A Flanking Gene Problem Leads to the Discovery of a Gprc5b Splice Variant Predominantly Expressed in C57Bl/6J Mouse Brain and in Maturing Neurons
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BACKGROUND: Gprc5b, a retinoic acid-inducible orphan G protein-coupled receptor (GPCR), is a member of the group C metabotropic glutamate receptor family proteins possibly involved in non-canonical Wnt signaling. Many GPCR transcripts are alternatively spliced, which diversifies this class of proteins in their cell- and tissue-specific signaling, regulatory and/or pharmacological properties. We previously generated p97FE65 isoform-specific knockout mice that showed learning/memory deficits. In this study, we further characterized the 97FE65 null mice using cDNA microarray and RT-PCR analyses. METHODOLOGY/PRINCIPAL FINDINGS: We discovered a novel brain-specific C-terminal splice variant of Gprc5b, Gprc5b_v2, which was differentially expressed in p97FE65 wild type and null mouse brains. The null mice were generated in 129/Sv ES cells, and backcrossed to C57Bl/6J for ten generations. We found that expression of Gprc5b_v2 mRNA in the brains of p97FE65 null mice was dramatically down-regulated (more than 20 fold) compared to their wild type littermates. However, expression profiles of Gprc5b variants and SNP analysis surrounding the FE65 locus suggest that the down-regulation is unlikely due to the altered FE65 function, but rather is caused by gene retention from the 129/Sv ES cells. Consistently, in contrast to ubiquitously expressed Gprc5b_v1, Gprc5b_v2 was predominantly expressed in the brain tissues of C57Bl/6J mice. The alternative splicing of the 3' terminal exon also altered the protein coding sequences, giving rise to the characteristic C-termini. Levels of Gprc5b_v2 mRNA were increased during neuronal maturation, paralleling the expression of synaptic proteins. Overexpression of both Gprc5b variants stimulated neurite-like outgrowth in a neuroblastoma cell line. CONCLUSIONS/SIGNIFICANCE: Our results suggest that Gprc5b-v2 may play a role during brain maturation and in matured brain, possibly through the regulation of neuronal morphology and protein-protein interaction. This study also highlights the fact that unexpected gene retention following repeated backcrosses can lead to important biological consequences.
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GPRC5B is a regulator of neurogenesis and inflammatory signaling in obesity.
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- Authors
- Cool, B. H., Chan, G. C.-K., Lee, L., Oshima, J., Martin, G. M., & Hu, Q.
- Affiliations
- University of Washington (US)
- Corresponding author
- Martin, George M.
- Hu, Qubai
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2010-04-26
- Volume
- 5
- Issue
- 4
- Pages
- e10351
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC2859937
- OpenAlex
- W1979847069
- Cited by
- 18
- References
- 45
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 1 · 2019: 3 · 2018: 2 · 2016: 1 · 2014: 2 · 2013: 1 · 2012: 1
- FWCI
- 0.65
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Funders
- National Institutes of Health (AG-00057)
- National Institute on Aging (T32 AG-00057, R01 AG19711, T32AG000057, R01AG019711)
- National Institute of Neurological Disorders and Stroke (R01NS020498, NS20498)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- GPRC5B, alternative splicing, gene retention, neuronal maturation, brain-specific expression, neurite outgrowth, orphan G protein-coupled receptor, C57BL/6J mice
- MeSH terms
- Animals, Brain, Brain Chemistry, Inbreeding, Mice, Inbred C57BL, Neurons, RNA, Messenger, Mice, Knockout, Protein Isoforms, Polymorphism, Single Nucleotide, Gene Expression Profiling, Receptors, G-Protein-Coupled, Mice
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