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14 results on '"Miliar, Angel"'

10. Natural trans-splicing in carnitine octanoyltransferase pre-mRNAs in rat liver

Author

Ministerio de Educación y Ciencia (España), Dirección General de Investigación Científica y Técnica, DGICT (España), Instituto de Cooperación Iberoamericana (España), Caudevilla, Concha, Serra, Dolors, Miliar, Angel, Codony, Carles, Asins, Guillermina, Bach-Elias, Montse, Hegardt, Fausto G., Ministerio de Educación y Ciencia (España), Dirección General de Investigación Científica y Técnica, DGICT (España), Instituto de Cooperación Iberoamericana (España), Caudevilla, Concha, Serra, Dolors, Miliar, Angel, Codony, Carles, Asins, Guillermina, Bach-Elias, Montse, and Hegardt, Fausto G.

Abstract

Carnitine octanoyltransferase (COT) transports medium-chain fatty acids through the peroxisome. During isolation of a COT clone from a rat liver library, a cDNA in which exon 2 was repeated, was characterized. Reverse transcription-PCR amplifications of total RNAs from rat liver showed a three-band pattern. Sequencing of the fragments revealed that, in addition to the canonical exon organization, previously reported [Choi, S. J. et al. (1995) Biochim. Biophys. Acta 1264, 215–222], there were two other forms in which exon 2 or exons 2 and 3 were repeated. The possibility of this exonic repetition in the COT gene was ruled out by genomic Southern blot. To study the gene expression, we analyzed RNA transcripts by Northern blot after RNase H digestion of total RNA. Three different transcripts were observed. Splicing experiments also were carried out in vitro with different constructs that contain exon 2 plus the 5′ or the 3′ adjacent intron sequences. Our results indicate that accurate joining of two exons 2 occurs by a trans-splicing mechanism, confirming the potential of these structures for this process in nature. The trans-splicing can be explained by the presence of three exon-enhancer sequences in exon 2. Analysis by Western blot of the COT proteins by using specific antibodies showed that two proteins corresponding to the expected Mr are present in rat peroxisomes. This is the first time that a natural trans-splicing reaction has been demonstrated in mammalian cells.

Published
1998

13. Differential expression of α1-adrenergic receptor subtypes in coronary microvascular endothelial cells in culture

Author

Mendez, Enrique, Calzada, Claudia, Ocharan, Esther, Sierra, Alfredo, Castillo, Carlos, Ramirez, Israel, Meaney, Eduardo, Meaney, Alejandra, Asbun, Juan, Miliar, Angel, Herrera, Jorge, and Ceballos, Guillermo

Subjects
*BLOOD vessels, *VASCULAR smooth muscle, *POLYMERASE chain reaction, *NITRIC oxide
Abstract

Abstract: It has been postulated that in blood vessels under α1-related stimulation, the endothelial intracellular calcium concentration ([Ca2+] i ) increases, which is necessary to induce nitric oxide synthesis, is the result of an increase in vascular smooth muscle, which subsequently, flows into the endothelial cells through gap junctions and it is not the result of a direct adrenergic stimulation of endothelial receptors. Others, however, postulate that endothelial α1D-adrenoceptors, have a direct effect on nitric oxide synthesis. In order to clarify this phenomena, in this work we analyzed the presence of α1receptor subtypes and their functional association with nitric oxide synthesis in rat coronary microvascular endothelial cells in culture, with pharmacological, immunological and reverse transcriptase polymerase chain reaction approaches. Our results show the presence and functional coupling with nitric oxide synthesis of α1A and α1D-adrenoceptor subtypes. α1B-adrenoceptor subtype is not coupled with nitric oxide production. [Copyright &y& Elsevier]

Published
2006
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14. Differential expression of alpha1-adrenergic receptor subtypes in coronary microvascular endothelial cells in culture.

Author

Mendez E, Calzada C, Ocharan E, Sierra A, Castillo C, Ramirez I, Meaney E, Meaney A, Asbun J, Miliar A, Herrera J, and Ceballos G

Subjects
Adrenergic alpha-Agonists pharmacology, Adrenergic alpha-Antagonists pharmacology, Animals, Bradykinin pharmacology, Cells, Cultured, Coronary Vessels cytology, Coronary Vessels drug effects, Coronary Vessels metabolism, Endothelial Cells drug effects, Endothelial Cells metabolism, Enzyme Inhibitors pharmacology, Fluorescent Antibody Technique, Indirect, Immunoblotting, Male, NG-Nitroarginine Methyl Ester pharmacology, Nitric Oxide biosynthesis, Nitric Oxide Synthase antagonists & inhibitors, Phenylephrine pharmacology, RNA, Messenger analysis, Rats, Rats, Wistar, Receptors, Adrenergic, alpha-1 drug effects, Reverse Transcriptase Polymerase Chain Reaction, Coronary Vessels chemistry, Endothelial Cells chemistry, Receptors, Adrenergic, alpha-1 analysis
Abstract

It has been postulated that in blood vessels under alpha(1)-related stimulation, the endothelial intracellular calcium concentration ([Ca(2+)](i)) increases, which is necessary to induce nitric oxide synthesis, is the result of an increase in vascular smooth muscle, which subsequently, flows into the endothelial cells through gap junctions and it is not the result of a direct adrenergic stimulation of endothelial receptors. Others, however, postulate that endothelial alpha(1D)-adrenoceptors, have a direct effect on nitric oxide synthesis. In order to clarify this phenomena, in this work we analyzed the presence of alpha(1)receptor subtypes and their functional association with nitric oxide synthesis in rat coronary microvascular endothelial cells in culture, with pharmacological, immunological and reverse transcriptase polymerase chain reaction approaches. Our results show the presence and functional coupling with nitric oxide synthesis of alpha(1A) and alpha(1D)-adrenoceptor subtypes. alpha(1B)-adrenoceptor subtype is not coupled with nitric oxide production.

Published
2006
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