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First published online 12 February 2008
doi: 10.1242/jcs.025205

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Nuclear organisation in totipotent human nuclei and its relationship to chromosomal abnormality

¹ Department of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
² The London Bridge Fertility, Gynaecology and Genetics Centre, 1 St Thomas Street, London Bridge, London, SE1 9RY, UK
³ Department of Obstetrics and Gynaecology, University College London, 86-96 Chenies Mews, London, WC1E 6HX, UK

View larger version (85K): [in this window] [in a new window]   Fig. 1. Representative image of a human blastomere nucleus probed for loci 13q14 (red), 16q12 (light blue), 18c (dark blue), 21q22 (red) and 22q22.1 (yellow). A transparent five-ringed template (with each shell 1-5 of equal area) is overlaid to determine the relative position of the loci. Scale bar: 5 µm.

View larger version (482K): [in this window] [in a new window]   Fig. 2. The distribution of the eight loci (13q14, 15c, 16q12, 18c, 21q22, 22q11.2, Xc and Yc) in the nuclei of various cell types. The y-axis represents the percentage of signals represented in each shell (1-5) after normalisation to take into account the nuclear counterstain (excluding sperm). Shell 1 (x-axis) represents the outermost shell and shell 5 the innermost (see Fig. 1). Thus, a graph with the highest peak in shell 1 compared with the other shells represents a signal found most often at the nuclear periphery; a graph with the highest peak in shell 5 represents a signal found most often at the nuclear centre. n, the number of signals scored. P values represent the results of chi-squared tests to establish whether a significant non-random pattern could be identified. P<0.05, significantly non-random (P<0.01, highly significant); NOT SIG, not significant (i.e. P>0.05). (A) Normal lymphocyte nuclei. (B) Nuclei of trisomy 13 lymphoblastoid cells. (C) Nuclei of cultured cells from amniotic fluid (chromosomally normal). (D) Sperm-head nuclei (chromosomally normal, fertile male). (E) Blastomere nuclei with no detectable abnormality (NDA group). (F) Aneuploid blastomere nuclei. (G) Chaotic blastomere nuclei. (H) Blastomeres trisomic for the chromosome measured (e.g. position of chromosome-13 locus in trisomy-13 cells).

View larger version (12K): [in this window] [in a new window]   Fig. 3. (A) The distribution of locus 22q11.2 in our data set (see also Fig. 2b). A central location is apparent. (B) The same data set analysed according to McKenzie et al. (McKenzie et al., 2004) – the graph suggests a more medial location. x-axis depicts the shell number.