Center for Neurobiology of Stress, CURE/Digestive Diseases Research Center, USA.
Biochem Biophys Res Commun. 2010 Jan 29;392(1):67-71. doi: 10.1016/j.bbrc.2009.12.169. Epub 2010 Jan 7.
The enzyme that acylates ghrelin was recently identified in mice as the fourth member of the membrane-bound O-acyltransferases superfamily (MBOAT4) and named ghrelin-O-acyltransferase (GOAT). Only one report showed GOAT mRNA expression in ghrelin-expressing cells of the mouse stomach. We investigated the distribution of GOAT protein in peripheral tissues and co-expression with endocrine markers in the gastric mucosa using a custom-made anti-GOAT antibody. Tissues were collected from male Sprague-Dawley rats and C57BL/6 mice. Western blot revealed two immunoreactive bands in rat and mouse gastric corpus mucosal proteins, a 50 kDa band corresponding to the GOAT protein and a 100 kDa band likely corresponding to a dimer. Western blot also detected GOAT in the plasma and levels were strongly increased after 24-h fasting in mice and slightly in rats. GOAT-immunoreactive cells were located in the gastric corpus mucosa and the anterior pituitary gland, whereas other peripheral tissues of rats and mice examined were negative. In mice, GOAT-immunoreactive cells were mainly distributed throughout the middle portion of the oxyntic glands, whereas in rats they were localized mainly in the lower portion of the glands. Double labeling showed that 95+/-1% of GOAT-immunoreactive cells in mice co-labeled with ghrelin, whereas in rats only 56+/-4% of GOAT-positive cells showed co-expression of ghrelin. The remainder of the GOAT-immunopositive cells in rats co-expressed histidine decarboxylase (44+/-3%). No co-localization was observed with somatostatin in rats or mice. These data suggest species differences between rats and mice in gastric GOAT expression perhaps resulting in a different role of the MBOAT4 enzyme in the rat stomach. Detection of GOAT in the plasma raises the possibility that ghrelin octanoylation may occur in the circulation and the fasting-induced increase in GOAT may contribute to the increase of acylated ghrelin after fasting.
最近在小鼠中发现,酰化胃饥饿素的酶是膜结合 O-酰基转移酶超家族(MBOAT4)的第四种成员,并被命名为胃饥饿素-O-酰基转移酶(GOAT)。仅有一份报告显示 GOAT mRNA 在小鼠胃中表达胃饥饿素的细胞中表达。我们使用定制的抗 GOAT 抗体研究了 GOAT 蛋白在周围组织中的分布及其与胃黏膜内分泌标志物的共表达。组织取自雄性 Sprague-Dawley 大鼠和 C57BL/6 小鼠。Western blot 显示,大鼠和小鼠胃体黏膜蛋白中有两条免疫反应性条带,50 kDa 条带对应于 GOAT 蛋白,100 kDa 条带可能对应于二聚体。Western blot 还在血浆中检测到 GOAT,禁食 24 小时后,小鼠的水平明显升高,大鼠的水平略有升高。GOAT-免疫反应性细胞位于胃体黏膜和垂体前叶,而检查的大鼠和小鼠其他外周组织均为阴性。在小鼠中,GOAT-免疫反应性细胞主要分布在整个胃底腺的中部,而在大鼠中则主要分布在胃底腺的下部。双标记显示,95+/-1%的小鼠 GOAT-免疫反应性细胞与胃饥饿素共标记,而在大鼠中,只有 56+/-4%的 GOAT 阳性细胞显示与胃饥饿素共表达。大鼠中其余的 GOAT-免疫阳性细胞共表达组氨酸脱羧酶(44+/-3%)。在大鼠或小鼠中均未观察到与生长抑素的共定位。这些数据表明,胃中 GOAT 的表达在大鼠和小鼠之间存在种间差异,这可能导致 MBOAT4 酶在大鼠胃中的作用不同。在血浆中检测到 GOAT 提示,胃饥饿素的辛酰化可能发生在循环中,禁食诱导的 GOAT 增加可能导致禁食后酰化胃饥饿素增加。