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The Role Of PCDH? In Hippocampus Axon And Myelin Sheath Development

Posted on:2018-09-12Degree:MasterType:Thesis
Country:ChinaCandidate:W C LuFull Text:PDF
GTID:2404330596991224Subject:Bone science
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Aims: To investigate axon growth and myelination of neurons with pcdh? cluster deletion.To determine the changes of gene transcription of Pcdh? knockout mice hippocampus,especially about axon growth.Methods: The heterozygous Pcdh?-del female mice were crossed with heterozygous Pcdh?-del male mice.one wild-type embryo mouse and one homozygous embryo mouse on an embryonic day 17.5 were prepared for neuron culture.Neurons were staining by immunofluorescent;images were collected with confocal microscopy.The length of axon was analyzed with Image J software and checked manually.The heterozygous Pcdh?-del female mice were crossed with heterozygous Pcdh?-del male mice.one wild-type mouse and one homozygous mouse on postnatal day 21 were prepared for transmission electron microscopy.The samples were photographed using the transmission electron microscope to observation axon myelination.The heterozygous Pcdh?-del female mice were crossed with heterozygous Pcdh?-del male mice.Three wild-type embryo mice and three homozygous embryo mice on an embryonic day 17.5 were prepared for RNA sequencing.RT-q PCR was used to verification the result of the RNA sequencing.Results: Pcdh? knockout results in axon growth defects and fewer myelinated axons.A group of genes involved in axon extension,axon guidance and spinal cord development were down regulated in hippocampus with the absence of Pcdh?.Conclusion: The pcdh? gene cluster may have a potential role in axonal outgrowth and the proper development of myelination.Genes related to the development of CNS especially in axon extension and axon guidance are changed in the absence of Pcdh?.
Keywords/Search Tags:protocadherin ?, axon, myelination, RNA sequencing, hippocampal neuron culture, Transmission electron microscopy
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