Laboratorio de Medicina Reproductiva, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rosario, Argentina.
Cell Tissue Res. 2020 Jun;380(3):657-667. doi: 10.1007/s00441-020-03180-2. Epub 2020 Feb 28.
The oviduct is a dynamic organ that suffers changes during the oestrous cycle and modulates gamete and embryo physiology. We analyse the possible existence of Protein kinase A (PKA)-dependent hormone-regulated pathways in porcine ampulla and primary cell cultures by 2D-electrophoresis/Western blot using anti-phospho PKA substrate antibodies. Differential phosphorylation was observed for ten proteins that were identified by mass spectrometry. The results were validated for five of the proteins: Annexin A5, Calumenin, Glyoxalase I and II and Enolase I. Immunofluorescence analyses show that Calumenin, Glyoxalase II and Enolase I change their localisation in the oviductal epithelium through the oestrus cycle. The results demonstrate the existence of PKA hormone-regulated pathways in the ampulla epithelium during the oestrus cycle.
输卵管是一个动态器官,在发情周期中会发生变化,并调节配子和胚胎的生理机能。我们通过使用抗磷酸化 PKA 底物抗体的 2D 电泳/免疫印迹分析,来研究在猪输卵管和原代细胞培养物中是否存在蛋白激酶 A(PKA)依赖性激素调节途径。通过质谱法鉴定出十种差异磷酸化的蛋白质。对其中的五种蛋白质:膜联蛋白 A5、钙网蛋白、甘油醛-3-磷酸脱氢酶和烯醇酶 I 进行了验证。免疫荧光分析表明,钙网蛋白、甘油醛-3-磷酸脱氢酶和烯醇酶 I 的定位在发情周期中会发生变化。这些结果表明在发情周期中,PKA 激素调节途径存在于输卵管上皮中。