Abstract | AIMS: METHODS: Preadipocytes were isolated from interscapular brown adipose tissue (BAT) of Agpat2(-/-) and Agpat2(+/+) newborn mice and cultured/differentiated in vitro. Intracellular lipids were quantified by oil red O staining. Cell death was assessed by lactate dehydrogenase (LDH) activity. Apoptosis and autophagy regulatory factors were determined at the mRNA and protein level with Real-time PCR, immunoblot and immunofluorescence. RESULTS: Adipogenically induced Agpat2(-/-) preadipocytes had fewer lipid-loaded cells and lower levels of adipocyte markers than wild type preadipocytes. Before adipogenic differentiation, autophagy-related proteins (ATGs) ATG3, ATG5-ATG12 complex, ATG7 and LC3II were increased but autophagic flux was reduced, as suggested by increased p62 levels, in Agpat2(-/-) preadipocytes. Adipogenic induction increased LDH levels in the culture media in Agpat2(-/-) preadipocytes but no differences were observed in the activation of Caspase 3 or in markers of autophagic flux. CONCLUSIONS: AGPAT2 is required for in vitro adipogenesis of mouse preadipocytes. Autophagy defects or apoptosis are not involved in the adipogenic failure of Agpat2(-/-) preadipocytes.
|
Authors | Marta Fernández-Galilea, Pablo Tapia, Kelly Cautivo, Eugenia Morselli, Víctor A Cortés |
Journal | Biochemical and biophysical research communications
(Biochem Biophys Res Commun)
Vol. 467
Issue 1
Pg. 39-45
(Nov 06 2015)
ISSN: 1090-2104 [Electronic] United States |
PMID | 26417690
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Copyright | Copyright © 2015 Elsevier Inc. All rights reserved. |
Chemical References |
- Acyltransferases
- 2-acylglycerophosphate acyltransferase
|
Topics |
- Acyltransferases
(deficiency, genetics)
- Adipocytes, Brown
(cytology, enzymology)
- Adipogenesis
(genetics, physiology)
- Animals
- Apoptosis
- Autophagy
- Cell Differentiation
- Cells, Cultured
- Lipid Metabolism
- Mice
- Mice, Knockout
- Stem Cells
(cytology, metabolism)
|