| Title: |
The Loss of Drp1 Improves Skeletal Muscle Insulin Action in Primary Myotubes Derived from Humans with Severe Obesity |
| Authors: |
Kugler, Benjamin A.; Labaf, Maryam; Nguyen, Paul; Lin, Nana; Houmard, Joseph A.; Zarringhalam, Kourosh; Zou, Kai |
| Source: |
The FASEB Journal ; volume 36, issue S1 ; ISSN 0892-6638 1530-6860 |
| Publisher Information: |
Wiley |
| Publication Year: |
2022 |
| Collection: |
Wiley Online Library (Open Access Articles via Crossref) |
| Description: |
Dynamin‐related protein 1 (Drp1) is a key regulator of mitochondrial fission. Excessive Drp1‐mediated mitochondrial fission in skeletal muscle from humans with severe obesity is associated with impaired insulin action. However, it remains unclear whether specific inhibition of Drp1 in skeletal muscle cells alleviates insulin resistance in obesity. Therefore, this study aims to determine the direct role of Drp1 on regulating insulin action in human skeletal muscle cells derived from humans with severe obesity. Human skeletal muscle cells from six lean, insulin‐sensitive (LN, BMI = 22.7 ± 1.2 kg/m 2 ,HOMA‐IR = 1.9 ± 0.4) and six severely obese, insulin‐resistant (OB, BMI = 47.3 ± 2.8 kg/m 2 , HOMA‐IR = 3.4 ± 0.4) subjects were pooled together, respectively. At 90% confluency, myoblasts were transfected using polyethyleneimine with a Drp1 shRNA (shDrp1) or scramble shRNA constructs (shCtrl). After 48 h, the medium was replaced with differentiation media with puromycin. On day 7 of differentiation, the mitochondrial network, reactive oxygen species (ROS), insulin signaling, glucose uptake, and protein markers of mitochondrial dynamics and mitochondrial content were assessed. RNA sequencing was also performed on OB‐shCtrl and OB‐shDrp1 myotubes. Differentially regulated genes were identified, and a gene set enrichment was used to determine pathway modulations. Drp1 protein expression was reduced in OB‐shDrp1 myotubes compared to LN‐shCtrl and OB‐shCtrl (72% and 78%, respectively, P |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1096/fasebj.2022.36.s1.r4873 |
| Availability: |
https://doi.org/10.1096/fasebj.2022.36.s1.r4873 |
| Rights: |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
| Accession Number: |
edsbas.3BEACA17 |
| Database: |
BASE |