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Int J Parasitol ; 38(2): 191-202, 2008 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17803998

RESUMEN

Caveolins are integral membrane proteins implicated in cholesterol homeostasis and transport, endocytosis mechanisms and regulation of signal transduction in differentiated cells. In this work a caveolin-1 gene from the nematode Trichinella spiralis (Ts-cav-1) was cloned and identified as an adult-specific antigen. For this, a cDNA library of T. spiralis 3-day-old adult worms was screened using a stage-specific cDNA-labelled probe. One positive clone contained a cDNA insert of 1427-bp and a full-length open reading frame (ORF) of 687-bp, which encodes for a 229 amino acid polypeptide with a theoretical molecular weight of 26kDa. BLAST and FASTA searches revealed a 36% and 57% identity with Caenorhabditis elegans caveolin-1, respectively. Confocal laser microscopy analysis using antibodies generated against Ts-CAV-1 protein and cross-sections of adult parasites showed that Ts-CAV-1 gradually accumulates on the surface of Trichinella oocytes and embryos, reaching a maximum at 3days p.i., and decreasing during new-born larvae (NBL) development. RT-PCR assays of parasites from 1 to 4days p.i. showed a similar gene expression profile to that observed for Ts-CAV-1 which suggests a specific developmental regulation. Free cholesterol was mainly distributed in the female germ line and it displayed increasing membrane accumulation, similar to the pattern obtained for Ts-CAV-1 protein, which suggests a temporal membrane association with Ts-CAV-1 that in turn will perform the functions mentioned above. Our results strongly indicate that Ts-cav-1 from T. spiralis plays a role in oocyte maturation and embryogenesis during development, demonstrating gender-specific expression.


Asunto(s)
Caveolina 1/aislamiento & purificación , Embrión no Mamífero/metabolismo , Regulación del Desarrollo de la Expresión Génica , Oocistos/metabolismo , Trichinella spiralis/embriología , Trichinella spiralis/metabolismo , Secuencia de Aminoácidos , Animales , Antígenos Helmínticos/genética , Secuencia de Bases , Western Blotting/métodos , Caveolina 1/genética , Caveolina 1/metabolismo , Caveolina 3/genética , Femenino , Expresión Génica , Microscopía Confocal , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos
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