Expression of KCC4 in distal colon surface epithelia

描述(由申请人提供)：这项建议的长期目标是了解KCI联合转运体KCC4在远端结肠K重吸收的激素和饮食控制中的作用，以及它如何用于调节全身K动态平衡。一种针对大鼠KCC4的抗体将被鉴定并用于研究这种转运蛋白在大鼠结肠近端至远端轴的细胞表达。该建议的具体目的包括：1)充分鉴定和优化抗KCC4抗体，以最大限度地提高其在大鼠体内检测和定位KCC4表达的有效性；2)进行实时聚合酶链式反应、免疫印迹和免疫定位研究，以检测KCC4在大鼠远端结肠上皮细胞中的表达；以及3)比较饮食限制K和Na对KCC4、KCC1和结肠H/K-ATPase表达的影响。这些技术将被带回Flows总部校园，这是一家为西班牙裔服务的机构，在那里它们将用于培训对卫生专业和/或生物医学研究职业感兴趣的(少数族裔)学生。预计这位研究员和赞助商之间将建立长期的合作关系，进一步帮助跟踪这些学生在一家主要的博士授予机构的研究生涯。

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to understand the role of the K+CI- co-transporter, KCC4, in hormonal and dietary control of distal colon K+ reabsorption and how it is used regulating whole body K+ homeostasis. An antibody against rat KCC4 will be characterized and used to study cellular expression of this transporter along the proximal to distal axis of rat colon. The specific aims of this proposal include: 1) fully characterize and optimize an anti-KCC4 antibody to maximize its effectiveness in detecting and localizing KCC4 expression in rat; 2) perform real time PCR, Western blotting and immunolocalization studies to examine expression of KCC4 mRNA and protein in rat distal colon epithelia; and 3) compare effects of dietary restriction of K+ and Na+ on expression of KCC4, KCC1 and colonic H/K-ATPase. These techniques will be brought back to the Fellows home campus, an hispanic-serving institution, where they will be used to train (minority) students who are interested in careers in the health professions and/or biomedical research. It is anticipated that a long-term collaboration will be established between the Fellow and the Sponsor, further helping to track these students to research careers at a major doctoral granting institution.