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Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence
Published on: April 17, 2019
Changing expression of chloride channels during preimplantation mouse development
I Arnaiz1, M H Johnson, D I Cook
1School of Medical Sciences, Discipline of Physiology and Bosch Institute, University of Sydney, Sydney, New South Wales 2006, Australia.
Reproduction (Cambridge, England)
|November 2, 2012
Summary
Chloride channels (ClCs) are crucial for mouse embryo development. This study found specific ClC currents essential for blastocyst expansion, highlighting their role in early mammalian development.
Area of Science:
- Developmental Biology
- Cell Physiology
- Molecular Biology
Background:
- Plasma membrane chloride channels (ClCs) regulate vital cellular functions like volume control and transport.
- These functions are critical for successful preimplantation embryonic development.
Purpose of the Study:
- To investigate the molecular and functional expression of voltage-gated ClCs during mouse preimplantation development.
- To determine the role of these channels in conceptus development and blastocyst expansion.
Main Methods:
- Analysis of Clcn gene mRNA expression using RT-PCR.
- Detection of CLCN3 protein localization via immunofluorescence.
- Electrophysiological recordings (whole-cell patch-clamp) to characterize chloride currents.
- Pharmacological inhibition of chloride channels using 9-AC, DIDS.
Main Results:
- All Clcn gene transcripts were detected; Clcn1 mRNA showed differential expression in blastocysts (trophectoderm vs. inner cell mass).
- CLCN3 protein was observed in early embryos and trophoblasts.
- Two distinct chloride currents were identified: a DIDS-sensitive, outwardly rectifying current present throughout development, and a DIDS-insensitive current active during trophoblast differentiation and blastocyst expansion.
- Inhibition of both currents with 9-AC blocked blastocyst expansion.
Conclusions:
- Voltage-gated chloride channels are expressed and functionally active throughout mouse preimplantation development.
- Both DIDS-sensitive and DIDS-insensitive chloride currents play essential roles in blastocyst expansion.
- ClC channels are critical for successful early mammalian development.
