ABSTRACT
BACKGROUND AND PURPOSE: The heart of the canine model of chronic atrioventricular block is known to have a ventricular electrical remodelling, which mimics the pathophysiology of long QT syndrome. Using this model, we explored a new pharmacological therapeutic strategy for the prevention of cardiac sudden death. EXPERIMENTAL APPROACH: The L-type Ca(2+) channel blocker amlodipine (2.5 mg.day(-1)), L/N-type Ca(2+) channel blocker cilnidipine (5 mg.day(-1)), or the angiotensin II receptor blocker candesartan (12 mg.day(-1)) was administered orally to the dogs with chronic atrioventricular block for 4 weeks. Electropharmacological assessments with the monophasic action potential (MAP) recordings and blood sample analyses were performed before and 4 weeks after the start of drug administration. KEY RESULTS: Amlodipine and cilnidipine decreased the blood pressure, while candesartan hardly affected it. The QT interval, MAP duration and beat-to-beat variability of the ventricular repolarization period were shortened only in the cilnidipine group, but such effects were not observed in the amlodipine or candesartan group. Plasma concentrations of adrenaline, angiotensin II and aldosterone decreased in the cilnidipine group. In contrast, plasma concentrations of angiotensin II and aldosterone were elevated in the amlodipine group, whereas in the candesartan group an increase in plasma levels of angiotensin II and a decrease in noradrenaline and adrenaline concentrations were observed. CONCLUSIONS AND IMPLICATIONS: Long-term blockade of L/N-type Ca(2+) channels ameliorated the ventricular electrical remodelling in the hypertrophied heart which causes the prolongation of the QT interval. This could provide a novel therapeutic strategy for the treatment of cardiovascular diseases.
Subject(s)
Calcium Channel Blockers/pharmacology , Calcium Channels, L-Type/physiology , Calcium Channels, N-Type/physiology , Cardiomegaly/physiopathology , Dihydropyridines/pharmacology , Action Potentials/drug effects , Amlodipine/blood , Amlodipine/pharmacology , Angiotensin II/blood , Angiotensin II Type 1 Receptor Blockers/blood , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Atrioventricular Block/drug therapy , Atrioventricular Block/physiopathology , Benzimidazoles/blood , Benzimidazoles/pharmacology , Biphenyl Compounds , Calcium Channel Blockers/blood , Calcium Channel Blockers/therapeutic use , Chronic Disease , Dihydropyridines/blood , Dihydropyridines/therapeutic use , Dogs , Electrocardiography , Epinephrine/blood , Female , Male , Neurotransmitter Agents/blood , Norepinephrine/blood , Tetrazoles/blood , Tetrazoles/pharmacology , Time FactorsABSTRACT
The imprinted region on mouse distal chromosome 12 covers about 1 Mb and contains at least three paternally expressed genes (Pegs: Peg9/Dlk1, Peg11/Rtl1, and Dio3) and four maternally expressed genes (Megs: Meg3/Gtl2, antiPeg11/antiRlt1, Meg8/Rian, and Meg9/Mirg). Gtl2(lacZ) (Gene trap locus 2) mice have a transgene (TG) insertion 2.3 kb upstream from the Meg3/Gtl2 promoter and show about 40% growth retardation when the TG-inserted allele is paternally derived. Quantitative RT-PCR experiments showed that the expression levels of Pegs in this region were reduced below 50%. These results are consistent with the observed phenotype in Gtl2lacZ mice, because at least two Pegs(Peg9/Dlk1 and Dio3) have growth-promoting effects. The aberrant induction of Megs from silent paternal alleles was also observed in association with changes in the DNA methylation level of a differentially methylated region (DMR) located around Meg3/Gtl2 exon 1. Interestingly, a 60 approximately 80% reduction in all Megs was observed when the TG was maternally derived, although the pups showed no apparent growth or morphological abnormalities. Therefore, the paternal or maternal inheritance of the TG results in the down-regulation in cis of either Pegs or Megs, respectively, suggesting that the TG insertion influences the mechanism regulating the entire imprinted region.
Subject(s)
Genomic Imprinting , Proteins/genetics , Animals , Base Sequence , Chromosome Aberrations , Chromosome Mapping , DNA Primers , Female , Gene Expression Regulation , Growth Disorders/genetics , Male , Mice , Mice, Transgenic , Mutagenesis, Insertional , RNA, Long Noncoding , Reverse Transcriptase Polymerase Chain Reaction , beta-Galactosidase/geneticsABSTRACT
A novel paternally expressed imprinted gene, PEG10 (Paternally Expressed 10), was identified on human chromosome 7q21. PEG10 is located near the SGCE (Sarcoglycan epsilon) gene, whose mouse homologue was recently shown to be imprinted. Therefore, it is highly possible that a new imprinted gene cluster exists on human chromosome 7q21. Analysis of two predicted open reading frames (ORF1 and ORF2) revealed that ORF1 and ORF2 have homology to the gag and pol proteins of some vertebrate retrotransposons, respectively. These data suggest that PEG10 is derived from a retrotransposon that was previously integrated into the mammalian genome. PEG10 is likely to be essential for understanding how exogenous genes become imprinted.
Subject(s)
Chromosomes, Human, Pair 7 , Genomic Imprinting , Proteins/genetics , Retroelements , Amino Acid Sequence , Animals , Apoptosis Regulatory Proteins , Choriocarcinoma/genetics , DNA-Binding Proteins , Female , Genes, gag/genetics , Genes, pol/genetics , Humans , Male , Mice , Molecular Sequence Data , Nuclear Proteins/genetics , Open Reading Frames , Physical Chromosome Mapping/methods , Polymorphism, Genetic , RNA-Binding Proteins , Radiation Hybrid Mapping/methods , Sequence Homology, Amino Acid , Syndrome , Transcription Factors/geneticsABSTRACT
BACKGROUND: The paternal duplication of mouse distal chromosome 12 leads to late embryonal/neonatal lethality and growth promotion, whereas maternal duplication leads to late embryonal lethality and growth retardation. Human paternal or maternal uniparental disomies of chromosome 14q that are syntenic to mouse distal chromosome 12 have also been reported to show some imprinting effects on growth, mental activity and musculoskeletal morphology. For the isolation of imprinted genes in this region, a systematic screen of maternally expressed genes (Megs) was carried out by our subtraction-hybridization method using androgenetic and normally fertilized embryos. RESULTS: We have isolated seven candidate clones of the mouse Meg gene. Among them, we identified a novel maternally expressed imprinted gene, Meg3, on mouse distal chromosome 12 and showed that it was identical to the Gtl2 gene. We also found that the human homologue MEG3 on chromosome 14q was also monoallelically expressed. CONCLUSIONS: This is the first identification of the imprinting gene, both on mouse distal chromosome 12 and on human chromosome 14q, respectively. Because there are no obvious open reading frames in either the mouse Meg3/Gtl2 or human MEG3, the function of these genes remains unclear. However, this result will provide a good basis for the further investigation of several important imprinted genes in this chromosomal region.
Subject(s)
Chromosomes, Human, Pair 14 , Genome, Human , Genomic Imprinting , Animals , Chromosome Mapping , Genome , Humans , MiceABSTRACT
BACKGROUND: Genomic imprinting significantly influences development, growth and behaviour in mammals. Systematic screening of imprinted genes has been extensively carried out to identify the genes responsible for imprinted phenotypes and to elucidate the biological significance of this phenomenon. In this study, we applied DNA chip technology for isolating paternally expressed imprinted genes (Pegs). We compared the resulting expression profiles of parthenogenetic and fertilized control embryos to identify novel imprinted genes. RESULTS: A novel paternally expressed mouse imprinted gene, Peg9/Dlk1, was identified. Consistent with this finding, the paternal expression of its human homologue, PEG9/DLK1, was also confirmed. These two genes form imprinted gene clusters with the reciprocally imprinted mouse Meg3/Gtl2 and human MEG3 genes that we first identified on distal chromosome 12 and chromosome 14q32, respectively. CONCLUSIONS: As DNA chip technology allows us to quickly screen a large number of genes, using this technology to search for imprinted genes could accelerate the identification of genes responsible for human and mouse genetic diseases. Dlk1 and DLK1, which encode transmembrane proteins, have six EGF-like repeats and show homology to the Delta gene in Drosophila melanogaster. Because of its homology to mammalian Delta homologues, PEG9/DLK1 may contribute to the scoliosis phenotype observed in maternal uniparental disomy 14 (mUPD14) patients.
Subject(s)
Gene Expression Regulation, Developmental , Gene Order , Genomic Imprinting , Membrane Proteins/genetics , Proteins/genetics , Animals , Crosses, Genetic , Embryo, Mammalian , Female , Gene Expression Profiling , Humans , Intracellular Signaling Peptides and Proteins , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Parthenogenesis , RNA, Long NoncodingABSTRACT
We developed a visual data analysis system that can easily manage a large volume of medical imaging data. This system can analyze sets of imaging data using general image processing methods, so that various kinds of medical imaging data such as ECG charts, X ray image films, and MRI images, can be processed. The system has a graphical user interface (GUI). A physician who is novice at the system can manipulate the imaging data intuitively by pull down menus, pop up menus and buttons within the window system. The system can run on a standard UNIX workstation which is faster and more powerful than most personal computers. The system needs an X window system/Motif and C compiler. These are standard system programs already available on most UNIX workstations. The source code of the system can be retrieved from our anonymous ftp site via Internet.
