RESUMEN
PURPOSE: To explore the effect of docosahexaenoic acid (DHA) on gene expression during human fetal retinal maturation. METHODS: Human fetal retinal explants were cultured in serum-free Waymouth's medium supplemented with DHA or oleic acid (OA), using bovine serum albumin (BSA) as the vehicle. After 14 days in culture, fatty acid composition was assessed, and the abundance of 2400 cDNAs was examined with a human cDNA microarray system. RESULTS: Transcript abundance remained unchanged for 82% and 90% of genes in the explants with added DHA or OA, respectively. Decreased expression was detected in 4% and 9% of genes, in explants supplemented with DHA or OA, respectively, whereas, 14% of genes in explants exposed to DHA and only 0.4% of genes in explants treated with OA showed increased expression. Transcripts displaying changes in abundance in explants supplemented with DHA encode for proteins involved in diverse biological functions, including neurogenesis, neurotransmission, and refinement of connectivity. These gene expression changes were not observed in explants supplemented with OA. CONCLUSIONS: The effect of DHA deficiency on retinal function during human development can be partly explained by modifications in retinal gene expression by direct or indirect mechanisms.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Retina/efectos de los fármacos , ADN Complementario/análisis , Ácidos Grasos/metabolismo , Feto , Perfilación de la Expresión Génica , Humanos , Metabolismo de los Lípidos , Análisis de Secuencia por Matrices de Oligonucleótidos , Técnicas de Cultivo de Órganos , Retina/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Peroxisomal proliferator-activated receptors (PPAR) are a FA-response system involved in diverse cellular responses. FA regulate PPAR activity and modulate PPAR mRNA abundance. Increasing evidence indicates that PUFA are required for optimal neuronal development and function. To gain insight into the mechanism for nutrition-induced impairment of neuronal development and function we investigated the effect of chronic n-3 FA deficiency on PPAR mRNA levels in rat brain and ocular tissues. Rats were fed for three generations a diet designed to reduce DHA levels in tissues, and the abundance of PPARalpha and PPARbeta transcripts was measured by hybridization with specific probes. Chronic consumption of the a-linolenic acid (LNA)-insufficient diet caused a remarkable modification in DHA content in membrane phospholipids. The results reported here indicate that PPARa mRNA levels did not exhibit significant variation in ocular, hepatic, or nervous tissues from rats fed the experimental diet. In contrast, PPARalpha mRNA normalized to beta-actin mRNA was 21% higher in ocular tissue from F3 generation rats consuming the LNA-deficient diet but was independent of diet in hepatic and nervous tissues. The absolute abundance of PPARbeta transcripts showed a 17% increase in ocular tissue from rats consuming the LNA-deficient diet (F3 generation). The biological significance of the reported changes in PPARbeta mRNA in ocular tissue remains to be determined.