Orihuela P A, Ríos M, Croxatto H B
Unidad de Reproducción y Desarrollo, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
Biol Reprod. 2001 Oct;65(4):1232-7. doi: 10.1095/biolreprod65.4.1232.
Previously, we found that the dose of estradiol (E2) required to accelerate egg transport increases 5- to 10-fold, in mated compared to cyclic rats. Here we examined protein synthesis in the oviduct of mated and cyclic rats following a single injection of E2 known to accelerate oviductal egg transport or after concomitant treatment with progesterone (P4) known to block this acceleration. On Day 1 of the cycle or pregnancy, E2, P4, or E2 + P4 were injected s.c., and 4 h later oviducts were removed and incubated for 8 h in medium with 35S-methionine. Tissue proteins were separated by SDS-PAGE, and protein bands were quantitated by fluorography and densitometry. In mated rats, E2 and P4 increased different protein bands and P4 did not affect the fluorographic pattern induced by E2. In contrast with mated rats, none of these treatments changed the fluorographic pattern of the oviductal proteins in cyclic rats. Estradiol-induced egg transport acceleration was then compared under conditions in which oviductal protein synthesis was suppressed. Mated and cyclic rats treated with equipotent doses of E2 for accelerating egg transport also received actinomycin D (Act D) locally. Estradiol-induced oviductal egg loss was partially blocked by Act D in mated but had no effect in cyclic rats. We conclude that the oviduct of mated and cyclic rats differs in that only the former responds with increased protein synthesis to a pulse of exogenous E2 and P4 and requires an intact protein synthesis machinery in order to accelerate egg transport in response to E2.
此前,我们发现,与处于发情周期的大鼠相比,交配后的大鼠加速卵子运输所需的雌二醇(E2)剂量增加了5至10倍。在此,我们检测了单次注射已知可加速输卵管卵子运输的E2后,以及同时用已知可阻断这种加速作用的孕酮(P4)处理后,交配后和处于发情周期的大鼠输卵管中的蛋白质合成情况。在发情周期或妊娠的第1天,皮下注射E2、P4或E2 + P4,4小时后取出输卵管,并在含有35S-甲硫氨酸的培养基中孵育8小时。通过SDS-PAGE分离组织蛋白,并用荧光自显影和光密度测定法定量蛋白条带。在交配后的大鼠中,E2和P4增加了不同的蛋白条带,且P4不影响E2诱导的荧光自显影模式。与交配后的大鼠相反,这些处理均未改变发情周期大鼠输卵管蛋白的荧光自显影模式。然后,在抑制输卵管蛋白合成的条件下,比较了E2诱导的卵子运输加速情况。用等效剂量的E2处理以加速卵子运输的交配后和处于发情周期的大鼠也局部接受了放线菌素D(Act D)。Act D部分阻断了交配后大鼠中E2诱导的输卵管卵子丢失,但对发情周期大鼠没有影响。我们得出结论,交配后和处于发情周期的大鼠的输卵管存在差异,仅前者对外源性E2和P4脉冲的反应是蛋白质合成增加,并且需要完整的蛋白质合成机制才能响应E2加速卵子运输。