Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

HIF-1α/MMP-9 Axis Is Required for the Early Phases of Skeletal Myoblast Differentiation under Normoxia Condition In Vitro

Flaminia Chellini
ORCID logo ,
Alessia Tani
ORCID logo ,
Martina Parigi
,
Francesco Palmieri
ORCID logo ,
Rachele Garella
,
Sandra Zecchi-Orlandini
,
Roberta Squecco
* ORCID logo ,
Chiara Sassoli
*
Version 1 : Received: 7 November 2023 / Approved: 8 November 2023 / Online: 8 November 2023 (10:31:12 CET)

A peer-reviewed article of this Preprint also exists.

Chellini, F.; Tani, A.; Parigi, M.; Palmieri, F.; Garella, R.; Zecchi-Orlandini, S.; Squecco, R.; Sassoli, C. HIF-1α/MMP-9 Axis Is Required in the Early Phases of Skeletal Myoblast Differentiation under Normoxia Condition In Vitro. Cells 2023, 12, 2851. Chellini, F.; Tani, A.; Parigi, M.; Palmieri, F.; Garella, R.; Zecchi-Orlandini, S.; Squecco, R.; Sassoli, C. HIF-1α/MMP-9 Axis Is Required in the Early Phases of Skeletal Myoblast Differentiation under Normoxia Condition In Vitro. Cells 2023, 12, 2851.

Abstract

Hypoxia-inducible factor (HIF)-1α represents the oxygen-sensitive subunit of HIF transcriptional factor, usually degraded in normoxia and stabilized in hypoxia to regulate several target gene expression. Nevertheless, in the skeletal muscle satellite stem cells (SCs), an oxygen level-independent regulation of HIF-1α has been observed. Although HIF-1α has been highlighted as SC function regulator, its spatio-temporal expression and role during myogenic progression remain controversial. We herein analyzed HIF-1α expression and localization in differentiating murine C2C12 myoblasts and SCs under normoxia and evaluated matrix metalloproteinase (MMP)-9 as a HIF-1α target, by biomolecular, biochemical, morphological and electrophysiological analyses. HIF-1α expression increased after 24/48 h of differentiating culture and tended to decline after 72 h/5 days. Committed and proliferating mononuclear myoblasts exhibited nuclear HIF-1α expression, differently from the more elongated and parallel-aligned cells, likely ready to fuse with each other, which show mainly a cytoplasmic localization of the factor. Multinucleated myotubes displayed both a nuclear and cytoplasmic HIF-1α expression. The MMP-9 and MyoD (myogenic activation marker) expression synchronized with that of HIF-1α, increasing after 24 h of differentiation. Short-interfering RNA silencing of HIF-1α and MMP-9 and MMP-9 pharmacological inhibition unraveled MMP-9 as HIF-1α downstream effector and HIF-1α/MMP-9 axis essential for the morpho-functional cell myogenic commitment.

Keywords

electrophysiology; HIF; immunofluorescence; MMP-9; myogenesis; myoblasts; normoxia; regeneration; satellite cells; skeletal muscle.

Subject

Biology and Life Sciences, Cell and Developmental Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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