Vitale María Leiza, Garcia Christopher J, Akpovi Casimir D, Pelletier R-Marc
Département de pathologie et biologie cellulaire, Faculté de Médecine, Université de Montréal, Montreal, Québec, Canada.
PLoS One. 2017 Jul 31;12(7):e0182495. doi: 10.1371/journal.pone.0182495. eCollection 2017.
Folliculostellate cell gap junctions establish a network for the transmission of information within the anterior pituitary. Connexins make up gap junction channels. Changes in connexin (Cx) turnover modify gap junction-mediated intercellular communication. We have reported that cytokines and hormones influence Cx43 turnover and coupling in folliculostellate cells and in the folliculostellate cell line TtT/GF. In addition, the expression of different connexins alters intercellular communication and connexins may have functions besides cell coupling. Here we assessed the expression, turnover and subcellular localization of Cx46 and Cx50 in the anterior pituitary and TtT/GF cells. Then, we assessed the impact of various natural (lactation, annual reproductive cycle, bFGF) and pathological (autoimmune orchitis, diabetes/obesity) conditions associated with altered anterior pituitary hormone secretion on Cx46 and Cx50. Anterior pituitary Cx46 and Cx50 expression and subcellular distribution were cell-dependent. Cx46 was expressed by folliculostellate, TtT/GF and endocrine cells. In the cytoplasm, Cx46 was chiefly associated with lysosomes. Variously sized Cx46 molecules were recovered exclusively in the TtT/GF cell nuclear fraction. In the nucleus, Cx46 co-localized with Nopp-140, a nucleolar factor involved in rRNA processing. Neither cytoplasmic nor nuclear Cx46 and Cx43 co-localized. Cx50 localized to folliculostellate and TtT/GF cells, and to the walls of blood capillaries, not to endocrine cells. Cx50 was cytoplasmic and associated with the cell membrane, not nuclear. Cx50 did not co-localize with Cx46 but it co-localized in the cytoplasm and co-immunoprecipitated with Cx43. Cx46 and Cx50 responses to various physiological and pathological challenges were different, often opposite. Cx46 and Cx43 expression and phosphorylation profiles differed in the anterior pituitary, whereas Cx50 and Cx43 were similar. The data suggest that Cx46 participates to cellular growth and proliferation and that Cx50, together with Cx43, contributes to folliculostellate cell coupling.
滤泡星状细胞间隙连接在前脑垂体内建立了一个信息传递网络。连接蛋白构成间隙连接通道。连接蛋白(Cx)周转的变化会改变间隙连接介导的细胞间通讯。我们已经报道,细胞因子和激素会影响滤泡星状细胞以及滤泡星状细胞系TtT/GF中Cx43的周转和偶联。此外,不同连接蛋白的表达会改变细胞间通讯,并且连接蛋白可能除了细胞偶联之外还具有其他功能。在这里,我们评估了Cx46和Cx50在前脑垂体和TtT/GF细胞中的表达、周转和亚细胞定位。然后,我们评估了与前脑垂体激素分泌改变相关的各种自然(泌乳、年度生殖周期、碱性成纤维细胞生长因子)和病理(自身免疫性睾丸炎、糖尿病/肥胖症)状况对Cx46和Cx50的影响。前脑垂体Cx46和Cx50的表达及亚细胞分布具有细胞依赖性。滤泡星状细胞、TtT/GF细胞和内分泌细胞表达Cx46。在细胞质中,Cx46主要与溶酶体相关。仅在TtT/GF细胞核组分中检测到大小各异的Cx46分子。在细胞核中,Cx46与Nopp-140共定位,Nopp-140是一种参与核糖体RNA加工的核仁因子。细胞质和细胞核中的Cx46与Cx43均未共定位。Cx50定位于滤泡星状细胞和TtT/GF细胞以及毛细血管壁,而非内分泌细胞。Cx50位于细胞质中,与细胞膜相关,而非细胞核。Cx50未与Cx46共定位,但在细胞质中共定位且与Cx43进行了共免疫沉淀。Cx46和Cx50对各种生理和病理挑战的反应不同,通常相反。前脑垂体中Cx46和Cx43的表达及磷酸化谱不同,而Cx50和Cx43相似。数据表明,Cx46参与细胞生长和增殖,并且Cx50与Cx43一起有助于滤泡星状细胞偶联。