The James’ lab is focused on identifying and characterizing different proteins that are essential for male fertility. A specific family of proteins called Na+/H+ Exchangers (NHEs) are present in cells, and are known to regulate the internal pH of a cell. NHE10 is one Na+/H+ exchanger known to be expressed exclusively in sperm. Of particular interest, mice deficient for the NHE10 protein are completely infertile. It is also believed that NHE11, which is from the same gene subfamily as NHE10 (SLC9C gene subfamily), is also expressed in sperm, however nothing has been published on the NHE11 protein to date. The goal for this project is to generate two transgenic mice lines which express the rat isoforms of NHE10 and NHE11, then characterize these proteins’ role in sperm physiology. This project consists of assembling DNA vectors which contain the genes that code for either rat NHE10 or NHE11 and then injecting the vectors into mouse embryos to generate transgenic mice. Right now, we have DNA vectors which code for rat NHE10 and NHE11 fully sequenced, and we have digested them in order to isolate the genes of interest. We have injected mice embryos in order to generate these transgenic mice lines and are now waiting for the mice to be born so that we can molecularly characterize them to ensure that the protein of interest is actually expressed. We plan on breeding these transgenic mice into a background of NHE10 deficient mice to determine if rat NHE10/NHE11 are functionally redundant. We expect that the rat NHE11 protein is able to rescue fertility in mice that lack NHE10 expression. Ultimately we are looking to be the first group to characterize the function of NHE11. I intend on pursuing a career as a physician scientist, so this research experience has been incredibly relevant to my future. My work in the James lab has increased my understanding of molecular biology, as well as giving me a basis in general laboratory techniques, literature review, and scientific writing skills.
Author: Jason Abele
Faculty Advisor: Paul James, Biology
Graduate Student Advisor: Cameron Gardner, Biology
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