小鼠前脑线粒体蛋白的增龄性变化
Expression changes of mitochondrial protein with age in mouse forebrain development
目的 探讨小鼠前脑不同发育阶段线粒体蛋白的表达变化。 方法 分别取妊娠12.5 d（E12.5）及17.5 d（E17.5）的胎鼠、出生2 d的新生鼠、出生3周的幼鼠和2月龄成年鼠的前脑组织，利用QRT-PCR和Western blotting检测小鼠前脑发育不同阶段中COX IV、HSP60、OGDH、PDHE1α的mRNA和蛋白表达水平。 结果 在E12.5 ~2 d的前脑组织中COX IV mRNA呈低水平表达，3周时表达量明显升高并达到顶峰；HSP60和PDHE1α mRNA在E17.5表达明显增高，OGDH mRNA则在2 d显著上升，三者表达水平均于2月达到顶峰。COX IV、HSP60、OGDH的蛋白水平从E12.5 ~3周无明显差异，但在2月时表达显著上升；PDHE1α蛋白随前脑发育其表达水平不断提高，并于2月达到高峰。此外，COX IV、HSP60、OGDH、PDHE1α mRNA与蛋白表达水平呈正相关。 结论 COX IV、HSP60、OGDH和PDHE1α可能参与了小鼠前脑发育过程且与年龄密切相关。
Objective To investigate the changes of mitochondrial protein expression in different stages of mouse forebrain development. Methods The forebrain tissues of fetal mice with 12.5 and 17.5 days pregnancy (E12.5, E17.5), 2-day-old neonatal mice , 3-week-old young mice and 2-month-old mice were collected to detect the expression levels of COX IV, HSP60, OGDH, PDHE1α mRNA and protein in different development stages by QRT-PCR and Western blotting. Results The mRNA level of COX IV was low in forebrain tissue from E12.5 to 2 days, and it increased significantly and reached the peak at 3 weeks. The mRNA levels of HSP60 and PDHE1α were significantly increased at E17.5, and the OGDH mRNA level had an obvious rise at 2 days, and all three reached the peak at 2 months. The proteins of COX IV, HSP60 and OGDH showed no significant difference from E12.5 to 3 weeks, but increased evidently at 2 months; and the protein of PDHE1α increased significantly with forebrain development and peaked at 2 months. In addition, there was a positive correlation between mRNA and protein expression levels of COX IV, HSP60, OGDH, PDHE1α. Conclusion COX IV, HSP60, OGDH and PDHE1α may be involved in mouse forebrain development and be closely related to age.
[1] Sullivan EM, Pennington ER, Green WD, et al. Mechanisms by which dietary fatty acids regulate mitochondrial structure-function in health and disease[J]. Adv nutr, 2018, 9(3):247-262.
[2] Keating DJ. Mitochondrial dysfunction, oxidative stress, regulation of exocytosis and their relevance to neurodegenerative diseases[J]. J Neurochem, 2008, 104(2):298-305.
[3] Yang X, Wu J, Jing S, et al. Mitochondrial protein sulfenation during aging in the rat brain[J]. Biophys Rep, 2018, 4(2):104-113.
[4] Raposo N, Planton M, Péran P, et al. Florbetapir imaging in cerebral amyloid angiopathy-related hemorrhages[J]. Neurology, 2017, 89(7): 697-704.
[5] Mian AZ, Edasery D, Sakai O, et al. Radiological imaging features of the basal ganglia that may predict progression to hemicraniectomy in large territory middle cerebral artery infarct[J]. Neuroradiology, 2017, 59(5): 477-484.
[6] Thomson CE, McCulloch M, Sorenson A, et al. Myelinated, synapsing cultures of murine spinal cord-validation as an in vitro model of the central nervous system[J]. Eur J Neurosci, 2008, 28(8): 1518-1535.
[7] Alleyne T, Ignacio DN, Sampson VB, et al. Simulating the slow to fast switch in cytochrome c oxidase catalysis by introducing a loop flip near the enzyme's cytochrome c (substrate) binding site[J]. Biotechnol Appl Biochem, 2017, 64(5): 677-685.
[8] Madeira VM. Overview of mitochondrial bioenergetics[J]. Methods Mol Biol, 2012, 810:1-6.
[9] Wilson DF, Vinogradov SA. Mitochondrial cytochrome c oxidase: mechanism of action and role in regulating oxidative phosphorylation[J]. J Appl Physiol, 2014, 117(12): 1431-1439.
[10] 亢君君, 梁卫华, 黄晓峰, 等. 组织化学染色与免疫电镜双标记方法在检测大鼠脑干前包钦格复合体细胞色素氧化酶活性中的应用[J]. 细胞与分子免疫学杂志, 2017, 33(9): 1177-1181.
[11] Li Y, Park JS, Deng JH, et al. Cytochrome c oxidase subunit IV is essential for assembly and respiratory function of the enzyme complex[J]. J Bioenerg Biomembr, 2006, 38(5-6):283-291.
[12] Wy?ewski Z, Gregorczyk KP, Szczepanowska J, et al. Functional role of Hsp60 as a positive regulator of human viral infection progression[J]. Acta Virol, 2018, 62(1): 33-40.
[13]Hartl FU, Bracher A, Hayer-Hartl M. Molecular chaperones in protein folding and proteostasis[J]. Nature, 2011, 475(7356): 324-332.
[14]Matheoud D, Sugiura A, Bellemare-Pelletier A, et al. Parkinson's disease-related proteins PINK1 and parkin repress mitochondrial antigen presentation[J]. Cell, 2016, 166(2): 314-327.
[15]Papa L, Germain D. SirT3 regulates the mitochondrial unfolded protein response[J]. Mol Cellular Biol, 2014, 34(4): 699-710.
四川省教育厅自然科学重点项目(18ZA0516)
京ICP备08002354号-3/
| 〈 |
|
〉 |
