Department of Biochemistry and Molecular Biology, USC Norris Comprehensive Cancer Center, 1441 Eastlake Ave., Rm. 5308, Los Angeles, CA 90089-9176, USA.
FASEB J. 2013 Mar;27(3):955-64. doi: 10.1096/fj.12-213330. Epub 2012 Nov 24.
To investigate the role of GRP78 in adipogenesis and metabolic homeostasis, we knocked down GRP78 in mouse embryonic fibroblasts and 3T3-L1 preadipocytes induced to undergo differentiation into adipocytes. We also created an adipose Grp78-knockout mouse utilizing the aP2 (fatty acid binding protein 4) promoter-driven Cre-recombinase. Adipogenesis was monitored by molecular markers and histology. Tissues were analyzed by micro-CT and electron microscopy. Glucose homeostasis and cytokine analysis were performed. Our results indicate that GRP78 is essential for adipocyte differentiation in vitro. aP2-cre-mediated GRP78 deletion leads to lipoatrophy with ∼90% reduction in gonadal and subcutaneous white adipose tissue and brown adipose tissue, severe growth retardation, and bone defects. Despite severe abnormality in adipose mass and function, adipose Grp78-knockout mice showed normal plasma triglyceride levels, and plasma glucose and insulin levels were reduced by 40-60% compared to wild-type mice, suggesting enhanced insulin sensitivity. The endoplasmic reticulum is grossly expanded in the residual mutant white adipose tissue. Thus, these studies establish that GRP78 is required for adipocyte differentiation, glucose homeostasis, and balanced secretion of adipokines. Unexpectedly, the phenotypes and metabolic parameters of the mutant mice, which showed early postnatal mortality, are uniquely distinct from previously characterized lipodystrophic mouse models.
为了研究 GRP78 在脂肪生成和代谢稳态中的作用,我们在诱导分化为脂肪细胞的小鼠胚胎成纤维细胞和 3T3-L1 前脂肪细胞中敲低了 GRP78。我们还利用 aP2(脂肪酸结合蛋白 4)启动子驱动的 Cre 重组酶创建了脂肪组织特异性 Grp78 敲除小鼠。脂肪生成通过分子标志物和组织学进行监测。通过 micro-CT 和电子显微镜分析组织。进行葡萄糖稳态和细胞因子分析。我们的结果表明,GRP78 是体外脂肪细胞分化所必需的。aP2-cre 介导的 GRP78 缺失导致脂肪萎缩,生殖腺和皮下白色脂肪组织和棕色脂肪组织减少约 90%,严重生长迟缓,骨骼缺陷。尽管脂肪质量和功能严重异常,但脂肪组织特异性 Grp78 敲除小鼠的血浆甘油三酯水平正常,与野生型小鼠相比,血浆葡萄糖和胰岛素水平降低了 40-60%,表明胰岛素敏感性增强。内质网在残留的突变白色脂肪组织中明显扩张。因此,这些研究表明 GRP78 是脂肪细胞分化、葡萄糖稳态和平衡的脂肪因子分泌所必需的。出乎意料的是,表现出早期出生后死亡率的突变小鼠的表型和代谢参数与以前表征的脂肪营养不良小鼠模型独特不同。