Cheng Henrique, Ellis Jayne, Kleinow Kevin M
Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA.
Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Comp Biochem Physiol C Toxicol Pharmacol. 2015 Dec;178:33-44. doi: 10.1016/j.cbpc.2015.09.010. Epub 2015 Sep 30.
Calcium signaling, from localized spikes to coordinated waves, are linked to cleavage, patterning, differentiation, and growth during embryonic development. The basis for control of these Ca(2+) signals is poorly defined. In this study, the expression and functionality of the transient receptor potential melastatin 4 protein (TRPM4), an ion channel that controls Ca(2+) entry into cells, was examined in the zebrafish embryo and adult. Originating with the human TRPM4 gene, Ensembl ortholog, NCBI BLAST, and Homologene searches identified a zebrafish TRPM4 "like" gene encoding a predicted protein of 1199 amino acids and sharing a 42-43% sequence identity with the mouse, rat, and human. Custom-designed zebrafish primers identified TRPM4 transcripts throughout the 0-123h period of embryonic development with greatest and lowest relative expression at 12 and 123h post-fertilization, respectively. Perforated patch clamp recordings in 27h embryonic cells revealed Ca(2+)-activated currents with the characteristics of those described for mammalian TRPM4. Similarly, TRPM4-like expression and functionality was observed in brain and liver cells from adult fish. These findings suggest that a TRPM4-like channel is available for Ca(2+) regulation during early development of the zebrafish.
钙信号传导，从局部峰值到协调波，与胚胎发育过程中的卵裂、模式形成、分化和生长相关。对这些Ca(2+)信号进行控制的基础仍不清楚。在本研究中，我们在斑马鱼胚胎和成年鱼中检测了瞬时受体电位褪黑素4蛋白（TRPM4）的表达和功能，TRPM4是一种控制Ca(2+)进入细胞的离子通道。通过对人类TRPM4基因、Ensembl直系同源基因、NCBI BLAST和同源基因数据库进行搜索，发现了一个斑马鱼TRPM4“类似”基因，该基因编码一个预测由1199个氨基酸组成的蛋白质，与小鼠、大鼠和人类的序列同一性为42-43%。定制设计的斑马鱼引物在胚胎发育的0-123小时内均检测到TRPM4转录本，在受精后12小时和123小时分别具有最高和最低的相对表达。对27小时胚胎细胞进行的穿孔膜片钳记录显示出具有哺乳动物TRPM4所描述特征的Ca(2+)激活电流。同样，在成年鱼的脑和肝细胞中也观察到了类似TRPM4的表达和功能。这些发现表明，在斑马鱼早期发育过程中，一种类似TRPM4的通道可用于Ca(2+)调节。
