Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development
We analyzed the presence of 3β-Hydroxysteroid Dehydrogenase/Δ5→4-isomerase enzyme (3β-HSD) activity, a key enzyme of the steroid metabolic pathway, the mRNA of this enzyme, and the steroid metabolism in in vitro produced bovine embryos. 3β-HSD activity was detected in in vitro matured oocytes (74.4±...
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todo:paper_1040452X_v61_n2_p135_Chiappe2023-10-03T15:57:50Z Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development Chiappe, M.E. Lattanzi, M.L. Colman-Lerner, A.A. Barañao, J.L. Saragüeta, P. Bovine Embryo development Gene expression Steroids 3(or 17)beta hydroxysteroid dehydrogenase alpha amanitin aromatase inhibitor transcription factor animal tissue article blastocyst cattle embryo development enzyme activity gene expression regulation messenger RNA synthesis nonhuman phase transition pregnancy preimplantation embryo priority journal reverse transcription polymerase chain reaction steroid metabolism steroidogenesis 3-Hydroxysteroid Dehydrogenases Amanitins Animals Blastocyst Cattle Cells, Cultured Dehydroepiandrosterone Embryonic and Fetal Development Female Fertilization in Vitro Nucleic Acid Synthesis Inhibitors Oocytes Pregnancy Pregnenolone Progesterone RNA, Messenger Bovinae We analyzed the presence of 3β-Hydroxysteroid Dehydrogenase/Δ5→4-isomerase enzyme (3β-HSD) activity, a key enzyme of the steroid metabolic pathway, the mRNA of this enzyme, and the steroid metabolism in in vitro produced bovine embryos. 3β-HSD activity was detected in in vitro matured oocytes (74.4±1.4%), 1-cell (72.9±6.1%), 2-cell (61.8±7.4%), 8-cell (50±5%), morulae (50.8±2.6%), blastocysts (94.4±3%), and hatched blastocysts (100±0%) meanwhile the 4-cell stage showed a significant reduction (16.7±4.7%). When total embryonic RNA of different stages was subjected to RT-PCR assays, the mRNA of 3β-HSD was found to be present in all developmental stages of in vitro produced bovine embryos, from the oocyte to the blastocyst, with a marked decrease at the 4-cell stage. To determine whether the temporal pattern of enzyme activity was dependent on the maternal to zygotic transition, embryos were incubated in the presence of a transcription inhibitor, α-amanitin. The reappearance of the enzyme activity after the 4-cell stage was blocked in α-amanitin treated embryos, indicating the requirement of embryonic transcription. On the other hand, the embryonic steroid metabolism was tested by incubating blastocyst with tritiated pregnenolone. Analysis of the metabolites by TLC indicated the production of a compound with a mobility identical to progesterone. These results described the expression of the 3β-HSD and the activity of this metabolic enzyme in bovine oocytes and preimplantation embryos, suggesting that steroids may act as autocrine effectors on preimplantation embryo development. © 2002 Wiley-Liss, Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_1040452X_v61_n2_p135_Chiappe |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Bovine Embryo development Gene expression Steroids 3(or 17)beta hydroxysteroid dehydrogenase alpha amanitin aromatase inhibitor transcription factor animal tissue article blastocyst cattle embryo development enzyme activity gene expression regulation messenger RNA synthesis nonhuman phase transition pregnancy preimplantation embryo priority journal reverse transcription polymerase chain reaction steroid metabolism steroidogenesis 3-Hydroxysteroid Dehydrogenases Amanitins Animals Blastocyst Cattle Cells, Cultured Dehydroepiandrosterone Embryonic and Fetal Development Female Fertilization in Vitro Nucleic Acid Synthesis Inhibitors Oocytes Pregnancy Pregnenolone Progesterone RNA, Messenger Bovinae |
| spellingShingle |
Bovine Embryo development Gene expression Steroids 3(or 17)beta hydroxysteroid dehydrogenase alpha amanitin aromatase inhibitor transcription factor animal tissue article blastocyst cattle embryo development enzyme activity gene expression regulation messenger RNA synthesis nonhuman phase transition pregnancy preimplantation embryo priority journal reverse transcription polymerase chain reaction steroid metabolism steroidogenesis 3-Hydroxysteroid Dehydrogenases Amanitins Animals Blastocyst Cattle Cells, Cultured Dehydroepiandrosterone Embryonic and Fetal Development Female Fertilization in Vitro Nucleic Acid Synthesis Inhibitors Oocytes Pregnancy Pregnenolone Progesterone RNA, Messenger Bovinae Chiappe, M.E. Lattanzi, M.L. Colman-Lerner, A.A. Barañao, J.L. Saragüeta, P. Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| topic_facet |
Bovine Embryo development Gene expression Steroids 3(or 17)beta hydroxysteroid dehydrogenase alpha amanitin aromatase inhibitor transcription factor animal tissue article blastocyst cattle embryo development enzyme activity gene expression regulation messenger RNA synthesis nonhuman phase transition pregnancy preimplantation embryo priority journal reverse transcription polymerase chain reaction steroid metabolism steroidogenesis 3-Hydroxysteroid Dehydrogenases Amanitins Animals Blastocyst Cattle Cells, Cultured Dehydroepiandrosterone Embryonic and Fetal Development Female Fertilization in Vitro Nucleic Acid Synthesis Inhibitors Oocytes Pregnancy Pregnenolone Progesterone RNA, Messenger Bovinae |
| description |
We analyzed the presence of 3β-Hydroxysteroid Dehydrogenase/Δ5→4-isomerase enzyme (3β-HSD) activity, a key enzyme of the steroid metabolic pathway, the mRNA of this enzyme, and the steroid metabolism in in vitro produced bovine embryos. 3β-HSD activity was detected in in vitro matured oocytes (74.4±1.4%), 1-cell (72.9±6.1%), 2-cell (61.8±7.4%), 8-cell (50±5%), morulae (50.8±2.6%), blastocysts (94.4±3%), and hatched blastocysts (100±0%) meanwhile the 4-cell stage showed a significant reduction (16.7±4.7%). When total embryonic RNA of different stages was subjected to RT-PCR assays, the mRNA of 3β-HSD was found to be present in all developmental stages of in vitro produced bovine embryos, from the oocyte to the blastocyst, with a marked decrease at the 4-cell stage. To determine whether the temporal pattern of enzyme activity was dependent on the maternal to zygotic transition, embryos were incubated in the presence of a transcription inhibitor, α-amanitin. The reappearance of the enzyme activity after the 4-cell stage was blocked in α-amanitin treated embryos, indicating the requirement of embryonic transcription. On the other hand, the embryonic steroid metabolism was tested by incubating blastocyst with tritiated pregnenolone. Analysis of the metabolites by TLC indicated the production of a compound with a mobility identical to progesterone. These results described the expression of the 3β-HSD and the activity of this metabolic enzyme in bovine oocytes and preimplantation embryos, suggesting that steroids may act as autocrine effectors on preimplantation embryo development. © 2002 Wiley-Liss, Inc. |
| format |
JOUR |
| author |
Chiappe, M.E. Lattanzi, M.L. Colman-Lerner, A.A. Barañao, J.L. Saragüeta, P. |
| author_facet |
Chiappe, M.E. Lattanzi, M.L. Colman-Lerner, A.A. Barañao, J.L. Saragüeta, P. |
| author_sort |
Chiappe, M.E. |
| title |
Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| title_short |
Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| title_full |
Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| title_fullStr |
Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| title_full_unstemmed |
Expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| title_sort |
expression of 3β-hydroxysteroid dehydrogenase in early bovine embryo development |
| url |
http://hdl.handle.net/20.500.12110/paper_1040452X_v61_n2_p135_Chiappe |
| work_keys_str_mv |
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| _version_ |
1807322115191341056 |