologically abnormal spermatozoa and increased binding of apoptotic marker annexin V. This study was also focused on the evaluation of gene expression profiles of 28 genes playing important roles during the processes occurring in the testicular tissue. We detected changes in the expression of genes important for proper spermatogenesis. Surprisingly, we observed a stronger effect after exposure to the lower dose of ZEA.
Subject(s)
Estrogens, Non-Steroidal/toxicity , Testis/drug effects , Zearalenone/toxicity , Animals , Apoptosis/drug effects , Gene Expression/drug effects , Male , Mice , Spermatogenesis/drug effects , Spermatozoa/drug effects , Spermatozoa/pathology , Testis/metabolismABSTRACT
The authors assessed the levels of lactate and 2,3 diphosphoglycerate in the umbilical blood of 105 full-term neonates. A significant increase was found in the levels of 2,3 diphosphoglycerate in newborns of mothers with a history of imminent chronic intrauterine hypoxia. The lactate levels of these newborns were the same as in children of mothers without a history showing a risk of intrauterine hypoxia. The psychomotor development of newborns with increased levels of 2,3 diphosphoglycerate was significantly more altered than in those with normal levels in their first year of life.
