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Purinergic signalling is required for calcium oscillations in migratory chondrogenic progenitor cells

  • Signaling and cell physiology
  • Published:
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Abstract

Osteoarthritis (OA) is the most common form of chronic musculoskeletal disorders. A migratory stem cell population termed chondrogenic progenitor cells (CPC) with in vitro chondrogenic potential was previously isolated from OA cartilage. Since intracellular Ca2+ signalling is an important regulator of chondrogenesis, we aimed to provide a detailed understanding of the Ca2+ homeostasis of CPCs. In this work, CPCs immortalised by lentiviral administration of the human telomerase reverse transcriptase (hTERT) and grown in monolayer cultures were studied. Expressions of all three IP3Rs were confirmed, but no RyR subtypes were detected. Ca2+ oscillations observed in CPCs were predominantly dependent on Ca2+ release and store replenishment via store-operated Ca2+ entry; CPCs express both STIM1 and Orai1 proteins. Expressions of adenosine receptor mRNAs were verified, and adenosine elicited Ca2+ transients. Various P2 receptor subtypes were identified; P2Y1 can bind ADP; P2Y4 is targeted by UTP; and ATP may evoke Ca2+ transients via detected P2X subtypes, as well as P2Y1 and P2Y2. Enzymatic breakdown of extracellular nucleotides by apyrase completely abrogated Ca2+ oscillations, suggesting that an autocrine/paracrine purinergic mechanism may drive Ca2+ oscillations in these cells. As CPCs possess a broad spectrum of functional molecular elements of Ca2+ signalling, Ca2+-dependent regulatory mechanisms can be supposed to influence their differentiation potential.

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Abbreviations

2-APB:

2-Aminoethoxydiphenyl borate

ARC:

Arachidonic acid-regulated Ca2+ channel

CPA:

Cyclopiazonic acid

CPC:

Chondrogenic progenitor cell

CRAC:

Calcium release activated Ca2+ channel

ESC:

Embryonic stem cell

FTHM:

Full time at half maximum

HDC:

High density cell culture

hTERT:

Human telomerase reverse transcriptase

IP3R:

Inositol 1,2,3-trisphosphate receptor

MSC:

Mesenchymal stem cell

NCX:

Sodium calcium exchanger

OA:

Osteoarthritis

PBST:

Phosphate-buffered saline with Tween 20

PG:

Proteoglycan

PMCA:

Plasma membrane Ca2+-ATPase

RyR:

Ryanodine receptor

SDS–PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

SERCA:

Sarcoplasmic/endoplasmic reticulum Ca2+-ATPase

SOCE:

Store-operated Ca2+ entry

STIM:

Stromal interacting molecule

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Acknowledgments

We are grateful to Mrs. Krisztina Bíró and Mrs. Róza Őri for the excellent and skillful technical assistance. This work was supported by grants from the Hungarian Science Research Fund (OTKA CNK80709, OTKA NN-107765), from TÁMOP-4.2.2.A-11/1/KONV-2012-0036 and TÁMOP-4.2.2/B-10/1-2010-0024 projects, co-financed by the European Union and the European Social Fund. N.M. was supported by the German Research Foundation (Mi 573/10-1). C.M. was supported by a Mecenatura grant (DEOEC Mec-9/2011) from the Medical and Health Science Centre, University of Debrecen, Hungary; a Short-Term Fellowship from the Federation of European Biochemical Societies (FEBS); and also from the European Union through a Marie Curie Intra-European Fellowship for career development (project number: 625746; acronym: CHONDRION; FP7-PEOPLE-2013-IEF). This paper was supported by the János Bólyai Research Scholarship of the Hungarian Academy of Sciences. T.J. is supported by a Magyary Zoltán postdoctoral fellowship through the project entitled “National Excellence Program—Elaboration and implementation of a national student and researcher supporting system for the convergence region” (TÁMOP 4.2.4.A/2-11-1-2012-0001), co-financed by the Hungarian State, the European Union, and the European Social Fund.

Conflict of interest

All authors disclose that there are neither any financial nor any personal relationships with other people or organisations that could inappropriately influence (bias) their work. There are no conflicts of interests.

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Authors and Affiliations

  1. Department of Anatomy, Histology and Embryology, Faculty of Medicine, University of Debrecen, Nagyerdei krt. 98, H-4032, Debrecen, Hungary

    Csaba Matta, Roland Takács, Tamás Juhász & Róza Zákány

  2. Department of Physiology, Faculty of Medicine, University of Debrecen, Nagyerdei krt. 98, H-4032, Debrecen, Hungary

    János Fodor, Adrienn Tóth & László Csernoch

  3. Research Group for Oral Biology and Tissue Regeneration, Faculty of Medicine, Department of Prosthodontics, Georg August University, 37075, Goettingen, Germany

    Nicolai Miosge

  4. Department of Orthopaedics, Faculty of Medicine, University of Debrecen, Nagyerdei krt. 98, H-4032, Debrecen, Hungary

    Henrik Rybaltovszki

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  1. Csaba Matta
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  2. János Fodor
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Correspondence to Róza Zákány.

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Csaba Matta and János Fodor contributed equally to the work.

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Matta, C., Fodor, J., Miosge, N. et al. Purinergic signalling is required for calcium oscillations in migratory chondrogenic progenitor cells. Pflugers Arch - Eur J Physiol 467, 429–442 (2015). https://doi.org/10.1007/s00424-014-1529-8

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