Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay

 The luteinizing hormone (LH) is a key regulator of the processes of gonad maturation, ovulation/spermiation and spawning in vertebrates. The present study describes the purification of LH from pituitaries of a teleost fish, the European sea bass (Dicentrarchus labrax), by three-step chroma...

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Autores principales:	Mateos, J, Mañanós, E, Swanson, P, Carrillo, M, Zanuy, S
Formato:	info:eu-repo/semantics/article
Lenguaje:	eng
Publicado:	Iniversidad Autónoma de Baja California 2006
Materias:	reproduction luteinizing hormone immunoassay sea bass fish reproducción hormona luteinizante inmunoensayo lubina peces
Acceso en línea:	https://www.cienciasmarinas.com.mx/index.php/cmarinas/article/view/1062
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	repositorioinstitucional-20.500.12930-7401
record_format	dspace
institution	Repositorio Institucional
collection	DSpace
language	eng
topic	reproduction luteinizing hormone immunoassay sea bass fish reproducción hormona luteinizante inmunoensayo lubina peces
spellingShingle	reproduction luteinizing hormone immunoassay sea bass fish reproducción hormona luteinizante inmunoensayo lubina peces Mateos, J Mañanós, E Swanson, P Carrillo, M Zanuy, S Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
description	The luteinizing hormone (LH) is a key regulator of the processes of gonad maturation, ovulation/spermiation and spawning in vertebrates. The present study describes the purification of LH from pituitaries of a teleost fish, the European sea bass (Dicentrarchus labrax), by three-step chromatography (gel filtration, ion-exchange and FPLC). The LH α and β subunits were isolated by rpHPLC. The molecular weight of LH was estimated, on SDS-PAGE, at 31 kD, and for its α and β subunits, at 12 and 22 kD, respectively. Specific antibodies against the sea bass LHβ subunit (AbLHβ obtained and used to develop a specific enzyme-linked immunosorbent assay (ELISA), which had a sensitivity of around 0.65 ng mL–1 (Bi/Bo 80%) and intraand inter-assay coefficients of variation of 11.7% (n = 8) and 11% (n = 10), respectively. Validation of the assay showed parallelism between the standard curve and plasma and pituitary samples from sea bass, as well as with pituitary extracts from other perciform fishes. We measured, using ELISA, plasma LH levels in female sea bass given a single injection of different doses of GnRHa ([D-Ala6, Pro9-Net]-LHRH), as a hormonal therapy for spawning induction. All treatments increased plasma LH levels after 90 min of the injection and were maintained elevated during 24 h. In conclusion, the immunoassay developed is sensitive and accurate, useful for LH analysis in biological samples of sea bass, and represents a valuable tool for studies on the reproductive endocrinology of this species.
format	info:eu-repo/semantics/article
author	Mateos, J Mañanós, E Swanson, P Carrillo, M Zanuy, S
author_facet	Mateos, J Mañanós, E Swanson, P Carrillo, M Zanuy, S
author_sort	Mateos, J
title	Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
title_short	Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
title_full	Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
title_fullStr	Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
title_full_unstemmed	Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay
title_sort	purification of luteinizing hormone (lh) in the sea bass (dicentrarchus labrax) and development of a specific immunoassay
publisher	Iniversidad Autónoma de Baja California
publishDate	2006
url	https://www.cienciasmarinas.com.mx/index.php/cmarinas/article/view/1062
_version_	1792610797510197248
spelling	repositorioinstitucional-20.500.12930-74012023-05-09T14:30:59Z Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay Purificación de la hormona luteinizante (LH) en la lubina (Dicentrarchus labrax) y desarrollo de un inmunoensayo específico Mateos, J Mañanós, E Swanson, P Carrillo, M Zanuy, S reproduction luteinizing hormone immunoassay sea bass fish reproducción hormona luteinizante inmunoensayo lubina peces  The luteinizing hormone (LH) is a key regulator of the processes of gonad maturation, ovulation/spermiation and spawning in vertebrates. The present study describes the purification of LH from pituitaries of a teleost fish, the European sea bass (Dicentrarchus labrax), by three-step chromatography (gel filtration, ion-exchange and FPLC). The LH α and β subunits were isolated by rpHPLC. The molecular weight of LH was estimated, on SDS-PAGE, at 31 kD, and for its α and β subunits, at 12 and 22 kD, respectively. Specific antibodies against the sea bass LHβ subunit (AbLHβ obtained and used to develop a specific enzyme-linked immunosorbent assay (ELISA), which had a sensitivity of around 0.65 ng mL–1 (Bi/Bo 80%) and intraand inter-assay coefficients of variation of 11.7% (n = 8) and 11% (n = 10), respectively. Validation of the assay showed parallelism between the standard curve and plasma and pituitary samples from sea bass, as well as with pituitary extracts from other perciform fishes. We measured, using ELISA, plasma LH levels in female sea bass given a single injection of different doses of GnRHa ([D-Ala6, Pro9-Net]-LHRH), as a hormonal therapy for spawning induction. All treatments increased plasma LH levels after 90 min of the injection and were maintained elevated during 24 h. In conclusion, the immunoassay developed is sensitive and accurate, useful for LH analysis in biological samples of sea bass, and represents a valuable tool for studies on the reproductive endocrinology of this species. La hormona luteinizante (LH), secretada por la glándula hipofisaria, regula los procesos de maduración gonadal, ovulación/ espermiación y puesta en vertebrados. El presente trabajo describe la purificación de la LH de un pez teleósteo, la lubina europea (Dicentrarchus labrax), mediante triple cromatografía en columna (gel filtración, intercambio iónico y FPLC). Las subunidades α y β de la LH se aislaron mediante rpHPLC. El peso molecular de la LH se estimó, mediante SDS-PAGE, en 31 kD y el de sus subunidades α y β en 12 y 22 kD, respectivamente. Se obtuvieron anticuerpos específicos contra la subunidad LHβ (AbLHβ), que fueron utilizados para desarrollar un inmunoensayo tipo ELISA. La sensibilidad del ELISA fue de unos 0.65 ng mL–1 (Bi/Bo 80%) y los coeficientes de variación intra e interensayo, de 11.7% (n = 8) y 11% (n = 10), respectivamente. La validación del ensayo mostró paralelismo entre la curva patrón (LH) y muestras de plasma e hipófisis de lubina, así como con extractos hipofisiarios de otras especies de peces perciformes. Mediante ELISA, se analizaron los niveles plasmáticos de LH en lubinas sometidas a tratamiento de inducción hormonal a la puesta, por inyección simple de diferentes dosis de GnRHa ([D-Ala6, Pro9- Net]-LHRH). Todos los tratamientos provocaron un marcado incremento de la LH plasmática a los 90 min después de la inyección, manteniéndose elevados durante 24 h. En conclusión, el ELISA puesto a punto es un inmunoensayo sensible y preciso, apropiado para cuantificar LH en muestras biológicas de lubina y representa una valiosa herramienta para realizar estudios sobre la endocrinología de la reproducción de esta especie. 2006-06-23 2021-06-03T03:55:11Z 2021-06-03T03:55:11Z info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Peer-reviewed Article Artículo Arbitrado https://www.cienciasmarinas.com.mx/index.php/cmarinas/article/view/1062 10.7773/cm.v32i21.1062 https://hdl.handle.net/20.500.12930/7401 eng https://www.cienciasmarinas.com.mx/index.php/cmarinas/article/view/1062/946 application/pdf Iniversidad Autónoma de Baja California Ciencias Marinas; Vol. 32 No. 2 (2006); 271-283 Ciencias Marinas; Vol. 32 Núm. 2 (2006); 271-283 2395-9053 0185-3880

Purification of luteinizing hormone (LH) in the sea bass (Dicentrarchus labrax) and development of a specific immunoassay

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