Construction of mutant NDUFS7 plasmid and its effects on neural cells

SHEN Jie; ZHA Qian; GAO Xiangdong; CHEN Song

doi:10.11665/j.issn.1000-5048.20200512

Journal of China Pharmaceutical University > 2020 > 51(5): 599-606. > DOI: 10.11665/j.issn.1000-5048.20200512

SHEN Jie, ZHA Qian, GAO Xiangdong, CHEN Song. Construction of mutant NDUFS7 plasmid and its effects on neural cells[J]. Journal of China Pharmaceutical University, 2020, 51(5): 599-606. DOI: 10.11665/j.issn.1000-5048.20200512

Citation:

PDF (7980 KB)

Construction of mutant NDUFS7 plasmid and its effects on neural cells

Funds: This study was supported by the National Natural Science Foundation of China (No.81673435, No.81872850)

More Information

Received Date: April 03, 2020
Revised Date: June 28, 2020

Graphical Abstract

Abstract

Abstract

To investigate the effects of NDUFS7 gene mutation on neurons, the mutant plasmid of pcDNA3.1(+)-NDUFS7 Q208STOP was constructed and transfected into differentiated SH-SY5Y cells. The effect of transfecting mutant plasmid on the viability of dopaminergic neural cells was detected by MTT assay. The effect of transfection of mutant plasmid on apoptosis was detected by Annexin Ⅴ-FITC/PI staining followed by flow cytometry assay. The changes in the expression levels of apoptosis-related proteins Bax and Bcl-2 in cells after transfection of mutant plasmid were detected by Western blot. The effects of transfection of mutant plasmid on the mitochondrial membrane potential in differentiated SH-SY5Y cells and the intervention effect of antioxidant Trolox were examined using JC-1 fluorescent probe. The intervention effect of Trolox on the apoptosis of differentiated SH-SY5Y cells transfected with mutant plasmid was detected by PI/Hoechst staining. The results showed that the subunit mutation of mitochondrial complex I in dopaminergic neurons could lead to decreased neuronal viability and increased apoptosis, while antioxidants could alleviate the abnormal mitochondrial membrane potential and apoptosis caused by transfection of mutant plasmids, suggesting that transfection of mutant plasmid of NDUFS7 gene could lead to apoptosis by causing abnormal mitochondrial function in dopaminergic neurons.
- NDUFS7,
- dopaminergic neurons,
- cell apoptosis,
- mitochondrial membrane potential

FullText(HTML)

References (23)

References

[1]	. Cell Metab，2015，22（2）：204?206.
[2]	Roger AJ，Mu?oz-Gómez SA，Kamikawa R. The origin and diversification of mitochondria［J］. Curr Biol，2017，27（21）：R1177?R1192.
[3]	Walker JE. The NADH：ubiquinone oxidoreductase （complex I） of respiratory chains［J］. Q Rev Biophys，1992，25（3）：253?324.
[4]	Wirth C，Brandt U，Hunte C，et al. Structure and function of mitochondrial complex I［J］. Biochim Et Biophys Acta BBA-Bioenerg，2016，1857（7）：902?914.
[5]	Smeitink J，Sengers R，Trijbels F，et al. Human NADH：ubiquinone oxidoreductase［J］. J Bioenerg Biomembr，2001，33（3）：259?266.
[6]	Brandt U. Energy converting NADH：quinone oxidoreductase （complex I）［J］. Annu Rev Biochem，2006，75：69?92.
[7]	McBride HM，Neuspiel M，Wasiak S. Mitochondria：more than just a powerhouse［J］. Curr Biol，2006，16（14）：R551?R560.
[8]	Dudkina NV，Heinemeyer J，Sunderhaus S，et al. Respiratory chain supercomplexes in the plant mitochondrial membrane［J］. Trends Plant Sci，2006，11（5）：232?240.
[9]	Triepels RH，van den Heuvel LP，Loeffen JL，et al. Leigh syndrome associated with a mutation in the NDUFS7 （PSST） nuclear encoded subunit of complex I［J］. Ann Neurol，1999，45（6）：787?790.
[10]	Ma L，Zhang W，Tang JS，et al. No association between genetic polymorphisms of the NDUFS7 gene and schizophrenia in Han Chinese［J］. Psychiatr Genet，2013，23（1）：29?32.
[11]	Fiedorczuk K，Sazanov LA. Mammalian mitochondrial complex I structure and disease-causing mutations［J］. Trends Cell Biol，2018，28（10）：835?867.
[12]	Nonnekes J，Post B，Tetrud JW，et al. MPTP-induced Parkinsonism：an historical case series［J］. Lancet Neurol，2018，17（4）：300?301.
[13]	Langston JW. The MPTP story［J］. J Parkinsons Dis，2017，7（s1）：S11?S19.
[14]	Blaza JN，Vinothkumar KR，Hirst J. Structure of the deactive state of mammalian respiratory complex I［J］. Structure，2018，26（2）：312?319.
[15]	Chen S，Luo S，Zhang Z，et al. VHL-1 inactivation and mitochondrial antioxidants rescue C. elegans dopaminergic neurodegeneration［J］. Protein Cell，2019，10（8）：610?614.
[16]	Schultz W. Multiple dopamine functions at different time courses［J］. Annu Rev Neurosci，2007，30：259?288.
[17]	Bayer HM，Glimcher PW. Midbrain dopamine neurons encode a quantitative reward prediction error signal［J］. Neuron，2005，47（1）：129?141.
[18]	Floresco SB，West AR，Ash B，et al. Afferent modulation of dopamine neuron firing differentially regulates tonic and phasic dopamine transmission［J］. Nat Neurosci，2003，6（9）：968?973.
[19]	Lautrup S，Sinclair DA，Mattson MP，et al. NAD⁺ in brain aging and neurodegenerative disorders［J］. Cell Metab，2019，30（4）：630?655.
[20]	Surmeier DJ，Obeso JA，Halliday GM. Selective neuronal vulnerability in Parkinson disease［J］. Nat Rev Neurosci，2017，18（2）：101?113.
[21]	Ruan LH，Wang YH，Zhang X，et al. Mitochondria-associated proteostasis［J］. Annu Rev Biophys，2020，49：41?67.
[22]	Guo RY，Zong S，Wu M，et al. Architecture of human mitochondrial respiratory megacomplex I₂III₂IV₂［J］. Cell，2017，170（6）：1247-1257.
[23]	Lake NJ，Compton AG，Rahman S，et al. Leigh syndrome：one disorder，more than 75 monogenic causes［J］. Ann Neurol，2016，79（2）：190?203.

[1]	QIAN Qianqian, LI Guozhi, TIAN Hong, GAO Xiangdong. Design, preparation, and antitumor activity of fusion protein vaccine based on tumor antigen PBK[J]. Journal of China Pharmaceutical University, 2024, 55(5): 657-665. DOI: 10.11665/j.issn.1000-5048.2024030801
[2]	ZHOU Hongyou, GAO Xiangdong, YAO Wenbing, TIAN Hong. Screening of adjuvant for PD-L1 vaccine based on nitrated T cell epitope[J]. Journal of China Pharmaceutical University, 2024, 55(3): 397-403. DOI: 10.11665/j.issn.1000-5048.2023032801
[3]	LI Mengyuan, JIANG Xiaomeng, SUN Qinyi, GUO Wei. Research progress of CAR-T immunotherapy in solid tumors combined with new strategies[J]. Journal of China Pharmaceutical University, 2023, 54(4): 443-449. DOI: 10.11665/j.issn.1000-5048.2023030101
[4]	SHAO Shishuai, DUAN Shukang, TIAN Hong, YAO Wenbing, GAO Xiangdong. Design and antitumor activity of programmed cell death ligand 1 epitope peptide vaccine[J]. Journal of China Pharmaceutical University, 2023, 54(2): 245-254. DOI: 10.11665/j.issn.1000-5048.2023022803
[5]	WANG Zhenghao, GAO Yafeng, ZHANG Lianjun, LIU Chang. Research progress of T cell anti-tumor function regulated by endoplasmic reticulum stress[J]. Journal of China Pharmaceutical University, 2022, 53(5): 518-524. DOI: 10.11665/j.issn.1000-5048.20220502
[6]	YAO Zheng, LI Zihan, GAO Liming, HU Xing, CHEN Yan, PAN Wenqi, LI Qian. Advances of research on CAR-T cell immunotherapy for solid tumors[J]. Journal of China Pharmaceutical University, 2021, 52(4): 496-504. DOI: 10.11665/j.issn.1000-5048.20210413
[7]	XIA Xuefei, ZHANG Li, LUO Jianhua, YAO Wenbing, GAO Xiangdong, TIAN Hong. Design and antitumor activity of immune checkpoint B7-H3 epitope vaccine[J]. Journal of China Pharmaceutical University, 2021, 52(4): 472-479. DOI: 10.11665/j.issn.1000-5048.20210410
[8]	CHEN Hongmei, KANG Yanliang, LIU Li, YAO Wenbing, TIAN Hong. Effects of different immunogenic amino acids in PD-L1 vaccine on the differentiation of T cell subsets[J]. Journal of China Pharmaceutical University, 2020, 51(3): 349-356. DOI: 10.11665/j.issn.1000-5048.20200313
[9]	JIANG Liangliang, JIANG Tao, LUO Jianhua, YAO Wenbing, TIAN Hong. A novel human immune system mice model for assessing the immunogenicity of cancer vaccines[J]. Journal of China Pharmaceutical University, 2019, 50(6): 734-742. DOI: 10.11665/j.issn.1000-5048.20190615
[10]	SUN Zhan-yi, CAI Hui, HUANG Zhi-hua, SHI Lei, CHEN Yong-xiang, LI Yan-mei. Advances of glycopeptide-associated tumor vaccines[J]. Journal of China Pharmaceutical University, 2012, 43(2): 97-106.

Cited By

Get Citation

PDF

XML

Article views (237) PDF downloads (481)

Turn off MathJax

Article Contents

Abstract

References

Construction of mutant NDUFS7 plasmid and its effects on neural cells

Abstract

References

Related Articles

Catalog

Construction of mutant NDUFS7 plasmid and its effects on neural cells

Abstract

References

Related Articles

Catalog

Export File

Citation

Format

Content