Using RNA sequencing to identify transcriptomic differences in the renal pelvis

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Fox, Emily Brooke

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2023

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Thesis

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peristaltic contraction , renal pelvis , RNA sequencing , single-cell , smooth muscle

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Abstract

The upper urinary tract is an integral part of the pyelocaliceal system, acting as a bridge between the kidney and the bladder. To promote the passage of urine out of the kidney, the renal pelvis generates peristaltic contractions. A specialized population of resident pacemaker cells are required to initiate and maintain peristalsis; however, the precise mechanisms that underlie pacemaker function remain undetermined. By studying gene expression of pacemaker cells, our goal is to gain further insight as to the workings of this tissue. Since genetic studies of the renal pelvis have not been well established, in this thesis we optimized cell and tissue RNA isolation methods to obtain higher yields and high-quality RNA for our studies. We use a combination of transcriptomic approaches including bulk and single-cell RNA sequencing to reveal gene expression in multiple renal pelvis cell types, including pacemaker cells. We also report the generation of a new transgenic reporter line that may improve our ability to distinguish pacemaker cells and discuss improvements that can be implemented in future research. This study provides a strong foundation for upcoming work and signals the exciting beginnings of investigating the genetic makeup of the renal pelvis in health and disease.

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