Advertisement

Human Genetics

, Volume 65, Issue 1, pp 76–78 | Cite as

Manifestation of the fragile site Xq27 in fibroblasts

III. A method to demonstrate R-type replication patterns and the fragile site
  • G. Barbi
  • P. Steinbach
  • A. Wiedenmann
  • W. Vogel
Short Communications

Summary

A protocol is reported which allows the efficient induction of bromodeoxyridine (BrdU)-induced R-type replication patterns in fibroblast cultures prepared to demonstrate the fragile site fra(X)(q27). The technique includes partial synchronization of the culture by fluorodeoxyuridine (FdU) blocking at the G1/S transition. This block does not impair the induction of the fragile site in medium 199 containing methotrexate. The marked increase of the mitotic index in the synchronized culture may be an advantage in the study of folic acid sensitive fragile sites in fibroblasts. Adding BrdU after block removal leads to an efficient labeling of replicating chromosomes without severely impairing the manifestation of fra(X)(q27).

Keywords

Internal Medicine Methotrexate Folic Acid Metabolic Disease Mitotic Index 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Albertini RJ, DeMars (1974) Mosaicism of peripheral blood lymphocyte populations in females heterozygous for the Lesch-Nyhan mutation. Biochem Genet 11:397–411Google Scholar
  2. Barbi G, Steinbach P (1982) Increase in the incidence of the fragile site Xq27 in prometaphases. Hum Genet 61:82Google Scholar
  3. Chudley AE, Knoll J, Gerrard JW, Shepel L, McGahey E, Anderson J (1983) Fragile (X) X-linked mental retation. I: Relationship between age and intelligence and the frequency of expression of fragile(X)(q28). Am J Med Genet 14:699–712Google Scholar
  4. Epplen JT, Siebers JW, Vogel W (1975) DNA replication patterns of human chromosomes from fibroblasts and amniotic fluid cells revealed by a Giemsa staining method. Cytogenet Cell Genet 15: 177–185Google Scholar
  5. Froster-Iskenius U, Schwinger E, Weigert M, Fonatsch C (1982) Replication pattern in XXY cells with fra(X). Hum Genet 60: 278–280Google Scholar
  6. Howell RT, McDermott A (1982) Replication status of the fragile X chromosome, fra(X)(q27), in three heterozygous females. Hum Genet 62:282–284Google Scholar
  7. Lubs HA (1969) A marker X chromosome. Am J Hum Genet 21: 231–244Google Scholar
  8. McDonald JA, Kelley WN (1972) Lesch-Nyhan syndrome: Absence of the mutant enzyme in erythrocytes of a heterozygote for both normal and mutant hypoxanthine-guanine phosphoribosyl transferase. Biochem Genet 6:21–26Google Scholar
  9. Steinbach P, Barbi G, Baur S, Wiedenmann A (1983a) Expression of the fragile site Xq27 in fibroblasts. I. Detection of fra(X)(q27) in fibroblast clones from males with X-linked mental retardation. Hum Genet 63:404–405Google Scholar
  10. Steinbach P, Barbi G, Baur S, Vogel W (1983b) Expression of the fragile site Xq27 in fibroblasts. II. Detection of fra(X)-negative and fra(X)-positive clones from female carriers for the Martin-Bell syndrome. Hum Genet 64:279–282Google Scholar
  11. Sutherland GR (1979) Heritable fragile sites on human chromosomes. I. Factors affecting expression in lymphocyte culture. Am J Hum Genet 31:125–135Google Scholar
  12. Uchida IA, Joyce EM (1982) Activity of the fragile X in heterozygous carriers. Am J Hum Genet 34:286–293Google Scholar
  13. Vogel W, Schempp W, Sigwarth I (1978) Comparison of thymidine, fluorodeoxyridine, hydroxyurea, and methotrexate blocking at the G1/S phase transition of the cell cycle, studied by replication patterns. Hum Genet 45:183–198Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • G. Barbi
    • 1
  • P. Steinbach
    • 1
  • A. Wiedenmann
    • 1
  • W. Vogel
    • 1
  1. 1.Abteilung Klinische GenetikUniversität UlmUlm/DonauFederal Republic of Germany

Personalised recommendations