Cell motility and microtubules in cultured fibroblasts from patients with Kartagener syndrome.

Patients with Kartagener syndrome (KS) show defects in ciliary and flagellar movement that are usually associated with the partial or total absence of dynein side arms from axonemal microtubules. Dynein is essential for such movements, but its involvement in other cellular (particularly microtubule-related) processes is unknown. It has recently been reported that neutrophils from KS patients show impaired motility including responses to chemotactic stimuli, suggesting that dynein-like proteins may be generally involved in motile processes. In support of this, we have now found that spontaneous motility of cultured skin fibroblasts from KS patients is also markedly impaired. Three cell lines derived from skin explants of KS patients with deficient dynein side arms in nasal cilia and eight cell lines derived from normal volunteers were studied. Fibroblasts were seeded into dishes containing colloidal gold-coated cover glasses [Albrecht-Buehler, 1977], incubated for 24h at 37 degrees C, and the area of cell "phagokinetic" tracks determined. Each cell line studied in this manner reproducibly displayed an amount of spontaneous motility characteristic for that cell line. The mean track area (+/- SE) for all control cells studied was 14.6 +/- 0.5 X 10(3) micron2 whereas for KS fibroblasts was 8.7 +/- 0.4 X 10(3) micron2 (P less than 0.001). Immunofluorescence microscopy using antitubulin and antihuman 210 K MAP antibodies revealed no differences in the staining patterns between control and KS fibroblasts. Pinocytic rates were identical, and the complement of tubulin and major microtubule associated proteins as seen on one-dimensional SDS polyacrylamide gel autoradiographs appeared similar for control and KS cells. Thus, the observed motility defect is probably not the result of alterations in the occurrence or distribution of microtubules or in the occurrence or binding of the major microtubule-associated proteins. This defect in cellular motility may be related to the absence of dynein or may reflect another independent cellular defect.

[1]  J. Palmblad,et al.  Structure and function of neutrophil leukocytes from patients with the immotile-cilia syndrome. , 2009, Acta medica Scandinavica.

[2]  H. Malech,et al.  Abnormal movement of polymorphonuclear neutrophils in the Immotile Cilia Syndrome. Cinemicrographic analysis. , 1981, Experimental cell research.

[3]  G. Borisy,et al.  Immunofluorescence localization of HeLa cell microtubule-associated proteins on microtubules in vitro and in vivo , 1980, The Journal of cell biology.

[4]  J. Oliver,et al.  Polarization of endocytosis and receptor topography on cultured macrophages , 1980, The Journal of cell biology.

[5]  J. Rosenbaum,et al.  Dynein binds to and crossbridges cytoplasmic microtubules. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[6]  W. Sale,et al.  Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella , 1979, The Journal of cell biology.

[7]  G. Albrecht-Buehler,et al.  The orientation of centrioles in migrating 3T3 cells. , 1979, Experimental cell research.

[8]  S. M. Wolniak,et al.  Chromosome movement in lysed mitotic cells is inhibited by vanadate , 1978, The Journal of cell biology.

[9]  J. Oliver,et al.  Analogous ultrastructure and surface properties during capping and phagocytosis in leukocytes , 1978, The Journal of cell biology.

[10]  K. Weber,et al.  The display of microtubules in transformed cells , 1977, Cell.

[11]  G. Albrecht-Buehler,et al.  Phagokinetic tracks of 3T3 cells: Parallels between the orientation of track segments and of cellular structures which contain actin or tubulin , 1977, Cell.

[12]  B. Afzelius.  A human syndrome caused by immotile cilia. , 1976, Science.

[13]  T. Pollard,et al.  A dynein‐like protein from brain , 1974, FEBS letters.

[14]  C. Cantor,et al.  A dynein‐like protein associated with neurotubules , 1974, FEBS letters.

[15]  C. Cantor,et al.  Microtubule assembly in the absence of added nucleotides. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[16]  I. Gibbons,et al.  FLAGELLAR MOVEMENT AND ADENOSINE TRIPHOSPHATASE ACTIVITY IN SEA URCHIN SPERM EXTRACTED WITH TRITON X-100 , 1972, The Journal of cell biology.

[17]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[18]  P. Satir.  STUDIES ON CILIA , 1968, The Journal of cell biology.

[19]  L. Holmes,et al.  A Reappraisal of Kartagener's Syndrome , 1968, The American journal of the medical sciences.

[20]  A. Rowe,et al.  Dynein: A Protein with Adenosine Triphosphatase Activity from Cilia , 1965, Science.

[21]  J. Rosenbaum,et al.  Dynein binding to microtubules containing microtubule-associated proteins. , 1981, Cell motility.

[22]  P. Wilkinson,et al.  A visual analysis of chemotactic and chemokinetic locomotion of human neutrophil leucocytes. Use of a new chemotaxis assay with Candida albicans as gradient source. , 1978, Experimental cell research.

[23]  H. Sakai,et al.  INDUCTION OF CHROMOSOME MOTION IN THE GLYCEROL‐ISOLATED MITOTIC APPARATUS: NUCLEOTIDE SPECIFICITY AND EFFECTS OF ANTIDYNEIN AND MYOSIN SERA ON THE MOTION * , 1976, Development, growth & differentiation.

[24]  A. Allison.  Mechanism of movement and maintenance of polarity in leucocytes. , 1974, Antibiotics and chemotherapy.

[25]  L. Rebhun.  Polarized intracellular particle transport: saltatory movements and cytoplasmic streaming. , 1972, International review of cytology.