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Heat shock transcription factor 1 (HSF1) is the main regulator of the stress response that triggers the transcription of several genes encoding heat shock proteins (Hsps). Hsps act as molecular chaperones involved in protein folding, stability, and trafficking. HSF1 is highly expressed in oocytes and Hsf1 knock-out in mice revealed that in the absence of stress this factor plays an important role in female reproduction. We previously reported that Hsf1-/- females produce oocytes but no viable embryos. Consequently, we asked whether oocytes require HSF1 to regulate a particular set of Hsps necessary for them to develop. We find that Hsp90α (Hspaa1) is the major HSF1-dependent chaperone inasmuch as Hsf1 knock-out resulted in Hsp90-depleted oocytes. These oocytes exhibited delayed germinal vesicle breakdown (or G2/M transition), partial meiosis I block, and defective asymmetrical division. To probe the role of Hsp90α in this meiotic syndrome, we analyzed meiotic maturation in wild-type oocytes treated with a specific inhibitor of Hsp90, 17-allylamino-17-demethoxy-geldanamycin, and observed similar defects. At the molecular level we showed that, together with these developmental anomalies, CDK1 and MAPK, key meiotic kinases, were significantly disturbed. Thus, our data demonstrate that HSF1 is a maternal transcription factor essential for normal progression of meiosis.

The effect of the sulfhydryl reagent, thimerosal (TMS) on meiosis resumption in germinal vesicle (GV)-stage denuded mouse oocytes was studied. It irreversibly inhibits both GV breakdown (GVBD) and the first polar body (pb1) extrusion in concentration- and time-dependent manners, the most striking result being the very early and narrow temporal window during which denuded primary oocytes released from their follicle are susceptible to a pulse of the drug. This inhibition is bypassed by dithiothreitol (DTT) with an efficiency declining with time, while thiosalicylic acid (TA), an analog of TMS devoid of the mercury atom, has no effect on meiosis reinitiation. These results strongly suggest that the inhibitory effect of TMS is a consequence of its sulfhydryl group oxidising activity. The molecular target(s) of this inhibitory oxidation should however be identified. In contrast to DTT, okadaic acid (OA), known to bypass the inhibitory effect of drugs interfering with protein kinase activities, only induces chromatin condensation and GVBD in TMS-pulsed oocytes with a delay of about 8 hr as compared to the control situation. This confirms that a very early thiol oxidation induced by TMS exerts a much more dramatic effect on resumption on meiosis than any pharmacological manipulation of protein kinase activities leading to activation of MPF.

A pulse of thimerosal (TMS), a sulfhydryl reagent, induces an instantaneous, complete and long-lasting microtubule interphasic network disassembly in mouse primary oocytes, correlated with the irreversible inhibition of meiosis reinitiation This inhibition is bypassed by dithiothreitol (DTT) while thiosalicylic acid, an analog of TMS, does induce neither microtubules depolymerisation nor inhibition of reinitiation and resumption of meiosis. This strongly suggests that the dramatic and pleiotropic inhibitory effect of TMS is specifically related to its sulfhydryl group oxidising activity of critical molecules among which tubulin. In contrast to DTT, okadaic acid (OA), known to bypass the inhibitory effect of drugs interfering with protein kinase activities, induces a late chromatin condensation and GVBD in TMS-pulsed oocytes as compared to the control situation, with no significant concomitant microtubule assembly. These cytological features are suggested to be indirectly induced by a late MAPK activation and confirm that a very early thiol oxidation induced by TMS exerts a much more dramatic effect on resumption of meiosis than any pharmacological manipulation of protein kinase activities leading to activation of MPF. Finally, taxol was shown to promote tubulin polymerisation even when microtubules were irreversibly disassembled by thiol oxidation but fails to restore the ability to undergo maturation.