Regulators of autophagosome formation in Drosophila muscles.

PLoS genetics (2015-02-19)
Jonathan Zirin, Joppe Nieuwenhuis, Anastasia Samsonova, Rong Tao, Norbert Perrimon
要旨

Given the diversity of autophagy targets and regulation, it is important to characterize autophagy in various cell types and conditions. We used a primary myocyte cell culture system to assay the role of putative autophagy regulators in the specific context of skeletal muscle. By treating the cultures with rapamycin (Rap) and chloroquine (CQ) we induced an autophagic response, fully suppressible by knockdown of core ATG genes. We screened D. melanogaster orthologs of a previously reported mammalian autophagy protein-protein interaction network, identifying several proteins required for autophagosome formation in muscle cells, including orthologs of the Rab regulators RabGap1 and Rab3Gap1. The screen also highlighted the critical roles of the proteasome and glycogen metabolism in regulating autophagy. Specifically, sustained proteasome inhibition inhibited autophagosome formation both in primary culture and larval skeletal muscle, even though autophagy normally acts to suppress ubiquitin aggregate formation in these tissues. In addition, analyses of glycogen metabolic genes in both primary cultured and larval muscles indicated that glycogen storage enhances the autophagic response to starvation, an important insight given the link between glycogen storage disorders, autophagy, and muscle function.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
グリセロール, for molecular biology, ≥99.0%
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
グリセロール, ≥99.5%
Sigma-Aldrich
グリセロール, ACS reagent, ≥99.5%
Sigma-Aldrich
グリセロール, ReagentPlus®, ≥99.0% (GC)
Sigma-Aldrich
クロロキン 二リン酸塩, powder or crystals, 98.5-101.0% (EP)
Sigma-Aldrich
グリセロール, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)
Supelco
グリセリン, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
抗α-チューブリン抗体, マウスモノクローナル, clone DM1A, purified from hybridoma cell culture
Sigma-Aldrich
グリセリン, meets USP testing specifications
Sigma-Aldrich
グリセロール, BioXtra, ≥99% (GC)
Sigma-Aldrich
グリセロール 溶液, 83.5-89.5% (T)
Sigma-Aldrich
グリセロール, FCC, FG
Sigma-Aldrich
グリセロール, BioUltra, for molecular biology, anhydrous, ≥99.5% (GC)
Sigma-Aldrich
チラミン, ≥98.0%
USP
グリセリン, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
グリセロール, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E422, anhydrous, 99.0-101.0% (alkalimetric)
Supelco
グリセロール, analytical standard
Supelco
リン酸クロロキン, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
グリセロール, puriss. p.a., ACS reagent, anhydrous, dist., ≥99.5% (GC)
Sigma-Aldrich
グリセロール 溶液, puriss., meets analytical specification of Ph. Eur., BP, 84-88%
Sigma-Aldrich
グリセロール, tested according to Ph. Eur., anhydrous
Supelco
チラミン, analytical standard
Sigma-Aldrich
グリセロール, JIS special grade, ≥99.0%
Sigma-Aldrich
グリセロール, SAJ first grade, ≥98.0%
Sigma-Aldrich
チラミン, 98%, FG
Supelco
4-tert-オクチルフェノールモノエトキシラート 溶液, 10 μg/mL in acetone, analytical standard
Sigma-Aldrich
グリセロール 溶液, puriss. p.a., 86-89% (T)
Supelco
4-tert-オクチルフェノールモノエトキシラート 溶液, 1 μg/mL in acetone, analytical standard