Purinergic Mechansism of Nociception

描述（由申请人提供）： Cebra的第1阶段申请旨在测试伤害感受感觉神经元亚组中存在的两个ATP门控离子通道的同时功能下调的可行性。 选择P2 X2和P2 X3嘌呤能受体进行研究，是因为最近的研究表明，许多感觉神经元的行为就像它们以异源多聚体构型（P2 X2/3）表达这两种受体，并且因为P2 X3单独的反义寡核苷酸敲低导致一些伤害性行为的适度减弱。 该提议的基本假设是，同时敲除P2 X2和P2 X3嘌呤能受体是干扰伤害性处理的有效方法，并且它是比单独靶向P2 X2或P2 X3受体的方法更有效的靶向异聚体受体构型的方法。 将使用反义寡核苷酸方法结合免疫组织化学、行为和电生理测量来确定同时递送P2 X2和P2 X3的反义寡核苷酸是否会降低感觉神经元中P2 X2和P2 X3的表达并减少伤害性行为。 此外，这些研究将首次确定反义治疗是否会改变具有完整感受野的功能鉴定的感觉神经元的反应特性。 在提出这一建议时，将综合现有的方法，并作为一种新的范例加以应用。 这些实验的结果可能有助于开发可行的治疗替代品，以替代使用具有高耐受性和依赖性潜力的镇痛剂，如阿片类化合物。

DESCRIPTION (provided by applicant): This Stage 1 Cebra application seeks to test the feasibility of simultaneous functional down-regulation of two ATP gated ion channels present in the subset of nociceptive sensory neurons. The P2X2 and P2X3 purinergic receptors arechosen for investigation because recent studies suggest that many sensory neurons behave as if they express these two receptors in a heteromultimeric configuration (P2X2/3), and because antisense oligonucleotide knockdown of P2X3 alone results in modest attenuation of some nociceptive behaviors. The underlying hypotheses of this proposal is that simultaneous knockdown of P2X2 and P2X3 purinergic receptors is an effective approach to interfere with nociceptive processing, and that it is a more efficient way to target heteromeric receptor configurations than approaches targeting either P2X2 or P2X3 receptors alone. An antisense oligonucleotide approach combined with immunohistochemical, behavioral, and electrophysiological measures will be used to determine whether simultaneous delivery of antisense oligonucleotides for P2X2 and P2X3 will decrease the expression of P2X2 and P2X3 in sensory neurons and reduce nociceptive behaviors. In addition, these studies will determine for the first time whether antisense treatment will alter the response properties of functionally identified sensory neurons with intact receptive fields. In making this proposal, existing methodologies will be integrated and applied as a new paradigm. Findings from these experiments could contribute to the development of viable therapeutic alternatives to the use of analgesic agents such as opiate compounds that have high potential for tolerance and dependence.