Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin
Special issue Expanding the knowledge base of reproductive success: from genes to the environment.-- 11 pages, 7 figures In flatfishes with asynchronous and semicystic spermatogenesis, such as the Senegalese sole (Solea senegalensis), the specific roles of the pituitary gonadotropins during germ cel...
Published in: | General and Comparative Endocrinology |
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Academic Press
2015
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Online Access: | http://hdl.handle.net/10261/127392 https://doi.org/10.1016/j.ygcen.2014.10.009 https://doi.org/10.13039/100008054 https://doi.org/10.13039/501100004837 |
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ftcsic:oai:digital.csic.es:10261/127392 2024-02-11T10:09:17+01:00 Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin Chauvigné, François Verdura, Sara Mazón, María J. Boj, Mónica Gómez, Ana Cerdà, Joan Fundación Ramón Areces Ministerio de Ciencia e Innovación (España) 2015-09-15 http://hdl.handle.net/10261/127392 https://doi.org/10.1016/j.ygcen.2014.10.009 https://doi.org/10.13039/100008054 https://doi.org/10.13039/501100004837 unknown Academic Press http://dx.doi.org/10.1016/j.ygcen.2014.10.009 Sí doi:10.1016/j.ygcen.2014.10.009 issn: 0016-6480 e-issn: 1095-6840 General and Comparative Endocrinology 221: 75-85 (2015) http://hdl.handle.net/10261/127392 http://dx.doi.org/10.13039/100008054 http://dx.doi.org/10.13039/501100004837 none Flatfish Senegalese sole ELISA FSH Spermatogenesis artículo http://purl.org/coar/resource_type/c_6501 2015 ftcsic https://doi.org/10.1016/j.ygcen.2014.10.00910.13039/10000805410.13039/501100004837 2024-01-16T10:13:01Z Special issue Expanding the knowledge base of reproductive success: from genes to the environment.-- 11 pages, 7 figures In flatfishes with asynchronous and semicystic spermatogenesis, such as the Senegalese sole (Solea senegalensis), the specific roles of the pituitary gonadotropins during germ cell development, particularly of the follicle-stimulating hormone (Fsh), are still largely unknown in part due to the lack of homologous immunoassays for this hormone. In this study, an enzyme-linked immunosorbent assay (ELISA) for Senegalese sole Fsh was developed by generating a rabbit antiserum against a recombinant chimeric single-chain Fsh molecule (rFsh-C) produced by the yeast Pichia pastoris. The rFsh-C N- and C-termini were formed by the mature sole Fsh β subunit (Fshβ) and the chicken glycoprotein hormone common α subunit (CGA), respectively. Depletion of the antiserum to remove anti-CGA antibodies further enriched the sole Fshβ-specific antibodies, which were used to develop the ELISA using the rFsh-C for the standard curve. The sensitivity of the assay was 10 and 50 pg/ml for Fsh measurement in plasma and pituitary, respectively, and the cross-reactivity with a homologous recombinant single-chain luteinizing hormone was 1%. The standard curve for rFsh-C paralleled those of serially diluted plasma and pituitary extracts of other flatfishes, such as the Atlantic halibut, common sole and turbot. In Senegalese sole males, the highest plasma Fsh levels were found during early spermatogenesis but declined during enhanced spermiation, as found in teleosts with cystic spermatogenesis. In pubertal males, however, the circulating Fsh levels were as high as in adult spermiating fish, but interestingly the Fsh receptor in the developing testis containing only spermatogonia was expressed in Leydig cells but not in the primordial Sertoli cells. These results indicate that a recombinant chimeric Fsh can be used to generate specific antibodies against the Fshβ subunit and to develop a highly sensitive ELISA for Fsh ... Article in Journal/Newspaper Turbot Digital.CSIC (Spanish National Research Council) General and Comparative Endocrinology 221 75 85 |
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Digital.CSIC (Spanish National Research Council) |
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Flatfish Senegalese sole ELISA FSH Spermatogenesis |
spellingShingle |
Flatfish Senegalese sole ELISA FSH Spermatogenesis Chauvigné, François Verdura, Sara Mazón, María J. Boj, Mónica Gómez, Ana Cerdà, Joan Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
topic_facet |
Flatfish Senegalese sole ELISA FSH Spermatogenesis |
description |
Special issue Expanding the knowledge base of reproductive success: from genes to the environment.-- 11 pages, 7 figures In flatfishes with asynchronous and semicystic spermatogenesis, such as the Senegalese sole (Solea senegalensis), the specific roles of the pituitary gonadotropins during germ cell development, particularly of the follicle-stimulating hormone (Fsh), are still largely unknown in part due to the lack of homologous immunoassays for this hormone. In this study, an enzyme-linked immunosorbent assay (ELISA) for Senegalese sole Fsh was developed by generating a rabbit antiserum against a recombinant chimeric single-chain Fsh molecule (rFsh-C) produced by the yeast Pichia pastoris. The rFsh-C N- and C-termini were formed by the mature sole Fsh β subunit (Fshβ) and the chicken glycoprotein hormone common α subunit (CGA), respectively. Depletion of the antiserum to remove anti-CGA antibodies further enriched the sole Fshβ-specific antibodies, which were used to develop the ELISA using the rFsh-C for the standard curve. The sensitivity of the assay was 10 and 50 pg/ml for Fsh measurement in plasma and pituitary, respectively, and the cross-reactivity with a homologous recombinant single-chain luteinizing hormone was 1%. The standard curve for rFsh-C paralleled those of serially diluted plasma and pituitary extracts of other flatfishes, such as the Atlantic halibut, common sole and turbot. In Senegalese sole males, the highest plasma Fsh levels were found during early spermatogenesis but declined during enhanced spermiation, as found in teleosts with cystic spermatogenesis. In pubertal males, however, the circulating Fsh levels were as high as in adult spermiating fish, but interestingly the Fsh receptor in the developing testis containing only spermatogonia was expressed in Leydig cells but not in the primordial Sertoli cells. These results indicate that a recombinant chimeric Fsh can be used to generate specific antibodies against the Fshβ subunit and to develop a highly sensitive ELISA for Fsh ... |
author2 |
Fundación Ramón Areces Ministerio de Ciencia e Innovación (España) |
format |
Article in Journal/Newspaper |
author |
Chauvigné, François Verdura, Sara Mazón, María J. Boj, Mónica Gómez, Ana Cerdà, Joan |
author_facet |
Chauvigné, François Verdura, Sara Mazón, María J. Boj, Mónica Gómez, Ana Cerdà, Joan |
author_sort |
Chauvigné, François |
title |
Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
title_short |
Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
title_full |
Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
title_fullStr |
Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
title_full_unstemmed |
Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin |
title_sort |
development of a flatfish-specific enzyme-linked immunosorbent assay for fsh using a recombinant chimeric gonadotropin |
publisher |
Academic Press |
publishDate |
2015 |
url |
http://hdl.handle.net/10261/127392 https://doi.org/10.1016/j.ygcen.2014.10.009 https://doi.org/10.13039/100008054 https://doi.org/10.13039/501100004837 |
genre |
Turbot |
genre_facet |
Turbot |
op_relation |
http://dx.doi.org/10.1016/j.ygcen.2014.10.009 Sí doi:10.1016/j.ygcen.2014.10.009 issn: 0016-6480 e-issn: 1095-6840 General and Comparative Endocrinology 221: 75-85 (2015) http://hdl.handle.net/10261/127392 http://dx.doi.org/10.13039/100008054 http://dx.doi.org/10.13039/501100004837 |
op_rights |
none |
op_doi |
https://doi.org/10.1016/j.ygcen.2014.10.00910.13039/10000805410.13039/501100004837 |
container_title |
General and Comparative Endocrinology |
container_volume |
221 |
container_start_page |
75 |
op_container_end_page |
85 |
_version_ |
1790609102047543296 |