Super-resolution microscopy localizes perilipin 5 at lipid droplet-mitochondria interaction sites and at lipid droplets juxtaposing to perilipin 2

Research output: Contribution to journalArticleAcademicpeer-review

38 Citations (Web of Science)
43 Downloads (Pure)

Abstract

Objective: Intramyocellular lipid droplets (LD) and their coat proteins PLIN2 and PLIN5 are involved in lipolysis, with a putative role for PLIN5 in mitochondrial tethering. Reportedly, these proteins co-localize and cover the surface of the LD. To provide the spatial basis for understanding how these proteins possess their distinct roles, we examined the precise location of PLIN2 and PLIN5 and explored PLIN5 presence at LD-mitochondria contact sites using Stimulated emission depletion (STED) microscopy and correlative light-electron microscopy (CLEM) in human skeletal muscle sections.

Methods: LDs were stained by MDH together with combinations of mitochondria] proteins and PLINs. Subcellular distribution and co-localization of PLIN proteins and mitochondria was imaged by STED microscopy (Leica TCS SP8) and quantified using Pearson's correlation coefficients and intensity profile plots. CLEM was employed to examine the presence of PLIN5 on mitochondria-LD contact sites.

Results: Both PLIN2 and PLIN5 localized to the LD in a dot-like, juxtaposed fashion rather than colocalizing and covering the entire LD. Both STED and CLEM revealed a high fraction of PLIN5 at the LD-mitochondria interface, but not at mitochondrial cristae, as suggested previously.

Conclusion: Using two super-resolution imaging approaches, this is the first study to show that in sections of human skeletal muscle PLIN2 and PLIN5 localize to the LD at distinct sites, with abundance of PLIN5 at LD-mitochondria tethering sites. This novel spatial information uncovers that PLIN proteins do not serve as lipolytic barriers but rather are docking sites for proteins facilitating selective lipase access under a variety of lipolytic conditions.

Original languageEnglish
Pages (from-to)1423-1432
Number of pages10
JournalBiochimica et Biophysica Acta-Molecular and Cell Biology of Lipids
Volume1863
Issue number11
DOIs
Publication statusPublished - Nov 2018

Keywords

  • Perilipin 5
  • Perilipin 2
  • Lipid droplets
  • Skeletal muscle
  • Mitochondria
  • Super-resolution microscopy
  • ADIPOSE TRIGLYCERIDE LIPASE
  • INDUCED INSULIN-RESISTANCE
  • FLUORESCENCE MICROSCOPY
  • SKELETAL-MUSCLE
  • STIMULATED-EMISSION
  • PROTEIN
  • RESOLUTION
  • SENSITIVITY
  • LIPOLYSIS
  • DYNAMICS

Cite this