PKC MODULATION OF GS8 AND CALCIUM CHANNEL INTERACTIONS

（申请人摘要）我的博士前和博士后背景包括行为、神经化学、生物化学和分子生物学的研究。博士后期间，在UCHSC药学系Boris Tabakoff博士的指导下，我对G蛋白介导的信号转导产生了特殊的兴趣和专业知识，特别是腺苷酸家族。环化酶。现在我的目标是扩大我在这一领域的专业知识，并获得所需的技能来研究G蛋白如何调节另一类效应物，电压栅钙通道。在计划的培训中，我将学习电生理学，特别是全细胞膜片钳记录。这项技术将使我能够在毫秒-秒的时间尺度上测量G蛋白对通道的调节事件，而不是在腺苷酸环化酶测定的时间尺度上。建议的培训将在ucchsc药学系的William Sather博士（导师）实验室进行。Sather博士的实验室使用常规电生理技术，如全细胞和单通道膜片钳和双电极电压钳技术。共同导师Thomas Dunwiddie博士是另一位电生理学专家。这些实验室和本系提供优良的设施和智力环境。本项目将重点研究蛋白激酶C （PKC）和G蛋白对N-和P/ q型钙通道的调节。Gi/Go偶联受体已被证明以膜分隔的方式抑制参与中枢神经递质释放的N-和P/ q型通道。最近的研究表明，受Gi/ go偶联受体刺激后释放的GBy亚基与抑制通道功能的chafifiel亚基直接相互作用。另一方面，有证据表明PKC的激活可以拮抗GBy对钙通道的膜界抑制。这些研究的目的是详细研究不同PKC同工酶的作用，以及这些同工酶如何调节Gpy与N (alB)-和PIQ （aw）型通道相互作用的机制。这一培训将使我在未来能够以完全独立的研究者身份撰写提案并获得资助，研究不同效应器类的G蛋白调控，这将促进我在学术环境中的职业发展。钙通道在神经元中起着关键作用，并被认为是多种神经系统疾病的靶点，如家族性偏瘫性偏头痛、发作性共济失调I型2和脊髓小脑性共济失调6。

DESCRIPTION: (Applicant's Abstract) My predoctoral and post- doctoral background consists of behavioral, neurochemical, biochemical and molecular biology studies. During my postdoctoral years under the supervision of Dr. Boris Tabakoff, Department of Pharmacology, UCHSC, I developed a special interest and expertise in G protein-mediated signal transduction, especially in the family of adenylyl. cyclases. Now my goal is to expand my expertise in this area and to acquire skills needed to study how G proteins regulate another class of effectors, the voltagegated calcium channels. In the proposed training I will learn electrophysiology, especially whole cell patch clamp recordings. This technique will allow me to measure events of G protein regulation of the channels in the time scale of milliseconds-seconds, as opposed to the time scale of minutes in the adenylyl cyclase enzyme assays. The proposed training will be take place in Dr. William Sather's (Mentor) laboratory at the Deparunent of Pharmacology, UCHSC. Dr. Sather's laboratory uses routinely electrophysiological techniques, such as whole cell and single channel patch clamp and two-electrode voltage clamp techniques. The Co-Mentor, Dr. Thomas Dunwiddie is another expert electrophysiologist. These laboratories and the Department provide excellent facilities and intellectual milieu. The research project will focus on protein kinase C (PKC) and G protein modulation of N- and P/Q-type calcium channels. Gi/Go- coupled receptors have been demonstrated to inhibit, in a membrane- delimited manner, N- and P/Q-type channels that are involved in neurotransmitter release in the CNS. Recent studies have shown that GBy subunits which are released upon Gi/Go-coupled receptor stimulation are directly interacting with the chafifiel al subunits inhibiting the channel function. On the other hand, there is evidence that activation of PKC antagonizes the membrane-delimited inhibition of calcium channels by GBy. The aim of the proposed studies is to investigate in detail the role of different PKC isozymes and the mechanisms how these isozyntes modulate the interactions of Gpy with N (alB)- and PIQ (aW-type channels. This training will allow me in the future to write proposals and receive funding for studies on G protein regulation of different effector classes as a fully independent investigator, which will advance my career in the academic environment. Calcium channels play a key role in neurons and have been implicated as targets of diverse neurological diseases, such as familial herniplegic migraine, episodic ataxia I type 2 and spinocerebellar ataxia 6.