Tucker Nathan R, Ustyugov Alexey, Bryantsev Anton L, Konkel Michael E, Shelden Eric A
School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA.
Cell Stress Chaperones. 2009 Sep;14(5):521-33. doi: 10.1007/s12192-009-0105-1. Epub 2009 Feb 24.
Constitutive expression of Hsp27 has been demonstrated in vertebrate embryos, especially in developing skeletal and cardiac muscle. Results of several previous studies have indicated that Hsp27 could play a role in the development of these tissues. For example, inhibition of Hsp27 expression has been reported to cause defective development of mammalian myoblasts in vitro and frog embryos in vivo. In contrast, transgenic mice lacking Hsp27 develop normally. Here, we examined the distribution of Hsp27 protein in developing and adult zebrafish and effects of suppressing Hsp27 expression using phosphorodiamidate morpholino oligonucleotides (PMO) on zebrafish development. Consistent with our previous analysis of hsp27 messenger RNA expression, we detected the protein Hsp27 in cardiac, smooth, and skeletal muscle of both embryonic and adult zebrafish. However, embryos lacking detectable Hsp27 after injection of antisense hsp27 PMO exhibited comparable heart beat rates to that of control embryos and cardiac morphology was indistinguishable in the presence or absence of Hsp27. Loss of Hsp27 also had no effect on the structure of the skeletal muscle myotomes in the developing embryo. Finally, embryos injected with antisense hsp27 and scrambled control PMO displayed equal motility. We conclude that Hsp27 is dispensable for zebrafish morphogenesis but could play a role in long-term maintenance of heart and muscle tissues.
热休克蛋白27（Hsp27）的组成型表达已在脊椎动物胚胎中得到证实，尤其是在发育中的骨骼肌和心肌中。先前的几项研究结果表明，Hsp27可能在这些组织的发育中发挥作用。例如，据报道，抑制Hsp27表达会导致体外培养的哺乳动物成肌细胞和体内青蛙胚胎发育缺陷。相比之下，缺乏Hsp27的转基因小鼠发育正常。在此，我们研究了Hsp27蛋白在发育中的和成年斑马鱼中的分布，以及使用磷二酰胺吗啉代寡核苷酸（PMO）抑制Hsp27表达对斑马鱼发育的影响。与我们之前对hsp27信使核糖核酸表达的分析一致，我们在胚胎期和成年期斑马鱼的心脏、平滑肌和骨骼肌中检测到了Hsp27蛋白。然而，注射反义hsp27 PMO后缺乏可检测到的Hsp27的胚胎，其心跳速率与对照胚胎相当，并且在有或没有Hsp27的情况下，心脏形态没有区别。Hsp27的缺失对发育中胚胎的骨骼肌肌节结构也没有影响。最后，注射反义hsp27和乱序对照PMO的胚胎表现出相同的运动能力。我们得出结论，Hsp27对于斑马鱼的形态发生并非必需，但可能在心脏和肌肉组织的长期维持中发挥作用。
