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PLoS One
2011 Jan 07;61:e16220. doi: 10.1371/journal.pone.0016220.
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Effects of thioglycolic acid on parthenogenetic activation of Xenopus oocytes.
Wang Z
,
Ren X
,
Wang D
,
Guan Y
,
Xia L
.
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Existing in Permanent-wave solutions (PWS), thioglycolic acid (TGA) is widely used in hairdressing industry for its contribution to hair styling. However, the toxicity of TGA, especially its reproductive toxicity, gradually calls the attention of more and more researchers. In this work, xenopus oocytes were pretreated with different concentration of TGA, and then activated by calcium ionophore A23187. During culture, the oocytes activation rates were taken note at different time after adding calcium ionophore A23187. At the end of the culture period, the nuclear status was detected under confocal microscope. In addition, some other samples were collected for Western-Blotting analysis. TGA significantly inhibited the oocytes activation rate and pronuclear formation. It may be resulted from the inhibition of the degradation of p-ERK1, Mos and CyclinB2. TGA inhibits in vitro parthenogenetic activation of xenopus oocytes with inhibited the degradation of proteins involved in mitogenic-activated protein kinase (MAPK) and maturation-promoting factor (MPF) pathways.
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21297954
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Figure 2. Effect of TGA on the fertilization coat formation of xenopus oocytes.
(A) An unfertilized egg which arrested in MII, there is a white spot on the animal pole; (B) An parthenogenetic activated egg induced by 2 /ml calcium ionophore A23187 without TGA treatment. (C) An partheogenetic activated egg induced by 2 /ml calcium ionophore A23187 pretreated with 125 /ml TGA for 2 h.
Figure 1. Effect of TGA on calcium ionophore A23187 induced parthenogenetic activation rate of xenopus oocytes in vitro.Xenopus MII oocytes were firstly pretreated with different dose of TGA (0, 5, 25, 125 µg/ml), after rinsed off TGA with MMR, the oocytes were then activated by 2 µg/ml calcium ionophore A23187. Oocytes activation was closely monitored after the addition of A23187. The activation rates were counted at the indicated time point (10, 20, 30, 60 min after A23187 addition).
Figure 3. Effect of TGA on the pronuclear formation of xenopus oocytes.Ooytes were activated with A23187, 1 h after the addition of the stimulus, oocytes were fixed, stained with Hochest 33258 and examined under confocal microscope. (A)Pronucleus could be observed (marked by the white arrow) in the control group which was not treated with TGA. (B) Oocytes treated with 125 µg/ml TGA failed to form pronucleus.
Figure 4. Expression of proteins involve in MPF and MAPK in xenopus oocytes at different time after the addition of calcium ionophore A23187 without TGA treatment.
Figure 5. Effects of TGA on the expression of proteins involved in MPF and MAPK during the process of parthenogenetic activation.Oocytes were homogenized 1 h after calcium ionophore A23187 addition and immunoblotted with the indicated antibodies. Actin was used as a control for protein loading.
Figure 6. Effects of TGA on the expression of proteins involved in MPF and MAPK during the process of parthenogenetic activation.The bar graph shows the relative quantification of band intensity. Significance indicated by *p<.05, **p<.01, compared with TGA 0 µg/ml group.
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