Faculty, Staff and Student Publications

Publication Date

1-1-2026

Journal

Developmental Biology

DOI

10.1016/j.ydbio.2025.10.017

PMID

41130332

PMCID

PMC12616890

PubMedCentral® Posted Date

11-15-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

Hybridization chain reaction RNA-fluorescent in situ hybridization (HCR RNA-FISH) is a powerful and increasingly used method for visualizing gene expression in cells and tissues. A probe set against polyadenylated RNA (poly(A)) is often used as a positive control for RNA integrity and staining quality. While optimizing this technique in the ovary of the brown anole lizard (Anolis sagrei), we found that the poly(A) probe produced a strikingly specific and intense signal in pyriform cells, a specialized lizard-specific nurse cell type. This staining pattern was found in both whole-mount samples and paraffin sections, suggesting that poly(A) signal intensity can serve as a robust molecular marker for this cell type. The specific and robust signal facilitated segmentation of volumetric data to create the first 3D models of pyriform cells. We also observed unusually diffuse DAPI staining in pyriform cell nuclei, distinguishing them from surrounding granulosa cells, pointing to possible differences in chromatin structure or nuclear organization. Together, these findings highlight the potential of poly(A) probes used in HCR RNA-FISH not only as a technical control, but also as a tool to selectively label specific cell types with high transcriptional activity or storage of abundant poly(A) transcripts.

Keywords

Animals, Female, Lizards, In Situ Hybridization, Fluorescence, Ovary, RNA Probes, Poly A, Lizard, ovary, follicle, granulosa cell, pyriform cell, HCR-FISH

Published Open-Access

yes

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.