title: A human PKD1 transgene generates functional polycystin-1 in mice and is associated with a cystic phenotype.
abstract: Three founder transgenic mice were generated with a 108 kb human genomic fragment containing the entire autosomal dominant polycystic kidney disease (ADPKD) gene, PKD1, plus the tuberous sclerosis gene, TSC2. Two lines were established (TPK1 and TPK3) each with approximately 30 copies of the transgene. Both lines produced full-length PKD1 mRNA and polycystin-1 protein that was developmentally regulated, similar to the endogenous pattern, with expression during renal embryogenesis and neonatal life, markedly reduced at the conclusion of renal development. Tuberin expression was limited to the brain. Transgenic animals from both lines (and the TPK2 founder animal) often displayed a renal cystic phenotype, typically consisting of multiple microcysts, mainly of glomerular origin. Hepatic cysts and bile duct proliferation, characteristic of ADPKD, were also seen. All animals with two copies of the transgenic chromosome developed cysts and, in total, 48 of the 100 transgenic animals displayed a cystic phenotype. To test the functionality of the transgene, animals were bred with the Pkd1(del34) knockout mouse. Both transgenic lines rescued the embryonically lethal Pkd1(del34/del34) phenotype, demonstrating that human polycystin-1 can complement for loss of the endogenous protein. The rescued animals were viable into adulthood, although more than half developed hepatic cystic disease in later life, similar to the phenotype of older Pkd1(del34/+) animals. The TPK mice have defined a minimal area that appropriately expresses human PKD1. Furthermore, this model indicates that over-expression of normal PKD1 can elicit a disease phenotype, suggesting that the level of polycystin-1 expression may be relevant in the human disease.
authors.affiliation: Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK.
authors.forename: L
authors.initials: L
authors.lastname: Pritchard
authors.forename: J A
authors.initials: JA
authors.lastname: Sloane-Stanley
authors.forename: J A
authors.initials: JA
authors.lastname: Sharpe
authors.forename: R
authors.initials: R
authors.lastname: Aspinwall
authors.forename: W
authors.initials: W
authors.lastname: Lu
authors.forename: V
authors.initials: V
authors.lastname: Buckle
authors.forename: L
authors.initials: L
authors.lastname: Strmecki
authors.forename: D
authors.initials: D
authors.lastname: Walker
authors.forename: C J
authors.initials: CJ
authors.lastname: Ward
authors.forename: C E
authors.initials: CE
authors.lastname: Alpers
authors.forename: J
authors.initials: J
authors.lastname: Zhou
authors.forename: W G
authors.initials: WG
authors.lastname: Wood
authors.forename: P C
authors.initials: PC
authors.lastname: Harris
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D017353:Gene Deletion; D018628:Gene Dosage; D005816:Genetic Complementation Test; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000378:metabolism / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D023282:Nuclease Protection Assays; D010641:Phenotype; D016891:Polycystic Kidney, Autosomal Dominant / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012097:Repressor Proteins / Q000032:analysis; D050396:TRPP Cation Channels; D019076:Transgenes / Q000235:genetics*; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins
chemical_list: D011506:Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D050396:TRPP Cation Channels; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; C000624653:TSC2 protein, human; C000624654:Tsc2 protein, mouse