Generation of a Tenascin-C-CreER2 Knockin Mouse Line for Conditional DNA Recombination in Renal Medullary Interstitial Cells

Wenjuan He,Chuan-Ming Hao,Matthew D Breyer,Min Zhao,Linda S Davis,Qionghong Xie,Jing Chen,Yingying Wang,Guoqiang Gu,Jeff M Sands

doi:10.1371/journal.pone.0079839

Abstract

Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure. Here, we generated a RMIC specific tenascin-C promoter driven inducible CreER2 knockin mouse line with an EGFP reporter. Similar as endogenous tenascin-C expression, the reporter EGFP expression in the tenascin-C-CreER2+/− mice was observed in the inner medulla of the kidney, and co-localized with COX2 but not with AQP2 or AQP1, suggesting selective expression in RMICs. After recombination (tenascin-C-CreER2+/−/ROSA26-lacZ+/− mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs. Cre activity was not obvious in other major organs or without tamoxifen treatment. This inducible RMIC specific Cre mouse line should therefore provide a novel tool to manipulate genes of interest in RMICs.

Highlights

The renal medullary interstitial cells (RMIC) are a population of specialized stroma-like cells in renal medulla
Animal studies show that chemical ablation of RMICs with BEA leads to systemic hypertension [3]
To better understand the molecular basis of the physiological roles of RMICs, a Cre-recombinase/ LoxP-based RMIC-specific gene deletion could be a powerful approach to investigate the significance of specific genes in RMICs

Summary

Introduction

The renal medullary interstitial cells (RMIC) are a population of specialized stroma-like cells in renal medulla. Prior to hybridization, tissue sections were deparaffinized, refixed in 4% paraformaldehyde, treated with proteinase K (20 mg/ml), washed with PBS, refixed in 4% paraformaldehyde, Figure 1. High levels of tenascin-C mRNA expression in the mouse renal medullary interstitium.

Results

Conclusion