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. 1994 Nov;55(5):951–959.

Molecular Analysis and Test of Linkage between the FMR-1 Gene and Infantile Autism in Multiplex Families

Joachim Hallmayer, Elizabeth Pintado, Linda Lotspeich, Donna Spiker, William McMahon, P Brent Petersen, Peter Nicholas, Carmen Pingree, Helena C Kraemer, Dona Lee Wong, Edward Ritvo, Alice Lin, Joan Hebert, Luigi L Cavalli-Sforza, Roland D Ciaranello
PMCID: PMC1918316  PMID: 7977358

Abstract

Approximately 2%–5% of autistic children show cytogenetic evidence of the fragile X syndrome. This report tests whether infantile autism in multiplex autism families arises from an unusual manifestion of the fragile X syndrome. This could arise either by expansion of the (CGG)n trinucleotide repeat in FMR-1 or from a mutation elsewhere in the gene. We studied 35 families that met stringent criteria for multiplex autism. Amplification of the trinucleotide repeat and analysis of methylation status were performed in 79 autistic children and in 31 of their unaffected siblings, by Southern blot analysis. No examples of amplified repeats were seen in the autistic or control children or in their parents or grandparents. We next examined the hypothesis that there was a mutation elsewhere in the FMR-1 gene, by linkage analysis in 32 of these families. We tested four different dominant models and a recessive model. Linkage to FMR-1 could be excluded (lod score between −24 and −62) in all models by using probes DXS548, FRAXAC1, and FRAXAC2 and the CGG repeat itself. Tests for heterogeneity in this sample were negative, and the occurrence of positive lod scores in this data set could be attributed to chance. Analysis of the data by the affected-sib method also did not show evidence for linkage of any marker to autism. These results enable us to reject the hypothesis that multiplex autism arises from expansion of the (CGG)n trinucleotide repeat in FMR-1. Further, because the overall lod scores for all probes in all models tested were highly negative, linkage to FMR-1 can also be ruled out in multiplex autistic families.

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Selected References

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

  1. Anderson M. A., Gusella J. F. Use of cyclosporin A in establishing Epstein-Barr virus-transformed human lymphoblastoid cell lines. In Vitro. 1984 Nov;20(11):856–858. doi: 10.1007/BF02619631. [DOI] [PubMed] [Google Scholar]
  2. De Boulle K., Verkerk A. J., Reyniers E., Vits L., Hendrickx J., Van Roy B., Van den Bos F., de Graaff E., Oostra B. A., Willems P. J. A point mutation in the FMR-1 gene associated with fragile X mental retardation. Nat Genet. 1993 Jan;3(1):31–35. doi: 10.1038/ng0193-31. [DOI] [PubMed] [Google Scholar]
  3. Einfeld S., Hall W. Behavior phenotype of the fragile X syndrome. 1992 Apr 15-May 1Am J Med Genet. 43(1-2):56–60. doi: 10.1002/ajmg.1320430108. [DOI] [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Fisch G. S. Is autism associated with the fragile X syndrome? 1992 Apr 15-May 1Am J Med Genet. 43(1-2):47–55. doi: 10.1002/ajmg.1320430107. [DOI] [PubMed] [Google Scholar]
  6. Folstein S. E., Piven J. Etiology of autism: genetic influences. Pediatrics. 1991 May;87(5 Pt 2):767–773. [PubMed] [Google Scholar]
  7. Gedeon A. K., Baker E., Robinson H., Partington M. W., Gross B., Manca A., Korn B., Poustka A., Yu S., Sutherland G. R. Fragile X syndrome without CCG amplification has an FMR1 deletion. Nat Genet. 1992 Aug;1(5):341–344. doi: 10.1038/ng0892-341. [DOI] [PubMed] [Google Scholar]
  8. Jorde L. B., Hasstedt S. J., Ritvo E. R., Mason-Brothers A., Freeman B. J., Pingree C., McMahon W. M., Petersen B., Jenson W. R., Mo A. Complex segregation analysis of autism. Am J Hum Genet. 1991 Nov;49(5):932–938. [PMC free article] [PubMed] [Google Scholar]
  9. Knight S. J., Flannery A. V., Hirst M. C., Campbell L., Christodoulou Z., Phelps S. R., Pointon J., Middleton-Price H. R., Barnicoat A., Pembrey M. E. Trinucleotide repeat amplification and hypermethylation of a CpG island in FRAXE mental retardation. Cell. 1993 Jul 16;74(1):127–134. doi: 10.1016/0092-8674(93)90300-f. [DOI] [PubMed] [Google Scholar]
  10. Lathrop G. M., Lalouel J. M. Easy calculations of lod scores and genetic risks on small computers. Am J Hum Genet. 1984 Mar;36(2):460–465. [PMC free article] [PubMed] [Google Scholar]
  11. Le Couteur A., Rutter M., Lord C., Rios P., Robertson S., Holdgrafer M., McLennan J. Autism diagnostic interview: a standardized investigator-based instrument. J Autism Dev Disord. 1989 Sep;19(3):363–387. doi: 10.1007/BF02212936. [DOI] [PubMed] [Google Scholar]
  12. Lord C., Rutter M., Goode S., Heemsbergen J., Jordan H., Mawhood L., Schopler E. Autism diagnostic observation schedule: a standardized observation of communicative and social behavior. J Autism Dev Disord. 1989 Jun;19(2):185–212. doi: 10.1007/BF02211841. [DOI] [PubMed] [Google Scholar]
  13. Lotspeich L. J., Ciaranello R. D. The neurobiology and genetics of infantile autism. Int Rev Neurobiol. 1993;35:87–129. doi: 10.1016/s0074-7742(08)60569-3. [DOI] [PubMed] [Google Scholar]
  14. Macpherson J. N., Nelson D. L., Jacobs P. A. Frequent small amplifications in the FMR-1 gene in fra(X) families: limits to the diagnosis of 'premutations'. J Med Genet. 1992 Nov;29(11):802–806. doi: 10.1136/jmg.29.11.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oberlé I., Rousseau F., Heitz D., Kretz C., Devys D., Hanauer A., Boué J., Bertheas M. F., Mandel J. L. Instability of a 550-base pair DNA segment and abnormal methylation in fragile X syndrome. Science. 1991 May 24;252(5009):1097–1102. doi: 10.1126/science.252.5009.1097. [DOI] [PubMed] [Google Scholar]
  16. Payton J. B., Steele M. W., Wenger S. L., Minshew N. J. The fragile X marker and autism in perspective. J Am Acad Child Adolesc Psychiatry. 1989 May;28(3):417–421. doi: 10.1097/00004583-198905000-00019. [DOI] [PubMed] [Google Scholar]
  17. Piven J., Gayle J., Landa R., Wzorek M., Folstein S. The prevalence of fragile X in a sample of autistic individuals diagnosed using a standardized interview. J Am Acad Child Adolesc Psychiatry. 1991 Sep;30(5):825–830. [PubMed] [Google Scholar]
  18. Richards R. I., Holman K., Kozman H., Kremer E., Lynch M., Pritchard M., Yu S., Mulley J., Sutherland G. R. Fragile X syndrome: genetic localisation by linkage mapping of two microsatellite repeats FRAXAC1 and FRAXAC2 which immediately flank the fragile site. J Med Genet. 1991 Dec;28(12):818–823. doi: 10.1136/jmg.28.12.818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ritvo E. R., Jorde L. B., Mason-Brothers A., Freeman B. J., Pingree C., Jones M. B., McMahon W. M., Petersen P. B., Jenson W. R., Mo A. The UCLA-University of Utah epidemiologic survey of autism: recurrence risk estimates and genetic counseling. Am J Psychiatry. 1989 Aug;146(8):1032–1036. doi: 10.1176/ajp.146.8.1032. [DOI] [PubMed] [Google Scholar]
  20. Rousseau F., Heitz D., Biancalana V., Oberlé I., Mandel J. L. On some technical aspects of direct DNA diagnosis of the fragile X syndrome. 1992 Apr 15-May 1Am J Med Genet. 43(1-2):197–207. doi: 10.1002/ajmg.1320430133. [DOI] [PubMed] [Google Scholar]
  21. Saiki R. K., Bugawan T. L., Horn G. T., Mullis K. B., Erlich H. A. Analysis of enzymatically amplified beta-globin and HLA-DQ alpha DNA with allele-specific oligonucleotide probes. Nature. 1986 Nov 13;324(6093):163–166. doi: 10.1038/324163a0. [DOI] [PubMed] [Google Scholar]
  22. Smalley S. L., Asarnow R. F., Spence M. A. Autism and genetics. A decade of research. Arch Gen Psychiatry. 1988 Oct;45(10):953–961. doi: 10.1001/archpsyc.1988.01800340081013. [DOI] [PubMed] [Google Scholar]
  23. Smalley S. L. Genetic influences in autism. Psychiatr Clin North Am. 1991 Mar;14(1):125–139. [PubMed] [Google Scholar]
  24. Snow K., Doud L. K., Hagerman R., Pergolizzi R. G., Erster S. H., Thibodeau S. N. Analysis of a CGG sequence at the FMR-1 locus in fragile X families and in the general population. Am J Hum Genet. 1993 Dec;53(6):1217–1228. [PMC free article] [PubMed] [Google Scholar]
  25. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  26. Spiker D., Lotspeich L., Kraemer H. C., Hallmayer J., McMahon W., Petersen P. B., Nicholas P., Pingree C., Wiese-Slater S., Chiotti C. Genetics of autism: characteristics of affected and unaffected children from 37 multiplex families. Am J Med Genet. 1994 Mar 15;54(1):27–35. doi: 10.1002/ajmg.1320540107. [DOI] [PubMed] [Google Scholar]
  27. Steffen D., Weinberg R. A. The integrated genome of murine leukemia virus. Cell. 1978 Nov;15(3):1003–1010. doi: 10.1016/0092-8674(78)90284-2. [DOI] [PubMed] [Google Scholar]
  28. Steffenburg S., Gillberg C., Hellgren L., Andersson L., Gillberg I. C., Jakobsson G., Bohman M. A twin study of autism in Denmark, Finland, Iceland, Norway and Sweden. J Child Psychol Psychiatry. 1989 May;30(3):405–416. doi: 10.1111/j.1469-7610.1989.tb00254.x. [DOI] [PubMed] [Google Scholar]
  29. Tarleton J., Wong S., Schwartz C. Direct analysis of the FMR-1 gene provides an explanation for an exceptional case of a fragile X negative, mentally retarded male in a fragile X family. J Med Genet. 1992 Dec;29(12):919–920. doi: 10.1136/jmg.29.12.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Verkerk A. J., Pieretti M., Sutcliffe J. S., Fu Y. H., Kuhl D. P., Pizzuti A., Reiner O., Richards S., Victoria M. F., Zhang F. P. Identification of a gene (FMR-1) containing a CGG repeat coincident with a breakpoint cluster region exhibiting length variation in fragile X syndrome. Cell. 1991 May 31;65(5):905–914. doi: 10.1016/0092-8674(91)90397-h. [DOI] [PubMed] [Google Scholar]
  31. Webb T. P., Bundey S., Thake A., Todd J. The frequency of the fragile X chromosome among schoolchildren in Coventry. J Med Genet. 1986 Oct;23(5):396–399. doi: 10.1136/jmg.23.5.396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wehnert M., Reiner O., Caskey C. T. Four STR polymorphisms map to a 500 kb region between DXS15 and DXS134. Hum Mol Genet. 1993 Sep;2(9):1503–1503. doi: 10.1093/hmg/2.9.1503. [DOI] [PubMed] [Google Scholar]
  33. Weissenbach J., Gyapay G., Dib C., Vignal A., Morissette J., Millasseau P., Vaysseix G., Lathrop M. A second-generation linkage map of the human genome. Nature. 1992 Oct 29;359(6398):794–801. doi: 10.1038/359794a0. [DOI] [PubMed] [Google Scholar]

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