JCB logo
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 1532K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kielty, C. M.
Right arrow Articles by Jimenez, S. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kielty, C. M.
Right arrow Articles by Jimenez, S. A.
Right arrowPubmed/NCBI databases
*Gene*GEO Profiles
*HomoloGene*OMIM
*UniGene
*Compound via MeSH
*Substance via MeSH
Hazardous Substances DB
*ETHYLENEDIAMINE TETRAACETIC ACID
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/1998//1159 $5.00
The Journal of Cell Biology, Volume 140, Number 5, , 1998 1159-1166

Article

The Tight Skin Mouse: Demonstration of Mutant Fibrillin-1 Production and Assembly into Abnormal Microfibrils



Cay M. Kielty*, Michael Raghunath{ddagger}, Linda D. Siracusa§, Michael J. Sherratt*, Reiner Peters||, C. Adrian Shuttleworth*, and Sergio A. Jimenez§

* School of Biological Sciences, University of Manchester, Manchester, M13 9PT, United Kingdom; {ddagger} Department of Dermatology, || Department of Medical Physics and Biophysics, University of Muenster, D-48149, Germany; and Department of Microbiology and Immunology, § Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-5541

Mice carrying the Tight skin (Tsk) mutation harbor a genomic duplication within the fibrillin-1 (Fbn 1) gene that results in a larger than normal in-frame Fbn 1 transcript. In this study, the consequences of the Tsk mutation for fibrillin-containing microfibrils have been examined. Dermal fibroblasts from Tsk/+ mice synthesized and secreted both normal fibrillin (~330 kD) and the mutant oversized Tsk fibrillin-1 (~450 kD) in comparable amounts, and Tsk fibrillin-1 was stably incorporated into cell layers. Immunohistochemical and ultrastructural analyses of normal and Tsk/+ mouse skin highlighted differences in the gross organization and distribution of microfibrillar arrays. Rotary shadowing of high Mr preparations from Tsk/+ skin demonstrated the presence of abundant beaded microfibrils. Some of these had normal morphology and periodicity, but others were distinguished by diffuse interbeads, longer periodicity, and tendency to aggregate. The presence of a structurally abnormal population of microfibrils in Tsk/+ skin was unequivocally demonstrated after calcium chelation and in denaturating conditions. Scanning transmission electron microscopy highlighted the presence of more mass in Tsk/+ skin microfibrils than in normal mice skin microfibrils. These data indicate that Tsk fibrillin-1 polymerizes and becomes incorporated into a discrete population of beaded microfibrils with altered molecular organization.

Abbreviations used in this paper: cbEGF, calcium-binding EGF; cys, cysteine; GuHCL, guanidinium chloride; Fbn 1, fibrillin 1 gene; met, methionine; MFS, Marfan syndrome; STEM, scanning transmission electron microscopy; Tsk, Tight skin mutation.

C.M. Kielty thanks S. Whittaker (University of Manchester), and S.A. Jimenez thanks R. McGrath for expert technical assistance. M. Raghunath thanks B. Nusgens (University of Leige, Leige, Belgium), for skin specimens of normal and Tsk/+ mice, and M. Tschodrich-Rotter (University of Muenster) for help with the confocal laser scanning studies.

This research was supported in part by the Medical Research Council, UK (grant G9227295MA), the U.S. National Institute of Health (grant ARO-32564) to S.A. Jimenez., European Molecular Biology Organization short-term fellowship to M. Raghunath (grant ASTF 8516), and the Deutsche Forschung-Gemeinschaft (grants Ra447/3-1 and Ra 447/3-2).

Address all correspondence to Cay M. Kielty, School of Biological Sciences, 2.205 Stopford Bldg., University of Manchester, Manchester M13 9PT, UK. Tel.: (44) 161-275-5739. Fax: (44) 161-275-5082. E-mail: cay. kielty{at}man.ac.uk


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents