Abstract
The lampbrush chromosomes found in the giant nucleus or germinal vesicle (GV) of amphibian oocytes provide unique opportunities for discrete closed and open chromatin structural domains to be directly observable by simple light microscopy. Moreover, the method described here for preparing spreads of lampbrush chromatin for immunostaining enables a straightforward approach to establishing the distributions of modified nucleotides within and between structurally and functionally distinctive chromatin domains.
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References
Callan HG (1986) Lampbrush chromosomes. In: Molecular biology, biochemistry and biophysics, vol 36. Springer-Verlag, Berlin
Gall JG (2014) Transcription in the Xenopus oocyte nucleus. In: Kloc M, Kubiak JZ (eds) Xenopus development. Wiley, Oxford, pp 3–15
Morgan GT (2002) Lampbrush chromosomes and associated bodies: new insights into principles of nuclear structure and function. Chromosom Res 10(3):177–200
Mais C, Scheer U (2001) Molecular architecture of the amplified nucleoli of Xenopus oocytes. J Cell Sci 114:709–718
Sommerville J, Brumwell CL, Politz JC, Pederson T (2005) Signal recognition particle assembly in relation to the function of amplified nucleoli of Xenopus oocytes. J Cell Sci 118:1299–1307
Murphy C, Wang Z, Roeder RG, Gall JG (2002) RNA polymerase III in Cajal bodies and lampbrush chromosomes of the Xenopus oocyte nucleus. Mol Biol Cell 13:3466–3476. https://doi.org/10.1091/mbc.E02-05-0281
Angelier N, Bonnanfant-Jaïs ML, Moreau N, Gounon P, Lavaud A (1986) DNA methylation and RNA transcriptional activity in amphibian lampbrush chromosomes. Chromosoma 94:169–182. https://doi.org/10.1007/bf00288491
Morgan GT, Jones P, Bellini M (2012) Association of modified cytosines and the methylated DNA-binding protein MeCP2 with distinctive structural domains of lampbrush chromatin. Chromosom Res 20:925–942. https://doi.org/10.1007/s10577-012-9324-x
Gall JG, Nizami ZF (2016) Isolation of giant lampbrush chromosomes from living oocytes of frogs and salamanders. J Vis Exp 118(118):e54103. https://doi.org/10.3791/54103
Gall JG, Murphy C, Callan HG, Wu ZA (1991) Lampbrush chromosomes. Methods Cell Biol 36:149–166
Dumont JN (1972) Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals. J Morphol 136:153–179
Beetschen J-C, Gautier J (1989) Oogenesis. In: Armstrong JB, Malacinski GM (eds) Developmental biology of the axolotl. Oxford University Press, New York, Oxford, pp 25–35
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Morgan, G.T. (2021). Immunochemical Detection of Modified Cytosine Species in Lampbrush Chromatin. In: Ruzov, A., Gering, M. (eds) DNA Modifications. Methods in Molecular Biology, vol 2198. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0876-0_13
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DOI: https://doi.org/10.1007/978-1-0716-0876-0_13
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