11.Hypoxic ventilatory depressionと思われる一症例について(第551回千葉医学会例会・第9回麻酔科例会・第18回千葉麻酔懇話会)

FISH analysis on metaphase nuclei (top panel) of cultured cells derived from peripheral blood leukocytes of the proband of family 2 by using BAC probes for 11p15.5-15.4 (RP11-11A9, 3,236,552-3,356,012, green) and 11q22.3 (RP11-179B7, 104,298,339-104,459,797, red). The green signal on both homologues is visible only at chr11p, demonstrating the presence of an in cis duplication and excluding an unbalanced translocation. FISH analysis on interphase nuclei (bottom panel) using the BACs RP11-699D10 (2.9–3.0 Mb, red) and RP11-11A9 (green), hybridizing within the duplication. Note that single and duplicated signals can be seen on the two homologues, respectively. The red-green-green-red order of the duplicated signals indicates that the duplication is inverted. (PDF 52 kb

Additional file 1: of Is ZFP57 binding to H19/IGF2:IG-DMR affected in Silver-Russell syndrome?

Table S1. List of CTCF, OCT4/SOX2 and ZFP57 binding regions and potential target sites in the H19/IGF2:IG-DMR. Genomic positions of the binding regions (if demonstrated) and potential target sites and motifs of all these factors have been listed. (PDF 86 kb

Additional file 6: Figure S6. of Two maternal duplications involving the CDKN1C gene are associated with contrasting growth phenotypes

DNA methylation (top) and copy number (CN, bottom) analyses at 11p15 region in family 2, determined by MS-MLPA. The histograms represent the normalized DNA methylation of ICR1 and ICR2 and CN of the genomic region spanning from the NSD1 to KCNQ1 gene. The CN range that is considered normal is shadowed. Note that methylation of ICR2 is abnormally low while CN values of ICR2 and KCNQ1 exon 13-17 are abnormally high in the proband and his mother. (PDF 130 kb

Additional file 1: Figure S1. of Two maternal duplications involving the CDKN1C gene are associated with contrasting growth phenotypes

Comparative genomic hybridization analysis performed on DNA of the proband from family 1. The extension, the genomic localization (GRC h37/hg19) and the genes included in the duplication are shown. (PDF 54 kb

Additional file 7: Figure S7. of Two maternal duplications involving the CDKN1C gene are associated with contrasting growth phenotypes

Note that the haplotype associated with the duplication segregates from I-1 to II-2 and III-1 and that only the D11S4088 marker shows allelic imbalances in II-2 and III-1. (PDF 141 kb

Additional file 2: Figure S2. of Two maternal duplications involving the CDKN1C gene are associated with contrasting growth phenotypes

Single nucleotide polymorphism analysis of genomic DNA from relatives of family 1. I-1 = maternal grandfather, II-2 = mother, III-1 = first brother of the proband, and III-2 = second brother of the proband. Note that the duplication is present in II-2 and III-2 but not in I-1 and III1. (PDF 127 kb

Additional file 4: Figure S4. of Two maternal duplications involving the CDKN1C gene are associated with contrasting growth phenotypes

FISH analysis on metaphase nuclei (top panel) of cultured cells derived from the umbilical cord of the proband of family 1 by using BAC probes for 11p15.5-15.4 (RP11-81K4, 2798699-2970438, green) and 11q22.3 (RP11-876C12, 103,804,669-103,982,517, red). The green signal on both homologues is visible only at chr11p, demonstrating the presence of an in cis duplication and excluding an unbalanced translocation. FISH analysis on interphase nuclei (bottom panel) using the BACs RP11-11A9 (3,236,552-3,356,012, green) and RP11-81K4 (red), hybridizing within the duplication. Note that single and duplicated signals can be seen on the two homologues, respectively. The green-red-red-green order of the duplicated signals indicates that the duplication is inverted. (PDF 51 kb

Tissue-specific and mosaic imprinting defects underlie opposite congenital growth disorders in mice

<div><p>Differential DNA methylation defects of <i>H19/IGF2</i> are associated with congenital growth disorders characterized by opposite clinical pictures. Due to structural differences between human and mouse, the mechanisms by which mutations of the <i>H19/IGF2</i> Imprinting Control region (IC1) result in these diseases are undefined. To address this issue, we previously generated a mouse line carrying a humanized IC1 (hIC1) and now replaced the wildtype with a mutant IC1 identified in the overgrowth-associated Beckwith-Wiedemann syndrome. The new humanized mouse line shows pre/post-natal overgrowth on maternal transmission and pre/post-natal undergrowth on paternal transmission of the mutation. The mutant hIC1 acquires abnormal methylation during development causing opposite <i>H19/Igf2</i> imprinting defects on maternal and paternal chromosomes. Differential and possibly mosaic <i>Igf2</i> expression and imprinting is associated with asymmetric growth of bilateral organs. Furthermore, tissue-specific imprinting defects result in deficient liver- and placenta-derived <i>Igf2</i> on paternal transmission and excessive <i>Igf2</i> in peripheral tissues on maternal transmission, providing a possible molecular explanation for imprinting-associated and phenotypically contrasting growth disorders.</p></div

Somatic undergrowth on paternal transmission of the <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2</i>} allele.

<p>(A-C) Box plot of birth weights (A), growth charts (B), and box plots of organ weights at 14 weeks of age (C) of <i>H19</i>^{<i>+/hIC1</i>Δ<i>2</i>.<i>2</i>} and <i>H19</i>^<i>+/+</i> littermates. Box plots in (A) and (C) and growth chart in (B) are as in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007243#pgen.1007243.g002" target="_blank">Fig 2</a>. (D-E) Box plot of embryo body and placenta weights of <i>H19</i>^{<i>+/hIC1</i>Δ<i>2</i>.<i>2</i>} (D) and <i>H19</i>^{<i>+/hIC1</i>} (E) mice at E15.5 compared with <i>H19</i>^<i>+/+</i> littermates. The animals used for this study derived from three (A-C) or two litters (D-E).</p

Gain of IC1 methylation in <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2/+</i>} mice.

<p>(A) Graph representing the humanized and endogenous <i>H19/Igf2</i> locus with the regions (indicated by black bars) whose methylation was analysed by pyrosequencing. The region deleted in <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2</i>} is represented by a shaded light blue box. The coordinates of the sequenced CpGs are: CTS1 in hg39—chr11:2,024,193–2,024,261; CTS6 in hg39—chr11:2,021,096–2,021,153; mIC1 in mm9—chr7:149,767,625–149,767,705; mIC2 in mm9—chr7:150,482,365–150,482,486. Other details are described in the legend to <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007243#pgen.1007243.g001" target="_blank">Fig 1</a>. (B) Percent methylation measured by pyrosequencing at two CTCF target sites (CTS1 and CTS6) in three different organs collected from <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2/+</i>} and <i>H19</i>^{<i>hIC1/+</i>} mice at birth (left panel). Endogenous mIC1 analysed as control of methylation: the paternal allele (pat) was assayed in <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2/+</i>}, the maternal allele (mat) in <i>H19</i>^{<i>+/hIC1</i>Δ<i>2</i>.<i>2</i>}, and both alleles (pat + mat) in <i>H19</i>^<i>+/+</i> mice (right panel). Each histogram represents the methylation mean value of 5 (CTS1 and mIC1) or 6 (CTS6) CpGs, tested in <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2/+</i>} (n = 10), <i>H19</i>^{<i>hIC1/+</i>} (n = 9) and <i>H19</i>^<i>+/+</i> (n = 7) mice derived from three litters. Bars represent the mean ± SEM. Note that methylation (ranging from 55% to 75%) was observed at the <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2</i>} and not at the <i>H19</i>^<i>hIC1</i> allele. (C) IC1 methylation analysed by bisulphite treatment followed by cloning and sequencing in tongue of <i>H19</i>^{<i>hIC1/+</i>} and <i>H19</i>^{<i>hIC1</i>Δ<i>2</i>.<i>2/+</i>} mice. Each line corresponds to a single template DNA molecule cloned; each circle corresponds to a CpG dinucleotide. Filled circles designate methylated cytosines; open circles, unmethylated cytosines. CpGs measured by pyrosequencing are framed.</p