Differentiation of adipose-derived stem cells into Schwann cell phenotype induces expression of P2X receptors that control cell death.
Differentiation of adipose-derived stem cells into Schwann cell phenotype induces expression of P2X receptors that control cell death.
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DOI：
10.1038/cddis.2013.268
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发表时间：
2013-07-25
影响因子：
9
通讯作者：
中科院分区：
文献类型：
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雪旺细胞是周围神经系统发育、髓鞘形成和再生的基础细胞。缓慢的生长速度和收获困难限制了干细胞在再生医学中的应用。一些分子，包括神经类固醇受体和神经递质，被认为与调节干细胞的生理和再生潜力有关。脂肪干细胞可分化为干细胞样表型(DASC)，具有与SC相同的形态和功能特性，是一种有效的SC替代方法。我们以前已经证明，dasc表达γ-氨基丁酸受体，该受体调节其增殖和神经营养潜力，尽管对其他神经递质在asc中的作用知之甚少。在本研究中，我们研究了嘌呤能受体在DASC中的表达。应用逆转录聚合酶链式反应(RT-PCR)、免疫印迹分析和免疫细胞化学方法，我们证实了ASCs表达P2X3、P2X4和P2X7嘌呤受体。ASCs向神经胶质表型分化伴随着P2X4和P2X7受体的上调。利用钙离子成像技术，我们已经证明了用三磷酸腺苷(ATP)刺激嘌呤受体可以触发细胞内的钙信号，表明这些受体的功能活动。全细胞电压钳记录显示，ATP和BzATP产生的离子电流可被特定的P2X7拮抗剂完全抑制。最后，利用细胞毒性分析，我们已经证明细胞内钙离子的增加导致dASC死亡，这一效应可以用特定的P2X7拮抗剂来阻止。综上所述，这些结果首次表明，功能性的P2X7受体在dASC中的存在及其与细胞死亡和生存等关键生理过程的联系。DASC中这些新的药理靶点的存在可能为利用DASC进行神经修复的组织工程方法中的细胞存活和神经营养潜力的管理开辟新的机会。
Schwann cells (SCs) are fundamental for development, myelination and regeneration in the peripheral nervous system. Slow growth rate and difficulties in harvesting limit SC applications in regenerative medicine. Several molecules, including receptors for neurosteroids and neurotransmitters, have been suggested to be implicated in regulating physiology and regenerative potential of SCs. Adipose-derived stem cells (ASCs) can be differentiated into SC-like phenotype (dASC) sharing morphological and functional properties with SC, thus representing a valid SC alternative. We have previously shown that dASC express γ-aminobutyric-acid receptors, which modulate their proliferation and neurotrophic potential, although little is known about the role of other neurotransmitters in ASC. In this study, we investigated the expression of purinergic receptors in dASC. Using reverse transriptase (RT)-PCR, western blot analyses and immunocytochemistry, we have demonstrated that ASCs express P2X3, P2X4 and P2X7 purinoceptors. Differentiation of ASCs towards glial phenotype was accompanied by upregulation of P2X4 and P2X7 receptors. Using Ca2+-imaging techniques, we have shown that stimulation of purinoceptors with adenosine 5′-triphosphate (ATP) triggers intracellular Ca2+ signals, indicating functional activity of these receptors. Whole-cell voltage clamp recordings showed that ATP and BzATP induced ion currents that can be fully inhibited with specific P2X7 antagonists. Finally, using cytotoxicity assays we have shown that the increase of intracellular Ca2+ leads to dASC death, an effect that can be prevented using a specific P2X7 antagonist. Altogether, these results show, for the first time, the presence of functional P2X7 receptors in dASC and their link with critical physiological processes such as cell death and survival. The presence of these novel pharmacological targets in dASC might open new opportunities for the management of cell survival and neurotrophic potential in tissue engineering approaches using dASC for nerve repair.
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影响因子：
9
作者：
通讯作者：
--
影响因子：
3.1
作者：
Faroni, Alessandro;Terenghi, Giorgio;Magnaghi, Valerio
通讯作者：
Magnaghi, Valerio
影响因子：
2.7
作者：
Mantovani, Cristina;Mahay, Daljeet;Wiberg, Mikael
通讯作者：
Wiberg, Mikael
影响因子：
6.2
作者：
Kaewkhaw, Rossukon;Scutt, Andy M.;Haycock, John W.
通讯作者：
Haycock, John W.
影响因子：
3.3
作者：
Adams, A. M.;Arruda, E. M.;Larkin, L. M.
通讯作者：
Larkin, L. M.