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Updated: Jan 3, 2026
Detection of G Protein-coupled Receptor Expression in Mouse Vagal Afferent Neurons using Multiplex In Situ Hybridization
Published on: September 20, 2021
Distribution pattern of GPRC6A mRNA in mouse tissue by in situ hybridization
Junming Luo1, Zhaoqian Liu, Jinshi Liu
1Kidney Institute, Department of Medicine, University of Kansas, KS 66160, USA. jluo099@yahoo.com
Objective:
To explore the distribution pattern of G protein-coupled receptor family C, group 6, subtype A (GPRC6A) mRNA in adult mice.
Methods:
The distribution of GPRC6A mRNA in paraffin embedded adult mouse tissues was determined by highly sensitive nonradioactive cRNA probe in situ hybridization (ISH). We compared ISH with and without addition of tyramide signal amplification (TSA). GPRC6A wild-type and littermate GPRC6A null mice tissue sections were investigated by ISH.
Results:
TSA greatly increased the sensitivity of ISH to detect GPRC6A mRNA in wild type mouse tissues. There was no detection of GPRC6A mRNA in GPRC6A gene specific knockout tissue in paraffin embedded tissue section. The mRNA of GPRC6A was detectable in the digestive gland or accessory digestive gland including salivary gland and pancreas, as well as in the tissues including kidney, testis, brain, muscle, and fat.
Conclusion:
The mRNA distribution pattern of GPRC6A gene is compatible with the phenotype of GPRC6A knockout mice.
Insights
G protein-coupled receptor family C, group 6, subtype A (GPRC6A) mRNA is found in mouse digestive glands, kidney, testis, brain, muscle, and fat. Its distribution pattern aligns with GPRC6A knockout mouse phenotypes.
Area of Science:
- Molecular Biology
- Genetics
- Physiology
Background:
- G protein-coupled receptor family C, group 6, subtype A (GPRC6A) is a receptor with largely unknown functions.
- Understanding the tissue distribution of GPRC6A mRNA is crucial for elucidating its physiological roles.
Purpose of the Study:
- To map the distribution of GPRC6A mRNA in adult mouse tissues.
- To validate the sensitivity of in situ hybridization (ISH) with tyramide signal amplification (TSA) for GPRC6A mRNA detection.
Main Methods:
- Nonradioactive cRNA probe in situ hybridization (ISH) was employed on paraffin-embedded adult mouse tissues.
- Tyramide signal amplification (TSA) was used to enhance ISH sensitivity.
- GPRC6A wild-type and knockout mouse tissues were analyzed.
Main Results:
- TSA significantly improved the detection sensitivity of GPRC6A mRNA in wild-type mouse tissues.
- No GPRC6A mRNA signal was detected in GPRC6A knockout tissues, confirming assay specificity.
- GPRC6A mRNA was detected in salivary glands, pancreas, kidney, testis, brain, muscle, and adipose tissue.
Conclusions:
- The observed mRNA distribution of GPRC6A is consistent with the known phenotypes of GPRC6A knockout mice.
- These findings provide a foundation for further research into GPRC6A function in various tissues.
