Gene mapping of glaucoma related phenotypes in a forward genetics ENU screen.

Document Type


Publication Date

Summer 2016

JAX Location

In: Student Reports, Summer 2016, Jackson Laboratory


Glaucoma is characterized by the loss of retinal ganglion cells and optic nerve degeneration; the genetic causes are poorly understood. Our hypothesis is that genetic mapping can be used to identify candidate mutations related to a specific glaucoma phenotype of the strain ENU.D0139, and careful study of these mutations will allow us to identify the causative mutation and disease mechanism. Whole Genome Sequencing data from a pool of ENU.D0139 was analyzed to determine and evaluate if the mutations identified were real. PCR was performed and the product was sent for purification and Sanger Sequencing in order to perform fine mapping. Previous data showed that chromosomes 4 and 14 are gregions of interest, therefore thirty-five mutants were mapped and chromosomes sequenced for mutations in those chromosomes. Chromosome 14 was eliminated as a candidate region, because some mice were wildtype throughout. On the other hand, 33 mutants were heterozygous throughout the chromosome 4 and two were recombinant within the region. The region of interest was narrowed down to approximately 3Mb containing mutations in four candidate genes. My study focused on the strongest candidate gene: Macf1. It is involved in cell polarization, maintenance of tissue integrity, and cellular movements. A western blot was done and no differences were seen between wildtype and mutant mice for MACF1. Immunohistochemistry showed that MACF1 may be present in the ciliary body. Further experiments should be conducted determine which isoforms of the protein the antibody binds.

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