|
|
J Clin Invest. 1995 October; 96(4): 2051–2064. doi: 10.1172/JCI118253. | PMCID: PMC185844 |
A mouse model for the delta F508 allele of cystic fibrosis. B G Zeiher, E Eichwald, J Zabner, J J Smith, A P Puga, P B McCray, Jr, M R Capecchi, M J Welsh, and K R Thomas Howard Hughes Medical Institute, Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242, USA. Abstract The most common cause of cystic fibrosis is a mutation that deletes phenylalanine 508 in cystic fibrosis transmembrane conductance regulator (CFTR). The delta F508 protein is misprocessed and degraded rather than traveling to the apical membrane. We used a novel strategy to introduce the delta F508 mutation into the mouse CFTR gene. Affected epithelia from homozygous delta F508 mice lacked CFTR in the apical membrane and were Cl-impermeable. These abnormalities are the same as those observed in patients with delta F508 and suggest that these mice have the same cellular defect. 40% of homozygous delta F508 animals survived into adulthood and displayed several abnormalities found in human disease and in CFTR null mice. These animals should provide an excellent model to investigate pathogenesis and to examine therapies directed at correcting the delta F508 defect. Full text Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (6.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References. Images in this article Click on the image to see a larger version. These references are in PubMed. This may not be the complete list of references from this article. - Riordan JR, Rommens JM, Kerem B, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou JL, et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 1989 Sep 8;245(4922):1066–1073. [PubMed]
- Lemna WK, Feldman GL, Kerem B, Fernbach SD, Zevkovich EP, O'Brien WE, Riordan JR, Collins FS, Tsui LC, Beaudet AL. Mutation analysis for heterozygote detection and the prenatal diagnosis of cystic fibrosis. N Engl J Med. 1990 Feb 1;322(5):291–296. [PubMed]
- Cheng SH, Gregory RJ, Marshall J, Paul S, Souza DW, White GA, O'Riordan CR, Smith AE. Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis. Cell. 1990 Nov 16;63(4):827–834. [PubMed]
- Kartner N, Augustinas O, Jensen TJ, Naismith AL, Riordan JR. Mislocalization of delta F508 CFTR in cystic fibrosis sweat gland. Nat Genet. 1992 Aug;1(5):321–327. [PubMed]
- Engelhardt JF, Yankaskas JR, Ernst SA, Yang Y, Marino CR, Boucher RC, Cohn JA, Wilson JM. Submucosal glands are the predominant site of CFTR expression in the human bronchus. Nat Genet. 1992 Nov;2(3):240–248. [PubMed]
- Denning GM, Ostedgaard LS, Welsh MJ. Abnormal localization of cystic fibrosis transmembrane conductance regulator in primary cultures of cystic fibrosis airway epithelia. J Cell Biol. 1992 Aug;118(3):551–559. [PubMed]
- Knowles M, Gatzy J, Boucher R. Relative ion permeability of normal and cystic fibrosis nasal epithelium. J Clin Invest. 1983 May;71(5):1410–1417. [PubMed]
- Snouwaert JN, Brigman KK, Latour AM, Malouf NN, Boucher RC, Smithies O, Koller BH. An animal model for cystic fibrosis made by gene targeting. Science. 1992 Aug 21;257(5073):1083–1088. [PubMed]
- Ratcliff R, Evans MJ, Cuthbert AW, MacVinish LJ, Foster D, Anderson JR, Colledge WH. Production of a severe cystic fibrosis mutation in mice by gene targeting. Nat Genet. 1993 May;4(1):35–41. [PubMed]
- O'Neal WK, Hasty P, McCray PB, Jr, Casey B, Rivera-Pérez J, Welsh MJ, Beaudet AL, Bradley A. A severe phenotype in mice with a duplication of exon 3 in the cystic fibrosis locus. Hum Mol Genet. 1993 Oct;2(10):1561–1569. [PubMed]
- Dorin JR, Dickinson P, Alton EW, Smith SN, Geddes DM, Stevenson BJ, Kimber WL, Fleming S, Clarke AR, Hooper ML, et al. Cystic fibrosis in the mouse by targeted insertional mutagenesis. Nature. 1992 Sep 17;359(6392):211–215. [PubMed]
- Deng C, Thomas KR, Capecchi MR. Location of crossovers during gene targeting with insertion and replacement vectors. Mol Cell Biol. 1993 Apr;13(4):2134–2140. [PubMed]
- Tata F, Stanier P, Wicking C, Halford S, Kruyer H, Lench NJ, Scambler PJ, Hansen C, Braman JC, Williamson R, et al. Cloning the mouse homolog of the human cystic fibrosis transmembrane conductance regulator gene. Genomics. 1991 Jun;10(2):301–307. [PubMed]
- Yorifuji T, Lemna WK, Ballard CF, Rosenbloom CL, Rozmahel R, Plavsic N, Tsui LC, Beaudet AL. Molecular cloning and sequence analysis of the murine cDNA for the cystic fibrosis transmembrane conductance regulator. Genomics. 1991 Jul;10(3):547–550. [PubMed]
- Denning GM, Anderson MP, Amara JF, Marshall J, Smith AE, Welsh MJ. Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive. Nature. 1992 Aug 27;358(6389):761–764. [PubMed]
- Dalemans W, Barbry P, Champigny G, Jallat S, Dott K, Dreyer D, Crystal RG, Pavirani A, Lecocq JP, Lazdunski M. Altered chloride ion channel kinetics associated with the delta F508 cystic fibrosis mutation. Nature. 354(6354):526–528. [PubMed]
- Robertson E, Bradley A, Kuehn M, Evans M. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature. 1986 Oct 2;323(6087):445–448. [PubMed]
- Thomas KR, Capecchi MR. Targeted disruption of the murine int-1 proto-oncogene resulting in severe abnormalities in midbrain and cerebellar development. Nature. 1990 Aug 30;346(6287):847–850. [PubMed]
- Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987 Nov 6;51(3):503–512. [PubMed]
- Cathala G, Savouret JF, Mendez B, West BL, Karin M, Martial JA, Baxter JD. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. [PubMed]
- Tan SS, Weis JH. Development of a sensitive reverse transcriptase PCR assay, RT-RPCR, utilizing rapid cycle times. PCR Methods Appl. 1992 Nov;2(2):137–143. [PubMed]
- Mansour SL, Goddard JM, Capecchi MR. Mice homozygous for a targeted disruption of the proto-oncogene int-2 have developmental defects in the tail and inner ear. Development. 1993 Jan;117(1):13–28. [PubMed]
- Knowles MR, Clarke LL, Boucher RC. Activation by extracellular nucleotides of chloride secretion in the airway epithelia of patients with cystic fibrosis. N Engl J Med. 1991 Aug 22;325(8):533–538. [PubMed]
- Alton EW, Currie D, Logan-Sinclair R, Warner JO, Hodson ME, Geddes DM. Nasal potential difference: a clinical diagnostic test for cystic fibrosis. Eur Respir J. 1990 Sep;3(8):922–926. [PubMed]
- Githens S, Schexnayder JA, Moses RL, Denning GM, Smith JJ, Frazier ML. Mouse pancreatic acinar/ductular tissue gives rise to epithelial cultures that are morphologically, biochemically, and functionally indistinguishable from interlobular duct cell cultures. In Vitro Cell Dev Biol Anim. 1994 Sep;30A(9):622–635. [PubMed]
- Smith JJ, Welsh MJ. Fluid and electrolyte transport by cultured human airway epithelia. J Clin Invest. 1993 Apr;91(4):1590–1597. [PubMed]
- Ramírez-Solis R, Zheng H, Whiting J, Krumlauf R, Bradley A. Hoxb-4 (Hox-2.6) mutant mice show homeotic transformation of a cervical vertebra and defects in the closure of the sternal rudiments. Cell. 1993 Apr 23;73(2):279–294. [PubMed]
- Trezise AE, Buchwald M. In vivo cell-specific expression of the cystic fibrosis transmembrane conductance regulator. Nature. 1991 Oct 3;353(6343):434–437. [PubMed]
- Strong TV, Boehm K, Collins FS. Localization of cystic fibrosis transmembrane conductance regulator mRNA in the human gastrointestinal tract by in situ hybridization. J Clin Invest. 1994 Jan;93(1):347–354. [PubMed]
- Gaillard D, Ruocco S, Lallemand A, Dalemans W, Hinnrasky J, Puchelle E. Immunohistochemical localization of cystic fibrosis transmembrane conductance regulator in human fetal airway and digestive mucosa. Pediatr Res. 1994 Aug;36(2):137–143. [PubMed]
- Trezise AE, Linder CC, Grieger D, Thompson EW, Meunier H, Griswold MD, Buchwald M. CFTR expression is regulated during both the cycle of the seminiferous epithelium and the oestrous cycle of rodents. Nat Genet. 1993 Feb;3(2):157–164. [PubMed]
- Veeze HJ, Sinaasappel M, Bijman J, Bouquet J, de Jonge HR. Ion transport abnormalities in rectal suction biopsies from children with cystic fibrosis. Gastroenterology. 1991 Aug;101(2):398–403. [PubMed]
- Berschneider HM, Knowles MR, Azizkhan RG, Boucher RC, Tobey NA, Orlando RC, Powell DW. Altered intestinal chloride transport in cystic fibrosis. FASEB J. 1988 Jul;2(10):2625–2629. [PubMed]
- Goldstein JL, Shapiro AB, Rao MC, Layden TJ. In vivo evidence of altered chloride but not potassium secretion in cystic fibrosis rectal mucosa. Gastroenterology. 1991 Oct;101(4):1012–1019. [PubMed]
- Clarke LL, Grubb BR, Gabriel SE, Smithies O, Koller BH, Boucher RC. Defective epithelial chloride transport in a gene-targeted mouse model of cystic fibrosis. Science. 1992 Aug 21;257(5073):1125–1128. [PubMed]
- Marino CR, Matovcik LM, Gorelick FS, Cohn JA. Localization of the cystic fibrosis transmembrane conductance regulator in pancreas. J Clin Invest. 1991 Aug;88(2):712–716. [PubMed]
- McCarty NA, McDonough S, Cohen BN, Riordan JR, Davidson N, Lester HA. Voltage-dependent block of the cystic fibrosis transmembrane conductance regulator Cl- channel by two closely related arylaminobenzoates. J Gen Physiol. 1993 Jul;102(1):1–23. [PubMed]
- Lukacs GL, Chang XB, Bear C, Kartner N, Mohamed A, Riordan JR, Grinstein S. The delta F508 mutation decreases the stability of cystic fibrosis transmembrane conductance regulator in the plasma membrane. Determination of functional half-lives on transfected cells. J Biol Chem. 1993 Oct 15;268(29):21592–21598. [PubMed]
- Ward CL, Kopito RR. Intracellular turnover of cystic fibrosis transmembrane conductance regulator. Inefficient processing and rapid degradation of wild-type and mutant proteins. J Biol Chem. 1994 Oct 14;269(41):25710–25718. [PubMed]
- Puchelle E, Gaillard D, Ploton D, Hinnrasky J, Fuchey C, Boutterin MC, Jacquot J, Dreyer D, Pavirani A, Dalemans W. Differential localization of the cystic fibrosis transmembrane conductance regulator in normal and cystic fibrosis airway epithelium. Am J Respir Cell Mol Biol. 1992 Nov;7(5):485–491. [PubMed]
- Trapnell BC, Chu CS, Paakko PK, Banks TC, Yoshimura K, Ferrans VJ, Chernick MS, Crystal RG. Expression of the cystic fibrosis transmembrane conductance regulator gene in the respiratory tract of normal individuals and individuals with cystic fibrosis. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6565–6569. [PubMed]
- Boucher RC, Stutts MJ, Knowles MR, Cantley L, Gatzy JT. Na+ transport in cystic fibrosis respiratory epithelia. Abnormal basal rate and response to adenylate cyclase activation. J Clin Invest. 1986 Nov;78(5):1245–1252. [PubMed]
- Behm JK, Hagiwara G, Lewiston NJ, Quinton PM, Wine JJ. Hyposecretion of beta-adrenergically induced sweating in cystic fibrosis heterozygotes. Pediatr Res. 1987 Sep;22(3):271–276. [PubMed]
- Sato K, Sato F. Variable reduction in beta-adrenergic sweat secretion in cystic fibrosis heterozygotes. J Lab Clin Med. 1988 May;111(5):511–518. [PubMed]
- Gabriel SE, Brigman KN, Koller BH, Boucher RC, Stutts MJ. Cystic fibrosis heterozygote resistance to cholera toxin in the cystic fibrosis mouse model. Science. 1994 Oct 7;266(5182):107–109. [PubMed]
- Cuthbert AW, Halstead J, Ratcliff R, Colledge WH, Evans MJ. The genetic advantage hypothesis in cystic fibrosis heterozygotes: a murine study. J Physiol. 1995 Jan 15;482 (:449–454. [PubMed]
|
This article has been 
