20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua)
Abstract In order to better quantify the molecular mechanisms regulating final oocyte maturation and spawning, complete coding sequences with partially or fully untranslated regions for the steroidogenic enzymes, cytochrome P450 aromatase and 20β-hydroxysteroid dehydrogenase, were cloned from ovarie...
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crbioscientif:10.1677/jme-07-0070 2024-06-23T07:51:04+00:00 20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) Mittelholzer, C Andersson, E Consten, D Hirai, T Nagahama, Y Norberg, B 2007 http://dx.doi.org/10.1677/jme-07-0070 https://jme.bioscientifica.com/view/journals/jme/39/4/319.xml https://jme.bioscientifica.com/downloadpdf/journals/jme/39/4/319.xml unknown Bioscientifica Journal of Molecular Endocrinology volume 39, issue 4, page 319-328 ISSN 0952-5041 1479-6813 journal-article 2007 crbioscientif https://doi.org/10.1677/jme-07-0070 2024-06-06T04:11:08Z Abstract In order to better quantify the molecular mechanisms regulating final oocyte maturation and spawning, complete coding sequences with partially or fully untranslated regions for the steroidogenic enzymes, cytochrome P450 aromatase and 20β-hydroxysteroid dehydrogenase, were cloned from ovaries of Atlantic cod ( Gadus morhua ). The nucleotide and amino acid sequences showed high homologies with the corresponding sequences of other fish species, and conserved features important for functionality were identified in both predicted proteins. The sequences of the corresponding genomic loci were also determined, allowing the design of mRNA-specific quantitative PCR assays. As a reference gene for the real-time RT-PCR assays, eukaryotic elongation factor 1α was chosen, and the mRNA as well as the genomic sequence was determined. In addition, a real-time quantitative PCR assay for the 18S rRNA was adapted to be used in cod. Analysis of immature and maturing female cod from July to January respectively showed that the enzyme genes showed the expected quantitative changes associated with physiological regulation. However, mRNA for eukaryotic elongation factor 1α, and to a lesser extent even 18S rRNA, showed variable expression in these samples as well. To find accurate standards for real-time PCR in such a dynamic organ as the cod ovary is not an easy task, and several possible solutions are discussed. Article in Journal/Newspaper atlantic cod Gadus morhua Bioscientifica Journal of Molecular Endocrinology 39 4 319 328 |
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Abstract In order to better quantify the molecular mechanisms regulating final oocyte maturation and spawning, complete coding sequences with partially or fully untranslated regions for the steroidogenic enzymes, cytochrome P450 aromatase and 20β-hydroxysteroid dehydrogenase, were cloned from ovaries of Atlantic cod ( Gadus morhua ). The nucleotide and amino acid sequences showed high homologies with the corresponding sequences of other fish species, and conserved features important for functionality were identified in both predicted proteins. The sequences of the corresponding genomic loci were also determined, allowing the design of mRNA-specific quantitative PCR assays. As a reference gene for the real-time RT-PCR assays, eukaryotic elongation factor 1α was chosen, and the mRNA as well as the genomic sequence was determined. In addition, a real-time quantitative PCR assay for the 18S rRNA was adapted to be used in cod. Analysis of immature and maturing female cod from July to January respectively showed that the enzyme genes showed the expected quantitative changes associated with physiological regulation. However, mRNA for eukaryotic elongation factor 1α, and to a lesser extent even 18S rRNA, showed variable expression in these samples as well. To find accurate standards for real-time PCR in such a dynamic organ as the cod ovary is not an easy task, and several possible solutions are discussed. |
format |
Article in Journal/Newspaper |
author |
Mittelholzer, C Andersson, E Consten, D Hirai, T Nagahama, Y Norberg, B |
spellingShingle |
Mittelholzer, C Andersson, E Consten, D Hirai, T Nagahama, Y Norberg, B 20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
author_facet |
Mittelholzer, C Andersson, E Consten, D Hirai, T Nagahama, Y Norberg, B |
author_sort |
Mittelholzer, C |
title |
20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
title_short |
20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
title_full |
20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
title_fullStr |
20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
title_full_unstemmed |
20β-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua) |
title_sort |
20β-hydroxysteroid dehydrogenase and cyp19a1 are differentially expressed during maturation in atlantic cod (gadus morhua) |
publisher |
Bioscientifica |
publishDate |
2007 |
url |
http://dx.doi.org/10.1677/jme-07-0070 https://jme.bioscientifica.com/view/journals/jme/39/4/319.xml https://jme.bioscientifica.com/downloadpdf/journals/jme/39/4/319.xml |
genre |
atlantic cod Gadus morhua |
genre_facet |
atlantic cod Gadus morhua |
op_source |
Journal of Molecular Endocrinology volume 39, issue 4, page 319-328 ISSN 0952-5041 1479-6813 |
op_doi |
https://doi.org/10.1677/jme-07-0070 |
container_title |
Journal of Molecular Endocrinology |
container_volume |
39 |
container_issue |
4 |
container_start_page |
319 |
op_container_end_page |
328 |
_version_ |
1802642057653649408 |