RESUMEN
An improved linkage map for rat Chromosome (Chr) 10 with two F2 populations was constructed. Thirty new microsatellite markers were generated from a Chr 10-specific, small-insert genomic library and mapped to rat Chr 10. Among them were the rat homologs for the mouse gene for light and heavy chains of myeloperoxidase and human neurofibromatosis 1. Eight newly generated markers (D10Mco62, D10Mco63, D10Mco64, D10Mco65, D10Mco67, D10Mco68, D10Mco70, and D10Mco74) were mapped to the region of the rat Chr 10 blood pressure QTL. The availability of such markers may be instrumental in the search for genes responsible for the hypertension.
Asunto(s)
Ligamiento Genético , Repeticiones de Microsatélite , Animales , Mapeo Cromosómico , Datos de Secuencia Molecular , Ratas , Ratas Endogámicas Lew , Ratas Endogámicas SHR , Ratas Endogámicas , Ratas WistarRESUMEN
Fifty-eight new anonymous simple sequence repeats (SSR) were generated and mapped to various rat chromosomes. Among them two genes (rat homologs for human cadherin-14 and mouse fibroblast growth factor-related protein) were mapped on Chromosomes (Chrs) 2 and 11 respectively. The majority of markers were generated from a small insert genomic library specific to Chr 11, 13, 14, and 15. Twenty new markers were mapped to Chr 13, which is known to contain a blood pressure quantitative trait locus (QTL). Several approaches to obtain microsatellite markers are described. The protocols and newly generated markers should be useful for ongoing rat genome project.
Asunto(s)
Mapeo Cromosómico/métodos , Ligamiento Genético , Repeticiones de Microsatélite , Ratas/genética , Animales , Secuencia de Bases , Cartilla de ADN/genética , Humanos , Ratones , Datos de Secuencia Molecular , Ratas EndogámicasRESUMEN
Human ATP1AL1 and corresponding genes of other mammals encode the catalytic alpha subunit of a non-gastric ouabain-sensitive H,K-ATPases, the ion pump presumably involved in maintenance of potassium homeostasis. The tissue specificity of the expression of these genes in different species has not been analyzed in detail. Here we report comparative RT-PCR screening of mouse, rat, rabbit, human, and dog tissues. Significant expression levels were observed in the skin, kidney and distal colon of all species (with the exception of the human colon). Analysis of rat urogenital organs also revealed strong expression in coagulating and preputial glands. Relatively lower expression levels were detected in many other tissues including brain, placenta and lung. In rabbit brain the expression was found to be specific to choroid plexus and cortex. Prominent similarity of tissue-specific expression patterns indicates that animal and human non-gastric H,K-ATPases are indeed products of homologous genes. This is also consistent with the high sequence similarity of non-gastric H,K-ATPases (including partial sequences of hitherto unknown cDNAs for mouse and dog proteins).
Asunto(s)
ATPasa Intercambiadora de Hidrógeno-Potásio/genética , Ouabaína/metabolismo , Secuencia de Aminoácidos , Animales , Encéfalo/enzimología , Dominio Catalítico , Colon/enzimología , Perros , Expresión Génica , ATPasa Intercambiadora de Hidrógeno-Potásio/metabolismo , Humanos , Riñón/enzimología , Ratones , Datos de Secuencia Molecular , Placenta/enzimología , Conejos , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Piel/enzimología , Distribución Tisular , Sistema Urogenital/enzimologíaRESUMEN
In studies designed to understand the roles of P2 nucleotide receptors in differentiation of T lymphocytes, we observed a transient and protein synthesis-independent enhancement of mRNA expression for the G protein-coupled P2Y2 receptor in mouse thymocytes after the addition of steroid hormone or T cell receptor (TCR) crosslinking by anti-TCR mAb. Conversely, dexamethasone-induced increases in mRNA expression for the ligand-gated ion channel P2X1 receptor was detected in rat, but not mouse, thymocytes, raising questions about the previously suggested role of P2X1 receptors in thymocyte apoptosis. Flow cytometry analysis of thymocyte subsets excluded the possibility that the observed increases in P2Y2 receptor mRNA expression were due to the enrichment of steroid-treated cells with an P2Y2 mRNA-rich thymocyte subset. Triggering of TCR-mediated intracellular signaling pathways through crosslinking of TCR or by addition of phorbol ester and Ca2+ ionophore also resulted in the up-regulation of P2Y2, but not P2X1, receptor mRNA. It is proposed that the rapid increase of P2Y2 receptor mRNA expression could be a common early event in responses of T cells to different activating stimuli. Taken together with the recently discovered ability of nucleotide receptor-initiated signaling to antagonize or enhance the effects of TCR crosslinking or steroids on thymocytes, the observed rapid up-regulation of P2Y2 receptor mRNA expression may reflect an immediate early gene response where newly expressed cell surface nucleotide receptors provide regulatory feedback signaling from extracellular ATP in the T cell differentiation process.
Asunto(s)
Regulación de la Expresión Génica/inmunología , Genes Inmediatos-Precoces/inmunología , Activación de Linfocitos/genética , Receptores Purinérgicos P2/genética , Linfocitos T/inmunología , Animales , Apoptosis , Células Cultivadas , Clonación Molecular , Cicloheximida/farmacología , Dexametasona/farmacología , Femenino , Glucocorticoides/farmacología , Humanos , Subgrupos Linfocitarios , Masculino , Ratones , Ratones Endogámicos DBA , Inhibidores de la Síntesis de la Proteína/farmacología , ARN Mensajero/biosíntesis , Receptores de Antígenos de Linfocitos T/fisiología , Receptores Purinérgicos P2X , Transducción de Señal , Linfocitos T/citología , Timo/citologíaRESUMEN
The human ATP1AL1 gene belongs to the family of Na,K-ATPase and H,K-ATPase (X,K-ATPases) genes. It encodes a catalytic subunit of hitherto unknown human ouabain-sensitive H,K-ATPase that represents a novel third group of X,K-ATPases distinct from the known Na,K-ATPase and gastric H,K-ATPase. Cloning of the ATP1AL1 gene is described in this report. The exon-intron structure of ATP1AL1 was found to be very similar to that of related genes. It contains 23 exons and spans approximately 32 kb of genomic DNA. All ATP1AL1 exons and 12 of its 22 introns were entirely sequenced. A total of nine Alu repeats were identified in introns. The transcription initiation site was mapped 187 bp upstream of the ATG initiation codon by primer extension and S1 nuclease protection analyses of RNA from human skin and colon. Sequence analysis of the 5'-flanking region (1.48 kb) revealed numerous potential binding sites for transcription factors Sp1 and AP2 and one putative NF-kappa B binding site. The 0.85-kb region from position -484 (5'-flanking region) to position +369 (intron 1) meets the structural criteria of a CpG island. It is suggested that the ATP1AL1 gene contains two poly(A) addition sites that may function in a tissue-specific manner.
Asunto(s)
Genes/genética , ATPasa Intercambiadora de Hidrógeno-Potásio/genética , ATPasa Intercambiadora de Sodio-Potasio/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Colon/química , Exones/genética , Humanos , Intrones/genética , Riñón/química , Datos de Secuencia Molecular , Ouabaína , ARN Mensajero/análisis , Secuencias Repetitivas de Ácidos Nucleicos/genética , Mapeo Restrictivo , Alineación de Secuencia , Análisis de Secuencia de ADN , Piel/química , Transcripción Genética/genéticaRESUMEN
The cDNA for ATP1AL1--the fifth member of the human Na,K-/H,K-ATPase gene family--was cloned and sequenced. The deduced primary ATP1AL1 translation product is 1,039 amino acids in length and has Mr of 114,543. The encoded protein has all of the structural features common to known catalytic subunits of P-type membrane ion-transporting ATPases and is equally distant (63-64% of identity) from the Na,K-ATPase isoforms and the gastric H,K-ATPase. The ATP1AL1 encoded protein was proposed to represent a new separate group within the family of human potassium-dependent ion pumps.
Asunto(s)
Adenosina Trifosfatasas/clasificación , ATPasa Intercambiadora de Hidrógeno-Potásio/genética , Familia de Multigenes/genética , ATPasa Intercambiadora de Sodio-Potasio/genética , Adenosina Trifosfatasas/genética , Secuencia de Aminoácidos , Secuencia de Bases , ADN Complementario/genética , ATPasa Intercambiadora de Hidrógeno-Potásio/química , ATPasa Intercambiadora de Hidrógeno-Potásio/clasificación , Humanos , Bombas Iónicas/metabolismo , Riñón/enzimología , Datos de Secuencia Molecular , Potasio/metabolismo , Homología de Secuencia de Aminoácido , Piel/enzimología , ATPasa Intercambiadora de Sodio-Potasio/química , ATPasa Intercambiadora de Sodio-Potasio/clasificación , Distribución TisularRESUMEN
The multigene family of human Na,K-ATPase is composed of 5 alpha-subunit genes, 3 of which were shown to encode the functionally active alpha 1, alpha 2 and alpha 3 isoforms of the catalytic subunits. This report describes the isolation, mapping and partial sequencing of the fourth gene (ATP1AL1) that was demonstrated here to be functionally active and expressed in human brain and kidney. Limited DNA sequencing of the ATP1AL1 exons allowed one to suggest that the gene probably encodes a new ion transport ATPase rather than an isoform of the Na,K-ATPase or the closely related H,K-ATPase.
Asunto(s)
Adenosina Trifosfatasas/genética , Familia de Multigenes , ATPasa Intercambiadora de Sodio-Potasio/genética , Transcripción Genética , Adenosina Trifosfatasas/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Transporte Biológico , Encéfalo/enzimología , Electroforesis en Gel de Agar , Exones , Humanos , Intrones , Riñón/enzimología , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Ácido NucleicoRESUMEN
Intra-individual tissue-specific restriction fragment length polymorphism (RFLP) has been demonstrated in DNA isolated from different mammalian tissues using cDNAs of alpha- and beta-subunits of Na+,K+-ATPase as hybridization probes. We propose that the RFLPs could result from gene rearrangements in the gene loci for the alpha- and beta-subunits of Na+,K+-ATPase. The changes in restriction patterns have been shown to occur during embryonic development and tumor formation. In addition, the tissue specificity of the expression of different genes of the family of Na+,K+-ATPase genes and their low expression in tumor cells have been demonstrated.
Asunto(s)
Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , ATPasa Intercambiadora de Sodio-Potasio/genética , Animales , ADN/genética , Feto/enzimología , Regulación de la Expresión Génica , Humanos , Ratones , Neoplasias/enzimología , Hibridación de Ácido Nucleico , ConejosRESUMEN
Among 356 gastric resections of the Billroth 11 type performed for ulcerous disease there were 11 cases of suture incompetency of the duodenal stump (3.1%). This complication was mainly caused by technical difficulties arising in operations under complicated anatomical conditions and tactical errors of a surgeon in selection of the technic of the stump suturing. The treatment of the duodenal stump suture incompetency consisted in urgent relaparotomy with a tramponade and drainage at the site of incompetency and application of a complex of conservative measures. During the first days after relaparotomy a drainage is connected into a pump-aspirator. In case of complicated duodenal ulcers resection with the aim of "ulcer exclusion" seems to be rational.
