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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Updated: Jul 16, 2026

Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing

Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing

Published on: September 20, 2021

Sex-specific proximal tubular cell differentiation pathways identified by single-nucleus RNA sequencing.

Yueh-An Lu^1,2,3, Tanya Smith^1,2, Sumukh Deshpande^1,2

¹Division of Infection and Immunity, School of Medicine, Cardiff University, Cardiff, UK.

Scientific Reports

|October 14, 2024

Summary

Postnatal kidney growth relies on tubular cell proliferation, not new nephrons. This study reveals proximal tubular cell (PTC) differentiation pathways leading to sex-specific kidney phenotypes, offering therapeutic targets for kidney development and injury.

Keywords:

Cell biology and structure Chronic kidney disease Epithelial Kidney tubule MRNA Proximal tubule Renal epithelial cell Renal fibrosis Renal tubular epithelial cells

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Area of Science:

Developmental Biology
Genomics
Nephrology

Background:

Postnatal kidney growth primarily involves tubule expansion, not nephrogenesis.
The molecular mechanisms driving tubule elongation and cell specialization remain unclear.

Purpose of the Study:

To characterize mouse kidney development at single-cell resolution.
To identify proliferation and differentiation pathways in kidney tubular cells.

Main Methods:

Single nuclear RNA sequencing (snRNA-seq) of mouse kidneys across developmental time points.
Unbiased clustering and gene expression analysis of ~68,775 nuclei.
Trajectory and RNA Velocity analyses to map cell differentiation.

Main Results:

Identified 31 distinct kidney cell clusters, with high proliferation in proximal tubular cells (PTCs) early in development.
Discovered sex-specific gene expression profiles in PTCs at later developmental stages.
Delineated PTC differentiation pathways leading to specialized, sex-specific phenotypes.

Conclusions:

Tubular cell proliferation is crucial for postnatal kidney growth.
PTC differentiation pathways contribute to sex-specific kidney development.
The generated dataset provides insights into kidney development and potential therapeutic targets for kidney injury.