Tseng Yung-Che, Chen Ruo-Dong, Lee Jay-Ron, Liu Sian-Tai, Lee Shyh-Jye, Hwang Pung-Pung
Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, Taiwan 11529, ROC.
Am J Physiol Regul Integr Comp Physiol. 2009 Aug;297(2):R275-90. doi: 10.1152/ajpregu.00180.2009. Epub 2009 May 20.
Glucose, a carbohydrate metabolite, plays a major role in the energy supply for fish iono- and osmoregulation, and the way that glucose is transported in ionocytes is a critical process related to the functional operations of ionocytes. Eighteen members of glucose transporters (GLUTs, SLC2A) were cloned and identified from zebrafish. Previously, Na(+),K(+)-ATPase-rich (NaR), Na(+)-Cl(-) cotransporter-expressing (NCC), H(+)-ATPase-rich (HR), and glycogen-rich (GR) cells have been identified to be responsible for Ca(2+) uptake, Cl(-) uptake, Na(+) uptake, and the energy deposition, respectively, in zebrafish skin/gills. The purpose of the present study was to test the hypothesis of whether GLUT isoforms are specifically expressed and function in ionocytes to supply energy for ion regulatory mechanisms. On the basis of translational knockdown of foxi3a/3b (2 transcriptional factors related to the ionocytes' differentiation) and triple in situ hybridization/immunocytochemistry, 3 GLUT isoforms, zglut1a, -6, and -13.1, were specifically localized in NaR/NCC cells, GR cells, and HR cells, respectively. mRNA expression of zglut1a in embryos and adult gills were stimulated by the low Ca(2+) or low Cl(-) freshwater, which has been previously reported to upregulate the functions (monitored by epithelial Ca(2+) channel, NCC mRNA) of NaR/NCC cells, respectively while that of zglut13.1 was stimulated only by low Na(+), a situation to upregulate the function (monitored by carbonic anhydrase 15a mRNA) of HR cells. On the other hand, ambient ion compositions did not affect the zglut6 mRNA expression. Taken together, zGLUT1a, -6, and 13.1, the specific transporters in NaR/NCC cells, GR cells, and HR cells, may absorb glucose into the respective cells to fulfill different physiological demands.
葡萄糖作为一种碳水化合物代谢产物,在鱼类离子调节和渗透调节的能量供应中发挥着重要作用,而葡萄糖在离子细胞中的运输方式是与离子细胞功能运作相关的关键过程。从斑马鱼中克隆并鉴定出了18种葡萄糖转运蛋白(GLUTs,SLC2A)成员。此前,已确定富含Na(+)、K(+)-ATP酶(NaR)、表达Na(+)-Cl(-)共转运蛋白(NCC)、富含H(+)-ATP酶(HR)和富含糖原(GR)的细胞分别负责斑马鱼皮肤/鳃中的Ca(2+)摄取、Cl(-)摄取、Na(+)摄取和能量沉积。本研究的目的是验证GLUT异构体是否在离子细胞中特异性表达并发挥功能,为离子调节机制提供能量这一假设。基于foxi3a/3b(与离子细胞分化相关的2种转录因子)的翻译敲低以及三重原位杂交/免疫细胞化学技术,3种GLUT异构体,即zglut1a、-6和-13.1,分别特异性定位于NaR/NCC细胞、GR细胞和HR细胞中。低Ca(2+)或低Cl(-)的淡水可刺激胚胎和成体鳃中zglut1a的mRNA表达,此前已有报道称,低Ca(2+)或低Cl(-)淡水分别上调NaR/NCC细胞的功能(以上皮Ca(2+)通道、NCC mRNA监测),而zglut13.1的mRNA表达仅受低Na(+)刺激,低Na(+)可上调HR细胞的功能(以碳酸酐酶15a mRNA监测)。另一方面环境离子组成不影响zglut6的mRNA表达。综上所述,NaR/NCC细胞、GR细胞和HR细胞中的特异性转运蛋白zGLUT1a、-6和13.1可能将葡萄糖吸收到各自细胞中以满足不同的生理需求。