ClC-c regulates the proliferation of intestinal stem cells via the EGFR signalling pathway in Drosophila
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Updated: Oct 8, 2025
Isolating Intestinal Stem Cells from Adult Drosophila Midguts by FACS to Study Stem Cell Behavior During Aging
Published on: December 16, 2014
Jinping Huang1, Xiao Sheng2, Zhangpeng Zhuo1
1Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong, China.
Objectives:
Adult stem cells uphold a delicate balance between quiescent and active states, which is crucial for tissue homeostasis. Whereas many signalling pathways that regulate epithelial stem cells have been reported, many regulators remain unidentified.
Materials And Methods:
Flies were used to generate tissue-specific gene knockdown and gene knockout. qRT-PCR was used to assess the relative mRNA levels. Immunofluorescence was used to determine protein localization and expression patterns. Clonal analyses were used to observe the phenotype. RNA-seq was used to screen downstream mechanisms.
Results:
Here, we report a member of the chloride channel family, ClC-c, which is specifically expressed in Drosophila intestinal stem/progenitor cells and regulates intestinal stem cell (ISC) proliferation under physiological conditions and upon tissue damage. Mechanistically, we found that the ISC loss induced by the depletion of ClC-c in intestinal stem/progenitor cells is due to inhibition of the EGFR signalling pathway.
Conclusion:
Our findings reveal an ISC-specific function of ClC-c in regulating stem cell maintenance and proliferation, thereby providing new insights into the functional links among the chloride channel family, ISC proliferation and tissue homeostasis.
Insights
Chloride channel ClC-c regulates intestinal stem cell (ISC) proliferation in Drosophila. Its depletion inhibits the EGFR pathway, impacting tissue homeostasis and stem cell maintenance.
Area of Science:
- Cell Biology
- Stem Cell Biology
- Physiology
Background:
- Adult stem cells maintain tissue homeostasis through a balance of quiescence and activity.
- While many regulators of epithelial stem cells are known, numerous others remain undiscovered.
Purpose of the Study:
- To identify novel regulators of intestinal stem cell (ISC) function.
- To investigate the role of the chloride channel family in ISC maintenance and proliferation.
Main Methods:
- Drosophila melanogaster as a model organism for tissue-specific gene manipulation (knockdown/knockout).
- Quantitative reverse transcription PCR (qRT-PCR) for mRNA level assessment.
- Immunofluorescence for protein localization and expression.
- Clonal analysis for phenotypic observation.
- RNA sequencing (RNA-seq) for downstream mechanism screening.
Main Results:
- ClC-c, a chloride channel, is specifically expressed in Drosophila ISCs/progenitor cells.
- ClC-c regulates ISC proliferation under both physiological conditions and after tissue damage.
- Depletion of ClC-c leads to ISC loss, primarily through the inhibition of the Epidermal Growth Factor Receptor (EGFR) signaling pathway.
Conclusions:
- ClC-c plays a crucial ISC-specific role in maintaining stem cell populations and regulating proliferation.
- These findings highlight a novel link between chloride channels, ISC activity, and tissue homeostasis.
- The study provides new insights into the molecular mechanisms governing stem cell function and tissue repair.
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