Cargando…
Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes
BACKGROUND: The competence for embryonic development after IVF is low in the yak, therefore, we investigated the effects of supplementation of FSH, LH and the proteasome inhibitor MG132 in IVM media on yak oocyte competence for development after IVF. METHODS: In Experiment 1, yak cumulus-oocyte comp...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3998235/ https://www.ncbi.nlm.nih.gov/pubmed/24754924 http://dx.doi.org/10.1186/1477-7827-12-30 |
_version_ | 1782313317789859840 |
---|---|
author | Xiao, Xiao Zi, Xiang-Dong Niu, Hui-Ran Xiong, Xian-Rong Zhong, Jin-Cheng Li, Jian Wang, Li Wang, Yong |
author_facet | Xiao, Xiao Zi, Xiang-Dong Niu, Hui-Ran Xiong, Xian-Rong Zhong, Jin-Cheng Li, Jian Wang, Li Wang, Yong |
author_sort | Xiao, Xiao |
collection | PubMed |
description | BACKGROUND: The competence for embryonic development after IVF is low in the yak, therefore, we investigated the effects of supplementation of FSH, LH and the proteasome inhibitor MG132 in IVM media on yak oocyte competence for development after IVF. METHODS: In Experiment 1, yak cumulus-oocyte complexes (COCs) were in vitro matured (IVM) in TCM-199 with 20% fetal calf serum (FCS), 1 microg/mL estradiol-17beta, and different combinations of LH (50 or 100 IU/mL) and FSH (0, 1, 5, 10 microg/mL) at 38.6 degrees C, 5% CO(2) in air for 24 h. Matured oocytes were exposed to frozen–thawed, heparin-capacitated yak sperm. Presumptive zygotes were cultured in SOF medium containing 6 mg/ml BSA, 0.5 mg/mL myoinositol, 3% (v/v) essential amino acids, 1% nonessential amino acids and 100 μg/mL L-glutamine (48 h, 38.5 degrees C, 5% CO(2), 5% O(2), and 90% N(2)). In Experiment 2, cumulus cells were collected at the end of IVM to determine FSHR and LHR mRNA expression by real-time PCR. In Experiment 3 and 4, COCs were cultured in the presence or absence of the proteasomal inhibitor MG132 from either 0–6 h or 18–24 h after initiation of maturation. RESULTS: The optimum concentration of FSH and LH in IVM media was 5 microg/mL FSH and 50 IU/mL LH which resulted in the greatest cleavage (79.1%) and blastocyst rates (16.1%). Both FSHR and LHR mRNA were detected in yak cumulus cells after IVM. Treatment with MG132 early in maturation reduced (P < 0.05) cleavage and blastocyst rates. Conversely, treatment with MG132 late in maturation improved (P < 0.05) blastocyst rate. Optimal results with MG132 were achieved at a concentration of 10 microM. CONCLUSIONS: An optimum concentration of FSH and LH in IVM medium, and treatment with MG132 late in maturation can improve yak oocytes competence for development after IVF. |
format | Online Article Text |
id | pubmed-3998235 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-39982352014-04-25 Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes Xiao, Xiao Zi, Xiang-Dong Niu, Hui-Ran Xiong, Xian-Rong Zhong, Jin-Cheng Li, Jian Wang, Li Wang, Yong Reprod Biol Endocrinol Research BACKGROUND: The competence for embryonic development after IVF is low in the yak, therefore, we investigated the effects of supplementation of FSH, LH and the proteasome inhibitor MG132 in IVM media on yak oocyte competence for development after IVF. METHODS: In Experiment 1, yak cumulus-oocyte complexes (COCs) were in vitro matured (IVM) in TCM-199 with 20% fetal calf serum (FCS), 1 microg/mL estradiol-17beta, and different combinations of LH (50 or 100 IU/mL) and FSH (0, 1, 5, 10 microg/mL) at 38.6 degrees C, 5% CO(2) in air for 24 h. Matured oocytes were exposed to frozen–thawed, heparin-capacitated yak sperm. Presumptive zygotes were cultured in SOF medium containing 6 mg/ml BSA, 0.5 mg/mL myoinositol, 3% (v/v) essential amino acids, 1% nonessential amino acids and 100 μg/mL L-glutamine (48 h, 38.5 degrees C, 5% CO(2), 5% O(2), and 90% N(2)). In Experiment 2, cumulus cells were collected at the end of IVM to determine FSHR and LHR mRNA expression by real-time PCR. In Experiment 3 and 4, COCs were cultured in the presence or absence of the proteasomal inhibitor MG132 from either 0–6 h or 18–24 h after initiation of maturation. RESULTS: The optimum concentration of FSH and LH in IVM media was 5 microg/mL FSH and 50 IU/mL LH which resulted in the greatest cleavage (79.1%) and blastocyst rates (16.1%). Both FSHR and LHR mRNA were detected in yak cumulus cells after IVM. Treatment with MG132 early in maturation reduced (P < 0.05) cleavage and blastocyst rates. Conversely, treatment with MG132 late in maturation improved (P < 0.05) blastocyst rate. Optimal results with MG132 were achieved at a concentration of 10 microM. CONCLUSIONS: An optimum concentration of FSH and LH in IVM medium, and treatment with MG132 late in maturation can improve yak oocytes competence for development after IVF. BioMed Central 2014-04-22 /pmc/articles/PMC3998235/ /pubmed/24754924 http://dx.doi.org/10.1186/1477-7827-12-30 Text en Copyright © 2014 Xiao et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Xiao, Xiao Zi, Xiang-Dong Niu, Hui-Ran Xiong, Xian-Rong Zhong, Jin-Cheng Li, Jian Wang, Li Wang, Yong Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title | Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title_full | Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title_fullStr | Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title_full_unstemmed | Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title_short | Effect of addition of FSH, LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes |
title_sort | effect of addition of fsh, lh and proteasome inhibitor mg132 to in vitro maturation medium on the developmental competence of yak (bos grunniens) oocytes |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3998235/ https://www.ncbi.nlm.nih.gov/pubmed/24754924 http://dx.doi.org/10.1186/1477-7827-12-30 |
work_keys_str_mv | AT xiaoxiao effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT zixiangdong effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT niuhuiran effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT xiongxianrong effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT zhongjincheng effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT lijian effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT wangli effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes AT wangyong effectofadditionoffshlhandproteasomeinhibitormg132toinvitromaturationmediumonthedevelopmentalcompetenceofyakbosgrunniensoocytes |