Diseño y evaluación de primers para el estudio de la expresión del gen PepT1 en células del epitelio intestinal de juveniles de tilapia (Oreochromis sp.) por RT-PCR
Design and evaluation of primers for the study of gene expression of PepT1 in intestinal epithelial cells of juvenile tilapia (Oreochromis sp.) by RT-PCR
DOI:
https://doi.org/10.33936/la_tecnica.v0i13.587Palabras clave:
PepT1, SLC15A1, tilapias, Oreochromis niloticus, primers, RT-PCRResumen
El péptido transportador 1 (PepT1) es una proteína de membrana que facilita la captación de di/tri péptidos en el intestino. Estudios funcionales y estructurales han proporcionado información detallada sobre las características del gen PepT1 y varios aspectos de su regulación en vertebrados superiores, sin embargo, la información en vertebrados inferiores, donde se incluyen los peces, es limitada. PepT1 juega una importante función en la absorción de oligopéptidos y por lo tanto en el crecimiento de los peces. Los resultados de alineación de secuencias, usando el programa ClustalW, mostraron que PepT1 en tilapia (Oreochromis sp.) consiste de 21 exones, cubriendo una región genómica de unos 8 938 nucleótidos; mientras, la región codificante corresponde a 2 274 nucleótidos. Diferentes primers fueron diseñados basados en las regiones más homólogas de los alineamientos ClustalW. Seis pares de cebadores fueron seleccionados para el experimento de RT-PCR y el ARN extrayéndolo del intestino de juveniles de tilapia. Los amplicones esperados se obtuvieron por cada par de cebadores, en virtud de diversas temperaturas de hibridación y tiempo de amplificación.
Palabras clave: PepT1; SLC15A1; tilapia; Oreochromis niloticus; primers; RT-PCR.Descargas
La descarga de datos todavía no está disponible.
Citas
Amberg J. J., Myr C., Kamisaka Y., Jordal A.E.O., Rust M.B., Hardy R.W., Koedijk R., Ronnestad I. (2008). Expression of the oligopeptide transporter, PepT1, in larval Atlantic cod (Gadus morhua). Comp. Biochem. Physiol., B. 150, 177–182.
Bakke S., Jordal, A.E.O., Gómez-Requeni P., Verri, T., Kousoulaki K., Aksnes A., Ronnestad I. (2010). Dietary protein hydrolysates and free amino acids affect the spatial expression of peptide transporter PepT1 in the digestive tract of Atlantic cod (Gadus morhua). Comp. Biochem. and Physiol. B. 156, 48-55.
Boge, G., Rigal, A., Peres, G. (1981). Rates of in vivo absorption of glycine and glycineglycine by rainbow trout (Salmo gairdneri). Comp.
Biochem. Physiol. 69, 455–459.
Bucking C., & Schulte P. M. (2011).Environmental and nutritional regulation of expression and function of two peptide transporter (PepT1) isoforms in a euryhaline teleost. Comp. Biochem. and Physiol., A. 161, 379-387.
Dabrowski, K., Terjesen, B.F., Zhang, Y.F.,Phang, J.M., Lee, K.J. (2005). A concept of dietary dipeptides: a step to resolve the problem of amino acid availability in the early lifeof vertebrates. J. Exp. Biol. 208, 2885–2894.
Dabrowski, K., Lee, K.J., Rinchard, J. (2003).The smallest vertebrate, teleost fish, can utilize synthetic dipeptide-based diets. J. Nutr. 133,4225–4229.
Daniel, H. (2004). Molecular and integrative physiology of intestinal peptide transport. Annu. Rev. Physiol. 66, 361–384.
Daniel, H., Kottra, G. (2004). The proton oligopeptide cotransporter family SLC15 inphysiology and pharmacology. Pfluegers Arch. Eur. J. Physiol. 447, 610–618.
Gilbert, E. R., Li, H., Emmerson, D. A.,Webb Jr, K. E. & Wong, E. A. (2008). Dietary protein quality and feed restriction influence abundance of nutrient transporter mRNA in the small intestine of broiler chicks. J. Nutr. 138,262–271.
Gilbert, E.R., Wong, E.A., Webb Jr., K.E. (2008b). Board-invited review: Peptide absorption and utilization: Implications for animal nutrition and health. J. Anim. Sci. 86,2135–2155.
Hakim, Y., Harpaz, S., Uni, Z. (2009). Expression of brush border enzymes and transporter in the intestine of European sea
bass (Dicentrarchus labrax) following food deprivation. Aquacultural 290, 110–115.
Ihara, T., Tsujikawa, T., Fujiyama, Y., Bamba,T.(2000). Regulation of PepT1 peptide transporter expression in the rat small intestine under malnourished conditions. Digestion 61,59–67.
Kalujnaia, S., McWilliam, I.S., Zaguinaiko,V.A., Feilen, A.L., Nicholson, J., Hazon, N., Cutler, C.P., Cramb, G. (2007). Transcriptomic approach to the study of osmoregulation in the European el Anguilla anguilla. Physiol. Genomics. 31, 385–401.
Leibach, F.H., Ganapathy, V. (1996). Peptide transporters of the intestine and kidney.Annu.Rev. Nutr. 16, 99–119.
Meredith, D., Boyd, C.A.R. (2000). Structure and function of eukaryotic peptide transporters. Cell. Mol. Life Sci. 57, 754–778.
Ogihara, H., Suzuki, T., Nagamachi, Y., Inui, K. & Takata, K. (1999). Peptide transporter in the rat small intestine: ultrastructural localization
and the effect of starvation and administration of amino acids. Histochem. J. 31, 169–174.
Ostaszewska, T., Kamaszewski, M.,Grochowski, P., Dabrowski, K., Verri, T., Aksakal, E., Szatkowska, I., Nowak, Z., Dobosz, S. (2010). The effect of peptide absorption on PepT1 gene expression and digestive system hormones in rainbow trout (Oncorhynchus mykiss). Comp. Biochem.Physiol. 155, 107–114.
Reshkin, S.J., Ahearn, G.A. (1991). Intestinal glycyl-L-phenylalanine and L-phenylalanine transport in a euryhaline teleost. Am. J. Physiol. 260, R563–R569.
Ronnestad, I., Murashita, K., Kottra, G., Jordal, A.E., Narawane, S., Jolly, C., Daniel,H., Verri, T. (2010). Molecular cloning and functional expression of Atlantic salmon peptide transporter 1 in xenopus oocytes reveals efficient intestinal uptake of lysine-containing and other bioactive di- and tripeptides in teleost
fish. J. Nutr. 140, 893–900.
Ronnestad, I., Gavaia, P.J., Viegas, C.S.B.,Verri, T., Romano, A., Nilsen, T.O., Jordal,A.-E.O.,Kamisaka, Y., Cancela, M.L. (2007b). Cloning, tissue expression and comparative aspects of the Atlantic cod (Gadus morhua L.) oligopeptide transporter PepT1. J. Exp. Biol. 210, 3883–3896.
Ronnestad I., Gavaia P.J., Viegas C.S.B., Verri T., Romano A., Nilsen T.O., Jordal A.E., Kamisaka Y., Cancela M.L. (2007).Oligopeptide transporter PepT1 in Atlantic cod (Gadus morhua L.): cloning, tissue expression and comparative aspects. J. Exp. Biol. 210,3883–3896.
Rubio-Aliaga, I., Daniel, H. (2002). Mammalian peptide transporters as targets for drug delivery.
Trends Pharmacol. Sci. 23, 434–440.
Sangaletti, R., Terova, G., Peres, A., Bossi,E., Cora, S., Saroglia, M. (2009). Functional expression of the oligopeptide transporter PepT1 from the sea bass (Dicentrarchus labrax). Pflugers Arch. Eur. J. Physiol. 459, 47–54.
Segura, M., Chel, L., Betancur, D. (2010).Efecto de la digestión en la biodisponibilidad de péptidos con actividad biológica. Rev Chil Nutr Vol. 37, N°3, 386-391.
Terova, G., Cora, S., Verri,T., Rimoldi, S.,Bernardini, G. & Saroglia, M. (2009). Impact of feed availability on PEPT1 mRNA expression levels in sea bass (Dicentrarchus labrax).Aquaculture. 294, 288–299.
Thamotharan, M., Bawani, S.Z., Zhou, X.D.,Adibi, S.A. (1999). Functional and molecular expression of intestinal oligopeptide transporter (Pept-1) after a brief fast. Metabol. Clin. Exp.48, 681–684.
Thwaites, D.T., Anderson, C.M.H. (2007).H+-coupled nutrient, micronutrient and drug transporters in the mammalian small intestine. Exp. Physiol. 92, 603–619.
Verri, T., Terova, G., Dabrowski, K., and Saroglia, M. (2011). Peptide transport and animal growth: the fish paradigm. Biol. Lett. 7, 597-600.
Verri, T., Kottra, G., Romano, A., Tiso, N., Peric, M., Maffia, M., Boll, M., Argenton, F., Daniel, H., Storelli, C. (2003). Molecular and functional characterization of the zebrafish (Danio rerio) PepT1-type peptide transporter.FEBS Lett. 549, 115-122.
Bakke S., Jordal, A.E.O., Gómez-Requeni P., Verri, T., Kousoulaki K., Aksnes A., Ronnestad I. (2010). Dietary protein hydrolysates and free amino acids affect the spatial expression of peptide transporter PepT1 in the digestive tract of Atlantic cod (Gadus morhua). Comp. Biochem. and Physiol. B. 156, 48-55.
Boge, G., Rigal, A., Peres, G. (1981). Rates of in vivo absorption of glycine and glycineglycine by rainbow trout (Salmo gairdneri). Comp.
Biochem. Physiol. 69, 455–459.
Bucking C., & Schulte P. M. (2011).Environmental and nutritional regulation of expression and function of two peptide transporter (PepT1) isoforms in a euryhaline teleost. Comp. Biochem. and Physiol., A. 161, 379-387.
Dabrowski, K., Terjesen, B.F., Zhang, Y.F.,Phang, J.M., Lee, K.J. (2005). A concept of dietary dipeptides: a step to resolve the problem of amino acid availability in the early lifeof vertebrates. J. Exp. Biol. 208, 2885–2894.
Dabrowski, K., Lee, K.J., Rinchard, J. (2003).The smallest vertebrate, teleost fish, can utilize synthetic dipeptide-based diets. J. Nutr. 133,4225–4229.
Daniel, H. (2004). Molecular and integrative physiology of intestinal peptide transport. Annu. Rev. Physiol. 66, 361–384.
Daniel, H., Kottra, G. (2004). The proton oligopeptide cotransporter family SLC15 inphysiology and pharmacology. Pfluegers Arch. Eur. J. Physiol. 447, 610–618.
Gilbert, E. R., Li, H., Emmerson, D. A.,Webb Jr, K. E. & Wong, E. A. (2008). Dietary protein quality and feed restriction influence abundance of nutrient transporter mRNA in the small intestine of broiler chicks. J. Nutr. 138,262–271.
Gilbert, E.R., Wong, E.A., Webb Jr., K.E. (2008b). Board-invited review: Peptide absorption and utilization: Implications for animal nutrition and health. J. Anim. Sci. 86,2135–2155.
Hakim, Y., Harpaz, S., Uni, Z. (2009). Expression of brush border enzymes and transporter in the intestine of European sea
bass (Dicentrarchus labrax) following food deprivation. Aquacultural 290, 110–115.
Ihara, T., Tsujikawa, T., Fujiyama, Y., Bamba,T.(2000). Regulation of PepT1 peptide transporter expression in the rat small intestine under malnourished conditions. Digestion 61,59–67.
Kalujnaia, S., McWilliam, I.S., Zaguinaiko,V.A., Feilen, A.L., Nicholson, J., Hazon, N., Cutler, C.P., Cramb, G. (2007). Transcriptomic approach to the study of osmoregulation in the European el Anguilla anguilla. Physiol. Genomics. 31, 385–401.
Leibach, F.H., Ganapathy, V. (1996). Peptide transporters of the intestine and kidney.Annu.Rev. Nutr. 16, 99–119.
Meredith, D., Boyd, C.A.R. (2000). Structure and function of eukaryotic peptide transporters. Cell. Mol. Life Sci. 57, 754–778.
Ogihara, H., Suzuki, T., Nagamachi, Y., Inui, K. & Takata, K. (1999). Peptide transporter in the rat small intestine: ultrastructural localization
and the effect of starvation and administration of amino acids. Histochem. J. 31, 169–174.
Ostaszewska, T., Kamaszewski, M.,Grochowski, P., Dabrowski, K., Verri, T., Aksakal, E., Szatkowska, I., Nowak, Z., Dobosz, S. (2010). The effect of peptide absorption on PepT1 gene expression and digestive system hormones in rainbow trout (Oncorhynchus mykiss). Comp. Biochem.Physiol. 155, 107–114.
Reshkin, S.J., Ahearn, G.A. (1991). Intestinal glycyl-L-phenylalanine and L-phenylalanine transport in a euryhaline teleost. Am. J. Physiol. 260, R563–R569.
Ronnestad, I., Murashita, K., Kottra, G., Jordal, A.E., Narawane, S., Jolly, C., Daniel,H., Verri, T. (2010). Molecular cloning and functional expression of Atlantic salmon peptide transporter 1 in xenopus oocytes reveals efficient intestinal uptake of lysine-containing and other bioactive di- and tripeptides in teleost
fish. J. Nutr. 140, 893–900.
Ronnestad, I., Gavaia, P.J., Viegas, C.S.B.,Verri, T., Romano, A., Nilsen, T.O., Jordal,A.-E.O.,Kamisaka, Y., Cancela, M.L. (2007b). Cloning, tissue expression and comparative aspects of the Atlantic cod (Gadus morhua L.) oligopeptide transporter PepT1. J. Exp. Biol. 210, 3883–3896.
Ronnestad I., Gavaia P.J., Viegas C.S.B., Verri T., Romano A., Nilsen T.O., Jordal A.E., Kamisaka Y., Cancela M.L. (2007).Oligopeptide transporter PepT1 in Atlantic cod (Gadus morhua L.): cloning, tissue expression and comparative aspects. J. Exp. Biol. 210,3883–3896.
Rubio-Aliaga, I., Daniel, H. (2002). Mammalian peptide transporters as targets for drug delivery.
Trends Pharmacol. Sci. 23, 434–440.
Sangaletti, R., Terova, G., Peres, A., Bossi,E., Cora, S., Saroglia, M. (2009). Functional expression of the oligopeptide transporter PepT1 from the sea bass (Dicentrarchus labrax). Pflugers Arch. Eur. J. Physiol. 459, 47–54.
Segura, M., Chel, L., Betancur, D. (2010).Efecto de la digestión en la biodisponibilidad de péptidos con actividad biológica. Rev Chil Nutr Vol. 37, N°3, 386-391.
Terova, G., Cora, S., Verri,T., Rimoldi, S.,Bernardini, G. & Saroglia, M. (2009). Impact of feed availability on PEPT1 mRNA expression levels in sea bass (Dicentrarchus labrax).Aquaculture. 294, 288–299.
Thamotharan, M., Bawani, S.Z., Zhou, X.D.,Adibi, S.A. (1999). Functional and molecular expression of intestinal oligopeptide transporter (Pept-1) after a brief fast. Metabol. Clin. Exp.48, 681–684.
Thwaites, D.T., Anderson, C.M.H. (2007).H+-coupled nutrient, micronutrient and drug transporters in the mammalian small intestine. Exp. Physiol. 92, 603–619.
Verri, T., Terova, G., Dabrowski, K., and Saroglia, M. (2011). Peptide transport and animal growth: the fish paradigm. Biol. Lett. 7, 597-600.
Verri, T., Kottra, G., Romano, A., Tiso, N., Peric, M., Maffia, M., Boll, M., Argenton, F., Daniel, H., Storelli, C. (2003). Molecular and functional characterization of the zebrafish (Danio rerio) PepT1-type peptide transporter.FEBS Lett. 549, 115-122.
Publicado
2014-12-01
Cómo citar
Vera Delgado, J. M. (2014). Diseño y evaluación de primers para el estudio de la expresión del gen PepT1 en células del epitelio intestinal de juveniles de tilapia (Oreochromis sp.) por RT-PCR: Design and evaluation of primers for the study of gene expression of PepT1 in intestinal epithelial cells of juvenile tilapia (Oreochromis sp.) by RT-PCR. La Técnica. Revista De Las Agrociencias. ISSN 2477-8982, (13), 54–63. https://doi.org/10.33936/la_tecnica.v0i13.587
Número
Sección
Ciencias de la Vida
