30 research outputs found
Clinical data of probands and relatives carrying a large <i>PKP2</i> heterozygous deletion.
<p>Clinical data of probands and relatives carrying a large <i>PKP2</i> heterozygous deletion.</p
Detailed list and characteristics of putative mutations.
<p>Detailed list and characteristics of putative mutations.</p
Pedigree and qPCR results from family A and U.
<p>qPCR showed a ~50% reduction of exon 4 <i>PKP2</i> DNA in affected individuals A.1, A.2 and A3 compared to the healthy father A.4 (used as a control). <i>PKP2</i> DNA quantification was normal in exon 3 and 5 for all individuals. In family U, qPCR showed a 50% reduction of <i>PKP2</i> DNA in exon 4 and 13 in affected individuals U.1 and U.2 and their mother U.3 whereas it was comparable to controls in father U.4. Individuals U.1 and U.2 also carried the p.Gly712Arg (G712R) rare <i>PKP2</i> variant that falsely appeared in the Sanger chromatogram as homozygous (HmZ) instead of hemizygous because of the absence of the other <i>PKP2</i> allele. This variant was inherited from the father, in absence of familial history of consanguinity, in whom it appeared heterozygous(Htz).</p
Clinical characteristics of patients with <i>PCDH19</i> mutations.
<p>2*: patient N 06 1258 is the sister of patient N 06 1257 (index case); PMD = psychomotor development, Nl = normal, F = febrile, unit = unilateral, GTC = generalized tonic-clonic, W-S = words-sentences, abs = absent, AED = anti epileptic drugs.</p
Detection of 9 different point mutations of <i>PCDH19</i> in 11 female patients by direct sequencing.
<p>A) Sequence electropherograms of the mutations and the missense variant (c.3319C>G/p.Arg1107Gly) identified in association with the c.859G>T/p.Glu287X nonsense mutation. The mutation nomenclature is based on the <i>PCDH19</i> transcript reference EF676096. Nucleotides are numbered according to the cDNA with +1 corresponding to the A of the ATG translation initiation codon in the reference sequence, according to the journal guidelines (<a href="http://www.hgvs.org/mutnomen" target="_blank">www.hgvs.org/mutnomen</a>). B) Alignment of the regions surrounding the mutations (indicated by an arrow) in orthologous and paralogous proteins, showing the high conservation of each affected amino-acid in vertebrates and in the delta protocadherin paralogous genes.</p
FISH analysis of the <i>PCDH19</i> deletion in the male patient showing somatic mosaicism in fibroblasts.
<p>(A) Absence of the specific Xq22.1 probe site on metaphase chromosomes in peripheral blood lymphocytes (PBL); (B) In fibroblasts, presence of one hybridization spot in 53% of the cells and absence of signal in the remaining 47%; C) and D) FISH analysis on PBL (C) and fibroblasts (D) of a female control. <i>PCDH19</i>-specific signals (red) are indicated by arrowheads. Magnification Ă—1000.</p
Schematic illustration of the cellular interference mechanism associated with <i>PCDH19</i> mutations.
<p>A) In normal individuals, characterized by a homogeneous population of <i>PCDH19</i>-positive cells, neurons are able to form normal neuronal networks; B) In mutated male patients, hemizygosity leads to a homogeneous population of <i>PCDH19</i>-negative cells; in this condition, neurons preserve the ability to form normal neuronal networks; C) In heterozygous mutated females, random X inactivation leads to the co-existence of two <i>PCDH19</i>-positive and <i>PCDH19</i>-negative cell populations. These two cell populations cause divergent cell sorting and migration (due to attractive or repulsive interactions) and lead to abnormal neuronal networks. Somatic mosaicism in mutated males gives rise to the same pathological situation. The precise mechanisms by which the neuronal networks are altered are still unknown.</p
Pedigrees of the families and segregation analysis of the <i>PCDH19</i> deletion and point mutations.
<p>del/+, m/+ or v/+ denote individuals heterozygous for the deletion, mutation or variant, respectively; +/+ denotes individuals carrying homozygous wild-type alleles. Squares represent males, circles females; filled black symbols: patients diagnosed as having Dravet syndrome; right black half: Cognitive delay or impairment; left grey half: adolescence-onset idiopathic epilepsy. Dots in the middle of the squares indicate unaffected mutation carriers. The arrows indicate the index cases.</p
Garland nephrocyte phenotype of <i>hts</i><sup><i>null</i></sup> and adducin-αγ rescue mutants.
<p>(A) Kirre and Pyd localization in <i>hts</i><sup><i>null</i></sup> and rescue mutant garland nephrocytes. Dissected nephrocytes of the indicated genotypes were stained for Kirre (red) and Pyd, corresponding to Neph1 and ZO-1 in vertebrates, (blue). Arrowheads show areas of cell fusion. Scale bar: 10μm. (B) Quantification of nephrocytes showing a continuous Kirre staining using >9 samples/genotype from 3 independent experiments. Statistical analysis was performed with Kruskal-Wallis with Dunn’s post-test. ns, non significant, *p<0.05, ***p<0.001 (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007386#pgen.1007386.s009" target="_blank">S1 Table</a> for details on transgenic flies). (C) Pericardial nephrocytes in adducin-αγ WT and E559Q rescue and control adult flies at 15 days post-eclosion were stained for the differentiation markers Kirre (red) and Pyd (blue). Note that <i>hts</i><sup><i>null</i></sup> is lethal at this stage. Scale bar: 30μm. (D) Quantification of the number of pericardial nephrocytes from n>8 samples/genotype in 3 independent experiments. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post-test. ns, non significant (See <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007386#pgen.1007386.s009" target="_blank">S1 Table</a> for details on transgenic flies).</p