RESUMO
BACKGROUND: Evaluation of gene expression in failing human heart has been limited by the availability of cardiac tissue. METHODS AND RESULTS: We used the polymerase chain reaction (PCR) to assess gene expression in small quantities of failing and nonfailing human heart. PCR is a powerful new molecular biological tool that allows a small quantity of DNA to be amplified as much as 1 million-fold. Total RNA was extracted from 3-5 mg samples of human heart and reverse-transcribed to complementary DNA (cDNA). With selected oligonucleotide primers, we used PCR to amplify cDNAs encoding atrial natriuretic peptide, beta-myosin heavy chain, phospholamban, and cytoskeletal beta-actin. To quantify the relative levels of messenger RNA (mRNA) in human heart, a known amount of a control RNA was present in the reverse transcription and PCR reactions. The amount of mRNA in the sample could therefore be assessed in relation to the amount of control product. The control RNA was transcribed from a synthetic DNA template containing primers complementary to those used to amplify the cDNAs of interest. Atrial natriuretic factor mRNA could not be detected in nonfailing human heart but was abundant in ventricular myocardium from failing human heart. In contrast, steady-state levels of phospholamban mRNA decreased, whereas levels of beta-myosin heavy-chain mRNA were unchanged with heart failure. CONCLUSIONS: Alterations in gene expression in the failing human heart appear to be selective. In addition, the present study suggests that PCR provides a rapid and economical way to quantify the expression of multiple genes of interest in endomyocardial biopsy specimens and may therefore be used to advance our understanding of heart muscle disease.
Assuntos
Baixo Débito Cardíaco/genética , Endocárdio/metabolismo , Regulação da Expressão Gênica , Miocárdio/metabolismo , RNA Mensageiro/metabolismo , Biópsia , Baixo Débito Cardíaco/metabolismo , Cardiomiopatia Dilatada/metabolismo , Cardiomiopatia Dilatada/patologia , DNA/metabolismo , Endocárdio/patologia , Humanos , Miocárdio/patologia , Concentração Osmolar , Reação em Cadeia da Polimerase , EspectrofotometriaRESUMO
The gene for the type I interleukin-1 (IL-1) receptor has been mapped in both mouse and human. In the human genome, a combination of segregation analysis of rodent-human hybrid cells and chromosomal in situ hybridization has placed the gene on the long arm of chromosome 2, at band 2q12. This is near the reported map position of the loci for IL-1 alpha and IL-1 beta (2q13----2q21). The murine gene has been mapped by analysis of restriction fragment length polymorphisms in interspecific backcrosses to the centromeric end of chromosome 1, in a region that is syntenic to a portion of human chromosome 2. The murine Il-1r1 gene has thus been separated from the IL-1 genes, which lie on murine chromosome 2.
Assuntos
Cromossomos Humanos Par 2 , Interleucina-1/metabolismo , Receptores Imunológicos/genética , Animais , Mapeamento Cromossômico , Cruzamentos Genéticos , Genes , Ligação Genética , Humanos , Células Híbridas , Camundongos , Hibridização de Ácido Nucleico , Polimorfismo de Fragmento de Restrição , Receptores Imunológicos/metabolismo , Receptores de Interleucina-1 , Recombinação GenéticaRESUMO
Interspecific mouse backcross analysis was used to generate a molecular genetic linkage map of mouse chromosome 10. The map locations of the Act-2, Ahi-1, Bcr, Braf, Cdc-2a, Col6a-1, Col6a-2, Cos-1, Esr, Fyn, Gli, Ifg, Igf-1, Myb, Pah, pgcha, Ros-1 and S100b loci were determined. These loci extend over 80% of the genetic length of the chromosome, providing molecular access to many regions of chromosome 10 for the first time. The locations of the genes mapped in this study extend the known regions of synteny between mouse chromosome 10 and human chromosomes 6, 10, 12 and 21, and reveal a novel homology segment between mouse chromosome 10 and human chromosome 22. Several loci may lie close to, or correspond to, known mutations. Preferential transmission of Mus spretus-derived alleles was observed for loci mapping to the central region of mouse chromosome 10.
Assuntos
Mapeamento Cromossômico , Cruzamentos Genéticos , Ligação Genética , Marcadores Genéticos , Animais , Southern Blotting , Cromossomos Humanos , Sondas de DNA , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Polimorfismo de Fragmento de Restrição , Homologia de Sequência do Ácido NucleicoRESUMO
We have generated a 30-cM molecular genetic linkage map of the proximal half of mouse chromosome 14 by interspecific backcross analysis. Loci that were mapped in this study include Bmp-1, Ctla-1, Hap, hr, Plau, Psp-2, Rib-1, and Tcra. A region of homology between mouse chromosome 14 and human chromosome 10 was identified by the localization of Plau to chromosome 14. This interspecific backcross map will be valuable for establishing linkage relationships of additional loci to mouse chromosome 14.
Assuntos
Muridae/genética , Animais , Mapeamento Cromossômico , Cromossomos Humanos Par 10 , Cruzamentos Genéticos , Ligação Genética , Marcadores Genéticos , Humanos , Camundongos , Camundongos Endogâmicos C57BL/genética , Polimorfismo de Fragmento de Restrição , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
Interspecific backcross mice were used to create a molecular genetic linkage map of chromosome 2. Genomic DNAs from N2 progeny were subjected to Southern blot analysis using molecular probes that identified the Abl, Acra, Ass, C5, Cas-1, Fshb, Gcg, Hox-5.1, Jgf-1, Kras-3, Ltk, Pax-1, Prn-p, and Spna-2 loci; these loci were added to the 11 loci previously mapped to the distal region of chromosome 2 in the same interspecific backcross to generate a composite multilocus linkage map. Several loci mapped near, and may be the same as, known mutations. Comparisons between the mouse and the human genomes indicate that mouse chromosome 2 contains regions homologous to at least six human chromosomes. Mouse models for human diseases are discussed.
Assuntos
Mapeamento Cromossômico , Muridae/genética , Animais , Cruzamentos Genéticos , Modelos Animais de Doenças , Ligação Genética , Humanos , Camundongos , Camundongos Endogâmicos C57BL/genética , Camundongos Mutantes/genética , Oncogenes , Polimorfismo de Fragmento de Restrição , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
Chromosomal locations have been assigned to seven members of the TGF-beta superfamily using an interspecific mouse backcross. Probes for the Tgfb-1, -2, and -3, Bmp-2a and -3, and Vgr-1 genes recognized only single loci, whereas the Bmp-2b probe recognized two independently segregating loci (designated Bmp-2b1 and Bmp-2b2). The results show that the seven members of the TGF-beta superfamily map to eight different chromosomes, indicating that the TGF-beta family has become widely dispersed during evolution. Five of the eight loci (Tgfb-1, Bmp-2a, Bmp-2b1, Bmp-2b2, Vgr-1) mapped near mutant loci associated with connective tissue and skeletal disorders, raising the possibility that at least some of these mutations result from defects in TGF-beta-related genes.
Assuntos
Camundongos Mutantes/genética , Família Multigênica , Muridae/genética , Fatores de Crescimento Transformadores/genética , Sequência de Aminoácidos , Animais , Mapeamento Cromossômico , Cruzamentos Genéticos , Genes , Ligação Genética , Humanos , Camundongos , Camundongos Endogâmicos/genética , Dados de Sequência Molecular , Polimorfismo de Fragmento de Restrição , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
A molecular genetic linkage map of mouse chromosome 13 was constructed using cloned DNA markers and interspecific backcross mice from two independent crosses. The map locations of Ctla-3, Dhfr, Fim-1, 4/12, Hexb, Hilda, Inhba, Lamb-1.13, Ral, Rrm2-ps3, and Tcrg were determined with respect to the beige (bg) and satin (sa) loci. The map locations of these genes confirm and extend regions of homology between mouse chromosome 13 and human chromosomes 5 and 7, and identify a region of homology between mouse chromosome 13 and human chromosome 6. The molecular genetic linkage map of chromosome 13 provides a framework for establishing linkage relationships between cloned DNA markers and known mouse mutations and for identifying homologous genes in mice and humans that may be involved in disease processes.
Assuntos
Mapeamento Cromossômico , Ligação Genética , Animais , Cromossomos/ultraestrutura , Clonagem Molecular , Cruzamentos Genéticos , Genes , Camundongos , Camundongos Endogâmicos C57BL , FenótipoRESUMO
The zona pellucida is a unique, oocyte-specific matrix that coats the surface of all mammalian eggs. Composed of three sulfated glycoproteins in the mouse (ZP1, ZP2, and ZP3), the zona pellucida facilitates early events in fertilization and protects the embryo during preimplantation development. Using DNA isolated from hamster-mouse somatic cell hybrids and from C57BL/6J X Mus spretus interspecific backcross progeny, Zp-2 was located on chromosome 7, 11.3 +/- 3.2 cM distal to Tyr, and Zp-3 was located on chromosome 5, 9.2 +/- 2.9 cM distal to Gus.
