An immunohistochemical identification key for cell types in adult mouse prostatic and urethral tissue sections.

Kyle A Wegner,Mark T Cadena,Adam Gottschalk,Jinjin Guo,Jill A Mcmahon,Ryan Trevena,Andrew P Mcmahon,Anne E Turco,Richard B Halberg,Chad M Vezina,Mohammad Saleem

doi:10.1371/journal.pone.0188413

Abstract

Though many methods can be used to identify cell types contained in complex tissues, most require cell disaggregation and destroy information about where cells reside in relation to their microenvironment. Here, we describe a polytomous key for cell type identification in intact sections of adult mouse prostate and prostatic urethra. The key is organized as a decision tree and initiates with one round of immunostaining for nerve, epithelial, fibromuscular/hematolymphoid, or vascular associated cells. Cell identities are recursively eliminated by subsequent staining events until the remaining pool of potential cell types can be distinguished by direct comparison to other cells. We validated our identification key using wild type adult mouse prostate and urethra tissue sections and it currently resolves sixteen distinct cell populations which include three nerve fiber types as well as four epithelial, five fibromuscular/hematolymphoid, one nerve-associated, and three vascular-associated cell types. We demonstrate two uses of this novel identification methodology. We first used the identification key to characterize prostate stromal cell type changes in response to constitutive phosphatidylinositide-3-kinase activation in prostate epithelium. We then used the key to map cell lineages in a new reporter mouse strain driven by Wnt10aem1(cre/ERT2)Amc. The identification key facilitates rigorous and reproducible cell identification in prostate tissue sections and can be expanded to resolve additional cell types as new antibodies and other resources become available.

Highlights

The GenitoUrinary Development Molecular Anatomy Project (GUDMAP, www.gudmap.org) is a multi-laboratory consortium dedicated to providing the scientific and medical communities with hypothesis-generating data and tools to facilitate research
Application of the identification key to mouse prostate and prostatic urethra The male mouse genitourinary tract consists of multiple organs and the type and distribution of cells within organs differs across body axes
This is the region where many prostatic ducts drain into the urethra and where proliferative and fibrotic pathologies have been observed in mouse models of benign prostatic hyperplasia and urinary obstruction [10]

Summary

Introduction

The GenitoUrinary Development Molecular Anatomy Project (GUDMAP, www.gudmap.org) is a multi-laboratory consortium dedicated to providing the scientific and medical communities with hypothesis-generating data and tools to facilitate research. The image repository will facilitate mouse strain selection by investigators, critical evaluation of research results by manuscript and grant reviewers, and generally enhance the rigor and reproducibility of cre/lox research studies. The most significant challenge in developing this repository is to accurately assign cre lineage-labels to known genitourinary cell types. A single round of immunostaining is a possible approach for some applications but is insufficient for comprehensive cell identification in complex tissue sections. Cell sorting and single cell RNASeq address the challenge of differentiating closely related cell types in complex tissues, but destroy tissue organization, cell interactions, and information about a cell’s spatial location

Methods

Results

Conclusion