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Mast cells express chloride channels of the ClC family
M Kulka1, A Schwingshackl, A D Befus
1Department of Medicine, University of Alberta, Edmonton, Canada.
Objective And Design:
The expression of specific Cl- channels in mast cells (MC) is poorly understood. Because the largest and most ubiquitously expressed family of Cl- channels is the ClC, we studied MC mRNA and protein expression for members of the ClC family.
Methods:
Specific primers were designed to ClC-1 to -7 and reverse-transcriptase polymerase chain reaction (RT-PCR) was performed on RNA from highly purified rat peritoneal mast cells (PMC) and the rat cultured mast cell line (RCMC). Protein expression of ClC-2, -3 and -7 chloride channels in PMC and RCMC was studied using western blotting.
Results:
RT-PCR showed that RCMC expressed mRNA for the Cl- channels (ClC)-2, -3, -4, -5 and -7, while PMC expressed ClC-7. Using Western Blotting, we found protein expression of ClC-2 and ClC-7, but not ClC-3, in RCMC, whereas in PMC neither of these proteins could be detected.
Conclusion:
These results indicate that MC express several members of the ClC family and that these Cl- channels might be important in MC functions.
Insights
Mast cells (MC) express several ClC chloride channels, including ClC-2 and ClC-7. These findings suggest that ClC channels play a role in mast cell functions.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- The role of specific chloride (Cl-) channels in mast cells (MC) remains largely unknown.
- The ClC family represents the largest and most widespread group of Cl- channels.
Purpose of the Study:
- To investigate the mRNA and protein expression of ClC family members in rat peritoneal mast cells (PMC) and a rat cultured mast cell line (RCMC).
Main Methods:
- Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to detect mRNA for ClC-1 to -7.
- Western blotting was employed to assess the protein expression of ClC-2, -3, and -7.
Main Results:
- RT-PCR detected mRNA for ClC-2, -3, -4, -5, and -7 in RCMC, and ClC-7 in PMC.
- Western blotting revealed protein expression of ClC-2 and ClC-7 in RCMC, but not ClC-3. No detectable protein expression of these channels was found in PMC.
Conclusions:
- Mast cells express multiple ClC family chloride channels.
- These identified ClC channels may be significant for mast cell functionality.