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Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
Published on: February 22, 2016
Gene expression analysis of the embryonic subplate.
Franziska M Oeschger1, Wei-Zhi Wang, Sheena Lee
1Department of Physiology, Anatomy and Genetics, Oxford University, Oxford OX1 3QX, UK.
Cerebral Cortex (New York, N.Y. : 1991)
|August 25, 2011
Summary
Researchers profiled gene expression in embryonic mouse subplate cells. They identified 13 novel genes crucial for subplate neuron function, axon guidance, and synapse formation.
Area of Science:
- Neuroscience
- Developmental Biology
- Genetics
Background:
- The subplate is a transient embryonic brain layer containing early neurons.
- Subplate cells guide thalamocortical axons and shape cortical circuitry.
- Its cellular heterogeneity and molecular functions remain incompletely understood.
Purpose of the Study:
- To characterize the gene expression profile of the embryonic murine subplate at E15.5.
- To identify novel genes expressed in subplate neurons.
- To investigate the role of these genes in subplate development and function.
Main Methods:
- Laser capture microdissection and microarray analysis of E15.5 murine subplate and cortical plate.
- Quantitative reverse transcription-polymerase chain reaction (RT-qPCR).
- In situ hybridization (ISH) and immunohistochemistry (IHC) for gene validation.
- Analysis in the reeler mutant mouse model.
Main Results:
- Over 300 transcripts showed higher expression in the subplate compared to the cortical plate.
- Confirmed expression of 13 novel genes (e.g., Abca8a, Cdh10, Gabra5) in the E15.5 subplate.
- Expression of 9 of these genes was altered in reeler mutants, correlating with subplate position shifts.
- Identified genes involved in synapse formation, axonal growth, and guidance.
Conclusions:
- The embryonic subplate exhibits a distinct gene expression profile.
- These novel genes are implicated in subplate neuron development and function.
- Subplate gene expression is sensitive to developmental patterning cues, as shown in reeler mutants.
