Coupling of P2Y receptors to Ca2+ mobilization in mesenchymal stromal cells from the human adipose tissue

Abstract

The purinergic transduction was examined in mesenchymal stromal cells (MSCs) from the human adipose tissue, and several nucleotides, including ATP, UTP, and ADP, were found to mobilize cytosolic Ca²⁺. Transcripts for multiple purinoreceptors were detected in MSC preparations, including A₁, A_2A, A_2B, P2Y₁, P2Y₂, P2Y₄, P2Y₆, P2Y₁₁, P2Y₁₃, P2Y₁₄, P2X₂, P2X₄, and P2X₇. Cellular responses to nucleotides were insignificantly sensitive to bath Ca²⁺, pointing at a minor contribution of Ca²⁺ entry, and were suppressed by U73122 and 2-APB, implicating the phosphoinositide cascade in coupling P2Y receptors to Ca²⁺ release. While individual cells were sensitive to several P2Y agonists, responsiveness to a given nucleotide varied from cell to cell, suggesting that particular MSCs could employ different sets of purinoreceptors. Caged Ca²⁺ stimulated Ca²⁺-induced Ca²⁺ release (CICR) that was mediated largely by IP₃ receptors, and resultant Ca²⁺ transients were similar to nucleotide responses by magnitude and kinetics. A variety of findings hinted at CICR to be a universal mechanism that finalizes Ca²⁺ signaling initiated by agonists in MSCs. Individual MSCs responded to nucleotides in an all-or-nothing manner. Presumably just CICR provided invariant Ca²⁺ responses observed in MSCs at different nucleotide concentrations. The effects of isoform specific agonists and antagonists suggested that both P2Y₁ and P2Y₁₃ were obligatory for ADP responses, while P2Y₄ and P2Y₁₁ served as primary UTP and ATP receptors, respectively. Extracellular NAD⁺ stimulated Ca²⁺ signaling in each ATP-responsive MSC by involving P2Y₁₁. The overall data indicate that extracellular nucleotides and NAD⁺ can serve as autocrine/paracrine factors regulating MSC functions.

Keywords: Adipose tissue; Calcium-induced calcium release; IP(3) receptors; Mesenchymal stromal cells; Purinoreceptors.