Principal Investigator: Peter Piermarini
Contact Information: E-mail: pmp26@cornell.edu - Phone: 607-253-3473
Sponsor: NIH-NIDDK
Grant Number: 3 K01 DK080194-02S1
Title: ARRA: An Epithelial Model for V-Type H+ - ATPase-driven acid-base Transport
Annual Direct Cost: $50,000
Project Period: 09/30/09-09/29/11
DESCRIPTION (provided by applicant): In the spirit of the American Recovery and Reinvestment Act (ARRA) of 2009, this supplemental request aims to create a job for a research technician who will accelerate the tempo of research of the parent grant (1-K01-DK080194-01). The parent grant is exploring the mechanisms of acid-base transport in the Malpighian (renal) tubules of the yellow-fever mosquito Aedes aegypti to determine if these tubules represent a valid model system for understanding V-type H+-ATPase-driven acid secretion in α-intercalated cells of the mammalian, renal collecting tubule. The proposed supplemental research is important to Aim 3 of the parent grant, which aims to identify,localize, and characterize an ortholog of a mammalian K,Cl cotransporter (KCC) from Aedes Malpighian tubules. This KCC plays a critical role in acid secretion by α-intercalated cells of mammalian renal tubules,but its function in Aedes tubules is unknown. In the proposed supplemental study, a technician will clone the cDNA encoding Aedes KCC from Malpighian tubules. Once an antibody is developed, the technician will localize the expression of Aedes-KCC immunoreactivity in the Malpighian tubule using immunohistochemistry. Lastly, the technician will heterologously express the Aedes KCC in Xenopus oocytes to characterize its function. The principal investigator (Dr. Peter M. Piermarini)—who has extensive experience in these techniques—will train and supervise the technician. By employing a technician to focus on the proposed supplemental research, I will be able to focus on learning new techniques proposed in Aim 4 of the parent grant such as (i) in vitro microperfusion, fluid secretion assays, and electron probe analysis from my primary mentor (Prof. Klaus W. Beyenbach), and (ii) genetic disruption via RNA interference from my co-mentor (Dean Michael I. Kotlikoff). Thus, the supplemental request will (i) create a job, (ii) accelerate the tempo of scientific research, and (iii) enhance the career development of the principal investigator.