Related Experiment Video
Updated: May 15, 2025
Volumetric Imaging and Analysis of Primary Cilia in Musculoskeletal Tissue using the ARL13B-CENTRIN-2 Mouse Model
Published on: March 28, 2025
ARL13B-Cerulean rescues Arl13b-null mouse from embryonic lethality and reveals a role for ARL13B in spermatogenesis
Alyssa B Long1, Isabella M Wilson1,2, Tiffany T Terry1
1Department of Human Genetics, Emory University School of Medicine, 615 Michael Street, Suite 301, Atlanta, GA 30322, USA.
Abstract:
ARL13B is a regulatory GTPase enriched in cilia, making it a popular marker for this organelle. Arl13bhnn/hnn mice lack ARL13B expression, die during midgestation, and exhibit defects in ciliogenesis. The R26Arl13b-Fucci2aR biosensor mouse line directs the expression of fluorescently tagged full-length Arl13b cDNA upon Cre recombination. To determine whether constitutive, ubiquitous expression of ARL13B-Cerulean can replace endogenous gene expression, we generated Arl13bhnn/hnn animals expressing ARL13B-Cerulean. We show that Arl13bhnn/hnn;Arl13b-Cerulean mice survive to adulthood with no obvious physical or behavioral defects, indicating that the fluorescently tagged protein can functionally replace the endogenous protein during development. However, we observed that rescued males failed to sire offspring, revealing a role for ARL13B in spermatogenesis. This work shows that the R26Arl13b-Fucci2aR mouse contains an inducible allele of Arl13b capable of functioning in most tissues and biological processes.
Insights
ARL13B (Arf-like protein 13B) is crucial for cilia development. Rescued mice expressing fluorescent ARL13B survived but showed male infertility, revealing ARL13B
Area of Science:
- Cell Biology
- Genetics
- Developmental Biology
Background:
- ARL13B is a regulatory GTPase vital for cilia formation.
- Mice lacking ARL13B die in midgestation with ciliogenesis defects.
Purpose of the Study:
- To assess if fluorescently tagged ARL13B can functionally replace endogenous ARL13B.
- To investigate the role of ARL13B in spermatogenesis.
Main Methods:
- Generation of Arl13b knockout mice expressing ARL13B-Cerulean.
- Phenotypic analysis of rescued mice, including survival, development, and reproductive capacity.
Main Results:
- Arl13b knockout mice expressing ARL13B-Cerulean survived to adulthood without overt defects.
- Rescued male mice were infertile, indicating a role for ARL13B in spermatogenesis.
- The R26Arl13b-Fucci2aR mouse line provides an inducible Arl13b allele.
Conclusions:
- Fluorescently tagged ARL13B can rescue developmental defects associated with Arl13b deficiency.
- ARL13B plays a critical, previously unrecognized role in male fertility.
- The R26Arl13b-Fucci2aR mouse is a valuable tool for studying Arl13b function.
