Chiavegato A, Capriani A, Azzarello G, Vinante O, Pauletto P, Sartore S
Department of Biomedical Sciences, University of Padua, Padua, Italy.
Cell Tissue Res. 1996 Jan;283(1):7-18. doi: 10.1007/s004410050507.
The putatative effects of different estrogen levels on the expression of non-muscle myosin isoforms in rabbit myometrium have been investigated using three monoclonal anti-platelet myosin heavy chain (MyHC) antibodies (NM-F6, NM-G2, and NM-A9). Western blotting analysis of proteolytic digests of human platelet actomyosin indicates that these antibodies are specific for three distinct epitopes. Comparative immunofluorescence tests on cultered human fibroblasts with polyclonal sequence-specific anti-MyHCA antibody suggest that the patterns of NM-F6, NM-.G2 and NM-A9, although similar, do not overlap with that of type-A MyHC. Distribution of NM myosin isoforms has been studied in indirect immunofluorescence assays using cryosections of tissues from rabbits at various stages of development, pregnancy, or from ovariectomized, 17beta-estradiol-treated ovariectomized, and human chorionic gonadotropin-treated animals. Non-muscle myosin antigenicity is still present in the myometrium when the female becomes sexually competent. The immunoreactivity of non-muscle myosin for NM-F6 is steroid-independent, since it does not change with pregnancy or ovariectomy, but that of NM-G2 is estrogen-dependent; the latter disappears during pregnancy and in ovariectomized animals treated with estradiol, whereas it is expressed in ovariectomized rabbits. Although non-muscle myosin immunoreactivity for NM-A9 is detectable under all the experimental conditions, it can assume different patterns of intracellular distribution in vitro (punctate vs filamentous), depending on culture conditions and the presence of estrogens.
利用三种抗血小板肌球蛋白重链(MyHC)单克隆抗体(NM-F6、NM-G2和NM-A9),研究了不同雌激素水平对兔子宫肌层非肌肉肌球蛋白亚型表达的潜在影响。对人血小板肌动球蛋白的蛋白水解消化产物进行的蛋白质印迹分析表明,这些抗体对三个不同的表位具有特异性。用多克隆序列特异性抗MyHCA抗体对培养的人成纤维细胞进行的比较免疫荧光试验表明,NM-F6、NM-G2和NM-A9的模式虽然相似,但与A型MyHC的模式不重叠。使用处于不同发育阶段、妊娠阶段的兔子,或去卵巢、经17β-雌二醇处理的去卵巢兔子以及经人绒毛膜促性腺激素处理的动物的组织冰冻切片,通过间接免疫荧光试验研究了NM肌球蛋白亚型的分布。当雌性动物具备性能力时,子宫肌层中仍存在非肌肉肌球蛋白抗原性。非肌肉肌球蛋白对NM-F6的免疫反应性不依赖于类固醇,因为它不会随妊娠或去卵巢而改变,但NM-G2的免疫反应性依赖于雌激素;后者在妊娠期间以及经雌二醇处理的去卵巢动物中消失,而在去卵巢的兔子中表达。尽管在所有实验条件下都能检测到非肌肉肌球蛋白对NM-A9的免疫反应性,但根据培养条件和雌激素的存在情况,它在体外可呈现不同的细胞内分布模式(点状与丝状)。