NDLI: Effects of Lyrm1 knockdown on mitochondrial function in 3 T3-L1 murine adipocytes

Content Provider	Springer Nature Link
Author	Zhu, Guan Zhong Zhang, Min Kou, Chun Zhao Ni, Yu Hui Ji, Chen Bo Cao, Xin Guo Guo, Xi Rong
Copyright Year	2012
Abstract	To explore the effects of Lyrm1 knockdown on the mitochondrial function of 3 T3-L1 adipocytes using small interfering RNA (siRNA). 3 T3-L1 preadipocytes were infected with either a negative control (NC) expression lentivirus or a Lyrm1-shRNA expression lentivirus and induced to differentiate. The knockdown efficiency of Lrym1-specific shRNA in 3 T3-L1 cells was evaluated by real-time PCR. The ultrastructure of the mitochondria in adipocytes was visualized using transmission electron microscopy after differentiation. The levels of mitochondrial DNA copy numbers and Ucp2 mRNA were detected by real-time quantitative PCR. The levels of ATP production was detected using a photon-counting luminometer. The mitochondrial membrane potential and ROS levels of cells were analyzed with a FACScan flow cytometer using Cell Quest software. Cells transfected with lentiviral-Lyrm1-shRNA showed a significantly reduced transcription of Lyrm1 mRNA compared with NC cells. The size and ultrastructure of mitochondria in Lyrm1 knockdown adipocytes was similar to those of the NC cells. There was no significant difference in mtDNA copy number between the two groups. The total level of ATP production, mitochondrial membrane potential and Ucp2 mRNA expression levels were dramatically increased in adipocytes transfected with Lyrm1 RNAi. Furthermore, the level of ROS was dramatically decreased in Lyrm1 knockdown adipocytes. Knockdown of the Lyrm1 gene in adipocytes resulted in dramatically increased cellular ATP production, mitochondrial membrane potentials and levels Ucp2 mRNA, while ROS levels were significantly decreased. These results imply that mitochondrial function is improved in adipocytes after the knockdown of Lyrm1.
Starting Page	225
Ending Page	232
Page Count	8
File Format	PDF
ISSN	0145479X
Journal	Journal of Bioenergetics and Biomembranes
Volume Number	44
Issue Number	1
e-ISSN	15736881
Language	English
Publisher	Springer US
Publisher Date	2012-01-15
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Obesity RNA interference Mitochondria Ucp2 Animal Anatomy / Morphology / Histology Animal Biochemistry Bioorganic Chemistry Organic Chemistry Biochemistry
Content Type	Text
Resource Type	Article
Subject	Cell Biology Physiology

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