Abstract
For the cytogenetic study of X chromosome inactivation as an X chromosome dosage compensation mechanism, we isolated a number of XXXX, XXX, and XXY near-tetraploid mouse hybrid cell clones by fusing XX or XO embryonal carcinoma cells with lymphocytes carrying a structurally altered X chromosome(s). The inactive X chromosome from the female lymphocyte was reactivated in these hybrid clones which retained embryonal carcinoma morphology so far as they were cultured on the collagen-coated plastic surface in the medium supplemented with leukemia inhibitory factor (LIF) and betamercaptoethanol (BME). Some of these clones developed balloon-like cystic embryoid bodies when they were allowed to form cell aggregates in medium without LIF and BME in bacteriological petri dishes to which they do not adhere. X chromosome inactivation occurring during this process detected by the incorporation of 5-bromodeoxyuridine did not conform to the expected pattern leaving two X chromosomes active in every tetraploid cells. This may suggest either that the X-inactivation mechanism evolved primarily, for the diploid cell is unable to deal with tetraploid conditions efficiently, or that the present system ofin vitro differentiation represents an anomalous situation never encounteredin vivo.
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Adler, I.-D., R. Johannisson & H. Winking, 1989. The influence of Robertsonian translocation Rb(X.2)2Ad on anaphase I non-disjunction in male laboratory mice. Genet. Res. 52: 77–86.
Andrews, P. W. & P. N. Goodfellow, 1980. Antigen expression by somatic cell hybrids of a murine embryonal carcinoma cell with thymocytes and L cells. Somatic Cell Genet. 6: 271–284.
Cattanach, B. M., 1975. Control of chromosome inactivation. Annu. Rev. Genet. 9: 1–18.
Drews, U., St. R. Blecher, D. A. Owen & S. Ohno, 1974. Genetically directed preferential X-inactivation seen in mice. Cell 1: 3–8.
Gartler, S. M. & A. D. Riggs, 1983. Mammalian X-chromosome inactivation. Annu. Rev. Genet. 17: 155–190.
Grant, S. G. & V. M. Chapman, 1988. Mechanisms of X-chromosome regulation. Annu. Rev. Genet. 22: 199–233.
Hales, A., 1977. A procedure for the fusion of cells in suspension by means of polyethylene glycol. Somatic Cell Genet. 3: 227–230.
Hamerton, J. L., 1971. Human Cytogenetics. Academic Press, New York & London.
Hogan, B., F. Costantini & E. Lacy, 1986. Manipulating the mouse embryo. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Lyon, M. F., 1961. Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature 190: 372–373.
Lyon, M. F. & S. Rastan, 1984. Parental source of chromosome imprinting and its relevance for X chromosome inactivation. Differentiation 26: 63–67.
Lyon, M. F., J. Zenthon, E. P. Evans, M. D. Burtenshaw, K. A. Wareham & E. D. Williams, 1986. Lack of inactivation of a mouse X-linked gene physically separated from the inactivation centre. J. Embryol. exp. Morph. 97: 75–85.
Lyon, M. F., 1988. The William Allan Memorial Award Address: X-chromosome inactivation and the location and expression of X-linked genes. Am. J. Hum. Genet. 42: 8–16.
Martin, G. R., C. J. Epstein, B. Travis, G. Tucker, S. Yatsiv, D. W. Martin Jr, S. Clift & S. Cohen, 1978. X-chromosome inactivation during differentiation of female teratocarcinoma stem cellsin vitro. Nature 271: 329–333.
Martin, G. R., L. M. Wiley & I. Damjanov, 1977. The development of cystic embryoid bodies in vitro from clonal teratocarcinoma stem cells. Dev. Biol. 61: 230–244.
Martin, G. R., 1980. Teratocarcinomas and mammalian embryogenesis. Science 209: 768–772.
McBurney, M. W. & B. J. Strutt, 1980. Genetic activity of the X chromosomes in pluripotential teratocarcinoma cells and their differentiated progeny. Cell 21: 357–364.
Miller, R. A. & F. H. Ruddle, 1976. Pluripotent teratocarcinoma-thymus somatic cell hybrids. Cell 9: 45–55.
Miller, R. A. & F. H. Ruddle, 1977. Properties of teratocarcinoma-thymus somatic cell hybrids. Somatic Cell Genet. 3: 247–261.
Okuyama, K., N. Takagi & M. Sasaki, 1986. Sequential X-chromosome reactivation and inactivation in cell hybrids between murine embryonal carcinoma cells and female rat thymocytes. Exp. Cell Res. 164: 323–334.
Paterno, G. D. & M. W. McBurney, 1985. X chromosome inactivation during induced differentiation of a female mouse embryonal carcinoma cell line. J. Cell Sci. 75: 149–163.
Rastan, S., 1983. Nonrandom X chromosome inactivation in mouse X-autosome translocation embryos — location of the inactivation centre. J. Embryol. exp. Morph. 78: 1–22.
Rousset, J.-P., J. Jami, P. Dubois, D. Aviles & E. Rits, 1980. Developmental potentialities and surface antigens of mouse teratocarcinoma x lymphoid cell hybrids. Somatic Cell Genet. 6: 419–433.
Russell, L. B., 1983. X-autosome translocations in the mouse: Their characterization and use as tools to investigate gene inactivation and gene action. In: A. A. Sandberg (ed.). Cytogenetics of the mammalian X-chromosome. Part. A. Basic mechanisms of X-chromosome behavior. A. R. Liss, New York: 205–250.
Searle, A. G., C. V. Beechey, E. P. Evans & M. Kirk, 1983. Two new A-autosome translocations in the mouse. Cytogenet. Cell Genet. 35: 279–291.
Shao, C. & N. Takagi, 1990. An extra maternally derived X chromosome is deleterious to early mouse development. Development 110: 969–975.
Schindler, A. M. & K. Mikamo, 1970. Triploidy in man: Report of a case and a discussion on etiology. Cytogenetics 9: 116–130.
Snow, M. H. L., 1975. Embryonic development of tetraploid mice during the second half of gestation. J. Embryol. exp. Morph. 34: 707–721.
Takagi, N. & M. Sasaki, 1975. Preferential inactivation of the paternally derived X chromosome in the extraembryonic membranes of the mouse. Nature 256: 640–642.
Takagi, N., 1983. De novo X-chromosome inactivation in somatic hybrid cells between the XO mouse embryonal carcinoma cells and XY rat lymphocytes. Exp. Cell Res. 145: 397–404.
Takagi, N., M. A. Yoshida, O. Sugawara & M. Sasaki, 1983. Reversal of X-inactivation in female mouse somatic cells hybridized with murine teratocarcinoma stem cells in vitro. Cell 34: 1053–1062.
Takagi, N. & G. R. Martin, 1984. Studies on the temporal relationship between the cytogenetic and biochemical manifestations of X-chromosome inactivation during the differentiation of LT-1 teratocarcinoma stem cells. Dev. Biol. 103: 425–433.
Takagi, N., 1988. Requirement of mitoses for the reversal of X-inactivation in cell hybrids between murine embryonal carcinoma cells and normal female thymocytes. Expo. Cell Res. 175: 363–373.
Takagi, N. & K. Abe, 1990. Detrimental effects of two active X chromosomes on early mouse development. Development 109: 189–201.
Takagi, N., 1991. Abnormal X-chromosome dosage compensation as a possible cause of early developmental failure in mice. Develop, Growth Differ. 33: 429–435.
Webb, S., T. J. de Vries & M. H. Kaufman, 1992. The differential staining pattern of the X chromosome in the embryonic and extraembryonic tissues of postimplantation homozygous tetraploid mouse embryos. Genet. Res. 59: 205–214.
West, J. D., W. I. Frels, V. M. Chapman & V. E. Papaioannou, 1977. Preferential expression of the maternally derived X-chromosome in the mouse yolk sac. Cell 12: 873–882.
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Takagi, N. Variable X chromosome inactivation patterns in near-tetraploid murine EC × somatic cell hybrid cells differentiatedin vitro . Genetica 88, 107–117 (1993). https://doi.org/10.1007/BF02424467
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DOI: https://doi.org/10.1007/BF02424467