Expression Profile of Primary Microcephaly GenesCdk5rap2,Cep63and Their ParaloguesPde4dip,Ccdc67in Mouse Embryonic and Adult Tissues

Abstract

AbstractAutosomal recessive primary microcephaly (MCPH) is characterized by the prenatal reduction in the human brain growth, without any change in the cerebral architecture. MCPH can result from the bi-allelic mutations in at least thirty genes and still counting on. MCPH genes majorly code for a centrosomal protein subset, cell cycle proteins, nuclear proteins and cytoskeletal proteins. Centrosomal subset is the most highly characterized subset of MCPH proteins.Cep63andCdk5rap2with their paraloguesCcdc67andPde4dipplay important role for the normal assembly of spindles and in DNA damage response. They regulate neuroepithelial cell division in the developing brain, so decrease in centrosome number results in genomic instability. Study presented in this manuscript was aimed to identify the tissue specific expression of genes with their paralogues (Cep63andCcdc67) and (Cdk5rap2andPde4dip) in developing and adult normal mice tissue cells. For this purpose we successfully generated timed pregnant mice at the peak of neurogenesis and 36 samples were screened for the expression of these genes and their paralogues. RNA was extracted from the mice tissues using trizol reagent. Extracted RNA was reverse transcribed and cDNA was subjected to PCR amplification using gene specific primers, generated by using primer3 at the intron-exon junction to avoid the genomic interference. One house keeping gene β-actin was used as a positive control. This study reveals thatCep63 and Ccdc67paralogues are playing a suggestive collective role in the mice development as shown by multiple length bands.Cdk5rap2an MCPH gene and its paraloguePde4dipband suggested that maybe they might have redundant roles in body organs apart from the brain. These proteins have tissue-specific expression in all studied tissues from the developing and adult mice. We identified multiple bands in the different samples of all four genes which might reflect towards multiple transcript variants.