Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Jul 1;118(1):203–212. doi: 10.1083/jcb.118.1.203

Chondrodysplasia in transgenic mice harboring a 15-amino acid deletion in the triple helical domain of pro alpha 1(II) collagen chain

PMCID: PMC2289514  PMID: 1618904

Abstract

We have generated transgenic mice by microinjection of a 39-kb mouse pro alpha 1(II) collagen gene construct containing a deletion of exon 7 and intron 7. This mutation was expected to disturb the assembly and processing of the homotrimeric type II collagen molecule in cartilage. Expression of transgene mRNA at levels equivalent or higher than the endogenous mRNA in the offspring of two founder animals resulted in a severe chondrodysplastic phenotype with short limbs, hypoplastic thorax, abnormal craniofacial development, and other skeletal deformities. The affected pups died at birth due to respiratory distress. Light microscopy of epiphyseal growth plates of transgenic pups demonstrated a marked reduction in cartilaginous extracellular matrix and disruption of the normal organization of the growth plate. The zone of proliferating chondrocytes was greatly reduced whereas the zone of hypertrophic chondrocytes was markedly increased extending deep into the diaphysis suggestive of a defect in endochondral ossification. Electron microscopic examination revealed chondrocytes with extended RER, a very severe reduction in the amount of cartilage collagen fibrils, and abnormalities in their structure. We postulate that the deletion in the alpha 1(II) collagen acts as a dominant negative mutation disrupting the assembly and secretion of type II collagen molecules. The consequences of the mutation include interference with normal endochondral ossification. These mice constitute a valuable model to study the mechanisms underlying human chondrodysplasias and normal bone formation.

Full Text

The Full Text of this article is available as a PDF (5.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ahmad N. N., Ala-Kokko L., Knowlton R. G., Jimenez S. A., Weaver E. J., Maguire J. I., Tasman W., Prockop D. J. Stop codon in the procollagen II gene (COL2A1) in a family with the Stickler syndrome (arthro-ophthalmopathy). Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6624–6627. doi: 10.1073/pnas.88.15.6624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ala-Kokko L., Baldwin C. T., Moskowitz R. W., Prockop D. J. Single base mutation in the type II procollagen gene (COL2A1) as a cause of primary osteoarthritis associated with a mild chondrodysplasia. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6565–6568. doi: 10.1073/pnas.87.17.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Byers P. H. Inherited disorders of collagen gene structure and expression. Am J Med Genet. 1989 Sep;34(1):72–80. doi: 10.1002/ajmg.1320340114. [DOI] [PubMed] [Google Scholar]
  4. Cheah K. S., Lau E. T., Au P. K., Tam P. P. Expression of the mouse alpha 1(II) collagen gene is not restricted to cartilage during development. Development. 1991 Apr;111(4):945–953. doi: 10.1242/dev.111.4.945. [DOI] [PubMed] [Google Scholar]
  5. Chelly J., Concordet J. P., Kaplan J. C., Kahn A. Illegitimate transcription: transcription of any gene in any cell type. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2617–2621. doi: 10.1073/pnas.86.8.2617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Dombrowski K. E., Vogel B. E., Prockop D. J. Mutations that alter the primary structure of type I procollagen have long-range effects on its cleavage by procollagen N-proteinase. Biochemistry. 1989 Aug 22;28(17):7107–7112. doi: 10.1021/bi00443a048. [DOI] [PubMed] [Google Scholar]
  8. Francomano C. A., Liberfarb R. M., Hirose T., Maumenee I. H., Streeten E. A., Meyers D. A., Pyeritz R. E. The Stickler syndrome: evidence for close linkage to the structural gene for type II collagen. Genomics. 1987 Dec;1(4):293–296. doi: 10.1016/0888-7543(87)90027-9. [DOI] [PubMed] [Google Scholar]
  9. Garofalo S., Vuorio E., Metsaranta M., Rosati R., Toman D., Vaughan J., Lozano G., Mayne R., Ellard J., Horton W. Reduced amounts of cartilage collagen fibrils and growth plate anomalies in transgenic mice harboring a glycine-to-cysteine mutation in the mouse type II procollagen alpha 1-chain gene. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9648–9652. doi: 10.1073/pnas.88.21.9648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horton W., Miyashita T., Kohno K., Hassell J. R., Yamada Y. Identification of a phenotype-specific enhancer in the first intron of the rat collagen II gene. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8864–8868. doi: 10.1073/pnas.84.24.8864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Knowlton R. G., Katzenstein P. L., Moskowitz R. W., Weaver E. J., Malemud C. J., Pathria M. N., Jimenez S. A., Prockop D. J. Genetic linkage of a polymorphism in the type II procollagen gene (COL2A1) to primary osteoarthritis associated with mild chondrodysplasia. N Engl J Med. 1990 Feb 22;322(8):526–530. doi: 10.1056/NEJM199002223220807. [DOI] [PubMed] [Google Scholar]
  12. Lee B., Vissing H., Ramirez F., Rogers D., Rimoin D. Identification of the molecular defect in a family with spondyloepiphyseal dysplasia. Science. 1989 May 26;244(4907):978–980. doi: 10.1126/science.2543071. [DOI] [PubMed] [Google Scholar]
  13. Maroteaux P., Stanescu V., Stanescu R., Le Marec B., Moraine C., Lejarraga H. Opsismodysplasia: a new type of chondrodysplasia with predominant involvement of the bones of the hand and the vertebrae. Am J Med Genet. 1984 Sep;19(1):171–182. doi: 10.1002/ajmg.1320190117. [DOI] [PubMed] [Google Scholar]
  14. McLeod M. J. Differential staining of cartilage and bone in whole mouse fetuses by alcian blue and alizarin red S. Teratology. 1980 Dec;22(3):299–301. doi: 10.1002/tera.1420220306. [DOI] [PubMed] [Google Scholar]
  15. Mendler M., Eich-Bender S. G., Vaughan L., Winterhalter K. H., Bruckner P. Cartilage contains mixed fibrils of collagen types II, IX, and XI. J Cell Biol. 1989 Jan;108(1):191–197. doi: 10.1083/jcb.108.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Metsäranta M., Toman D., de Crombrugghe B., Vuorio E. Mouse type II collagen gene. Complete nucleotide sequence, exon structure, and alternative splicing. J Biol Chem. 1991 Sep 5;266(25):16862–16869. [PubMed] [Google Scholar]
  17. Metsäranta M., Young M. F., Sandberg M., Termine J., Vuorio E. Localization of osteonectin expression in human fetal skeletal tissues by in situ hybridization. Calcif Tissue Int. 1989 Sep;45(3):146–152. doi: 10.1007/BF02556057. [DOI] [PubMed] [Google Scholar]
  18. Palotie A., Väisänen P., Ott J., Ryhänen L., Elima K., Vikkula M., Cheah K., Vuorio E., Peltonen L. Predisposition to familial osteoarthrosis linked to type II collagen gene. Lancet. 1989 Apr 29;1(8644):924–927. doi: 10.1016/s0140-6736(89)92507-5. [DOI] [PubMed] [Google Scholar]
  19. Poole A. R., Pidoux I., Reiner A., Choi H., Rosenberg L. C. Association of an extracellular protein (chondrocalcin) with the calcification of cartilage in endochondral bone formation. J Cell Biol. 1984 Jan;98(1):54–65. doi: 10.1083/jcb.98.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Prockop D. J., Baldwin C. T., Constantinou C. D. Mutations in type I procollagen genes that cause osteogenesis imperfecta. Adv Hum Genet. 1990;19:105–132. doi: 10.1007/978-1-4757-9065-8_2. [DOI] [PubMed] [Google Scholar]
  21. Sandberg M., Vuorio E. Localization of types I, II, and III collagen mRNAs in developing human skeletal tissues by in situ hybridization. J Cell Biol. 1987 Apr;104(4):1077–1084. doi: 10.1083/jcb.104.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Savagner P., Miyashita T., Yamada Y. Two silencers regulate the tissue-specific expression of the collagen II gene. J Biol Chem. 1990 Apr 25;265(12):6669–6674. [PubMed] [Google Scholar]
  23. Spranger J., Maroteaux P. The lethal osteochondrodysplasias. Adv Hum Genet. 1990;19:1-103, 331-2. doi: 10.1007/978-1-4757-9065-8_1. [DOI] [PubMed] [Google Scholar]
  24. Thorogood P., Bee J., von der Mark K. Transient expression of collagen type II at epitheliomesenchymal interfaces during morphogenesis of the cartilaginous neurocranium. Dev Biol. 1986 Aug;116(2):497–509. doi: 10.1016/0012-1606(86)90150-8. [DOI] [PubMed] [Google Scholar]
  25. Tiller G. E., Rimoin D. L., Murray L. W., Cohn D. H. Tandem duplication within a type II collagen gene (COL2A1) exon in an individual with spondyloepiphyseal dysplasia. Proc Natl Acad Sci U S A. 1990 May;87(10):3889–3893. doi: 10.1073/pnas.87.10.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vandenberg P., Khillan J. S., Prockop D. J., Helminen H., Kontusaari S., Ala-Kokko L. Expression of a partially deleted gene of human type II procollagen (COL2A1) in transgenic mice produces a chondrodysplasia. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7640–7644. doi: 10.1073/pnas.88.17.7640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vissing H., D'Alessio M., Lee B., Ramirez F., Godfrey M., Hollister D. W. Glycine to serine substitution in the triple helical domain of pro-alpha 1 (II) collagen results in a lethal perinatal form of short-limbed dwarfism. J Biol Chem. 1989 Nov 5;264(31):18265–18267. [PubMed] [Google Scholar]
  28. Vuorio E., de Crombrugghe B. The family of collagen genes. Annu Rev Biochem. 1990;59:837–872. doi: 10.1146/annurev.bi.59.070190.004201. [DOI] [PubMed] [Google Scholar]
  29. Wood A., Ashhurst D. E., Corbett A., Thorogood P. The transient expression of type II collagen at tissue interfaces during mammalian craniofacial development. Development. 1991 Apr;111(4):955–968. doi: 10.1242/dev.111.4.955. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES