Transgenic mouse as a model of lipoprotein glomerulopathy (LPG), and analysis of LPG

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Lipoprotein glomerulopathy (LPG) is a newly recognized renal disease in 1988. It is characterized by abnormal lipoprotein deposition in the marked dilated glomerular capillaries, and elevated plasma concentration of apolipoprotein (apo) E.Recently, we identified an apoE variant, apoE2 Sendai (Arg 145Pro) in three Japanese kindred with LPG.Furthermore, we discovered three different novel apoE variants, apo E2 Kyoto (Arg25Cys) apo E1 (del 141-143) and apo E1 (del 156-173), which are closely related to LPG.Establishment of an apo E 1 (del 141-143) transgenic mouse was tried as an animal model of LPG.ApoE1 (del 141-143) variant was introduced into normal apoE gene including promoter region (11.5kb) using site directed mutagenesis. Fragment of the variant human apoE gene digested with HindIII was purified and microinjected into pronuclei of fertilized eggs of C57BL/6 mice. Injected eggs were surgically transferred to oviducts of anesthetized surrogate females. However, the transgenic mice were killed because they were infected with pathogenic virus. Second microinjection was done, and seven pups were born. Genomic DNA was isolated from their tails. Polymerase chain reaction analysis was performed on genomic tail DNA using primer set of human apoE gene. Human apoE gene was detected from a female mouse. Human apoE gene detection rate of Fl mice was very low, and human apoE gene was confirmed currently in two male mice. After homozygotes of apoE1 (del 141-143) mice are established, analysis of their glomerulus lesions and serum lipoproteins are planned.