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Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development
Published on: April 3, 2013
Microarray analysis of the developing cortex
Mawahib O Semeralul1, Paul C Boutros, Olga Likhodi
1Department of Pharmacology, Faculty of Medicine, University of Toronto, Ontario, Canada.
Insights
Gene expression in the developing mouse prefrontal cortex (PFC) decreases significantly after birth, particularly between weeks 2 and 4. This study maps these crucial postnatal transcriptomic changes.
Area of Science:
- Neuroscience
- Developmental Biology
- Genomics
Background:
- Abnormal prefrontal cortex (PFC) development is linked to childhood and adolescent-onset neuropsychiatric disorders.
- While the basic PFC structure forms in utero, significant remodeling occurs throughout adolescence.
Purpose of the Study:
- To comprehensively map gene transcript changes in the mouse PFC during postnatal development.
- To identify specific gene functions and pathways affected during this critical period.
Main Methods:
- Serial measurements of messenger RNA (mRNA) levels in mouse PFC using oligonucleotide microarrays.
- Quantitative PCR (qPCR) to validate microarray findings for selected genes.
Main Results:
- Observed significant changes in mRNA transcripts correlating with known postnatal developmental events.
- Most significantly altered transcripts showed a progressive decrease in abundance after birth, with the most substantial changes occurring between postnatal weeks 2 and 4.
- Decreased mRNA levels were noted for genes involved in cell proliferation, cytoskeletal function, extracellular matrix, membrane transport, protein folding, and regulation.
Conclusions:
- Postnatal development of the mouse PFC involves widespread decreases in gene transcript abundance.
- These transcriptomic changes impact fundamental cellular functions and processes critical for PFC maturation.
- Understanding these developmental gene expression patterns is vital for insights into neuropsychiatric disorders with adolescent onset.
Abstract:
Abnormal development of the prefrontal cortex (PFC) is associated with a number of neuropsychiatric disorders that have an onset in childhood or adolescence. Although the basic laminar structure of the PFC is established in utero, extensive remodeling continues into adolescence. To map the overall pattern of changes in cortical gene transcripts during postnatal development, we made serial measurements of mRNA levels in mouse PFC using oligonucleotide microarrays. We observed changes in mRNA transcripts consistent with known postnatal morphological and biochemical events. Overall, most transcripts that changed significantly showed a progressive decrease in abundance after birth, with the majority of change between postnatal weeks 2 and 4. Genes with cell proliferative, cytoskeletal, extracellular matrix, plasma membrane lipid/transport, protein folding, and regulatory functions had decreases in mRNA levels. Quantitative PCR verified the microarray results for six selected genes: DNA methyltransferase 3A (Dnmt3a), procollagen, type III, alpha 1 (Col3a1), solute carrier family 16 (monocarboxylic acid transporters), member 1 (Slc16a1), MARCKS-like 1 (Marcksl1), nidogen 1 (Nid1) and 3-hydroxybutyrate dehydrogenase (heart, mitochondrial) (Bdh).
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