The Generation Of CysLT1Gene Knockout Mice And H99-GFP Strain And The Study Of CysLT1Receptor Distribution In Cerebral Microvessel And Hippocampus Of Mice

Aim:1. to generate CysLT1R-deficient mice and H99-GFP strain.2. to study the expression of CysLT1receptor in cerebral microvasculature andhippocampus of mice infected with Cryptococcus neoformans.H99.Methods:1. CysLT1gene-disrupted mice were detected by PCR, RT-PCR and Western-blot.2. GFP plasmids were integrated into the genome of the H99by a BioRad PDS-1000/Hebiolistic particle delivery system.3. Mice model of experimental meningitis was prepared by intratracheal inoculation ofC.neoformans.H99.4. The expression of CysLT1receptor in brain tissues were detected by immunohistoche-mistry and immunofluorescence technique.Results:1. CysLT1gene-disrupted mice we have generated showed no expression of CysLT1receptor at both nucleic acid and protein levers.2. GFP plasmids were integrated into the genome of the H99by biolistic particle deliverysystem, generating a novel H99strain which stably express high level of GFP.3. Compared with sham group, wild-type mice infected with Cryptococcus neoformansshowed higher level expression of CysLT1receptor in the neuron of cortex andhippocampus, especially in the endothelial cell of cerebral microvessel.Conclusion:1. This study has successfully generated the CysLT1gene knockout mice and H99-GFPstrain, which may be the powerful tools for the research projects such as the role ofCysLT1receptor in Cryptococcus invasion of the cerebral microvessel.2. For the first time we have found the higher level expression of CysLT1receptor in theendothelial cell of cerebral microvessel as well as the neuron of cortex and hippocampusafter mice infected with Cryptococcus neoformans, which may have an importantcontribution to the future research projects in our laboratory.