Background
Methods
Oocyte collection, in vitro maturation, in vitro fertilization and embryo culture
Temporal changes in BMP system mRNA abundance in bovine oocytes and early embryos
Gene | Genbank accession number | Primer sequence | Size (bp) |
---|---|---|---|
BMP2
| BC134682 | F: 5′-AAGGCCCTTGCTTGTCACTTT-3′ | 72 |
R: 5′-TGCTTGCCGCTTTTCTCTTC-3′ | |||
BMP3
| XM_587912 | F: 5′-ATCTGTGGCTGAGCTGCTTGT-3′ | 62 |
R: 5′-GGAAGGGCTGCCTGAGTCT-3′ | |||
BMP7
| XM_612246 | F: 5′-TGCCACTAGCTCTTCCTGGAA-3′ | 65 |
R: 5′-TGAGAGACCCAGGATCCAGAA-3′ | |||
BMP10
| XM_583418 | F: 5′-CGCCCACGAGCAATTCC-3′ | 66 |
R: 5′-TCCCCAGGTCCGTTGGA-3′ | |||
SMAD1
| BC116117 | F: 5′-CACCATGAACTGAAACCATTGG-3′ | 68 |
R: 5′-GATGCACACCTCCTTCTGCTT-3′ | |||
SMAD5
| DV821574 | F: 5′-GCAACGTTTCCTGATTCTTTCC-3′ | 74 |
R: 5′-GGCGGGTAGGGACTATTTGG-3′ | |||
ALK3
| NM_001076800 | F: 5′-TCAGCGAACTATTGCCAAACAG-3′ | 75 |
R: 5′-CCCATCCACACTTCTCCGTATC-3′ | |||
ALK6
| NM_001105328 | F: 5′-CCCACCCCTCGTCCAAAG-3′ | 63 |
R: 5′-GACCGAGTCTTCTGGACAATGG-3′ | |||
ALK2
| BC133311 | F: 5′-TTGGCCTCATCATTTTGTCTGT-3′ | 70 |
R: 5′-CGGAGAGCAACTCCCAATAGG-3′ | |||
ACVR2A
| NM_174227 | F: 5′- CCACAAACCCGCCATATCTC -3′ | 185 |
R: 5′- TAGCACCCTCTAACACCTCTG -3′ | |||
ACVR2B
| NM_174495 | F: 5′-GGAGCCATCAACTTCCAGAG-3′ | 121 |
R: 5′-GCATGTACTCATCCACAGGTC-3′ | |||
BMPR2
| XM_002685492 | F: 5′-AACACCACTCAGTCCACCTC-3′ | 120 |
R: 5′-GTCAGCATCCTATATCCAAAGCA-3′ | |||
NANOG
| NM_001025344 | F: 5′-AAAGTTACGTGTCCTTGCAAACG-3′ | 73 |
R: 5′-GAGGAGGGAAGAGGAGAGACAGT-3′ | |||
CDX2
| AM293662 | F: 5′-FCGTCTGGAGCTGGAGAAGGA-3′ | 70 |
R: 5′-CGGCCAGTTCGGCTTTC-3′ |
Effect of transcriptional inhibition on BMP system mRNA abundance
Effect of BMP2 supplementation on bovine embryo developmental progression
Statistical analysis
Results and discussion
Temporal regulation of BMP mRNA abundance during oocyte maturation and early embryogenesis
Effect of embryo culture in the presence of the transcriptional inhibitor α-amanitin on BMP, BMP receptor and SMAD1 and SMAD5 mRNA abundance
Gene | RT with oligo dT | RT with random hexamers | ||
---|---|---|---|---|
Control | α-Amanitin treated | Control | α-Amanitin treated | |
SMAD1
| 1.15 ± 0.27a | 9.08 ± 1.19b | 1.03 ± 0.16 | 1.77 ± 0.17 |
SMAD5
| 1.17 ± 0.35a | 7.33 ± 1.27b | 1.02 ± 0.12 | 1.45 ± 0.17 |
BMP2
| 1.10 ± 0.27a | 3.19 ± 0.76b | 1.25 ± 0.49 | 1.30 ± 0.21 |
BMP3
| 1.02 ± 0.11a | 2.85 ± 0.60b | 1.15 ± 0.36 | 3.26 ± 0.88 |
BMP7
| 1.02 ± 0.12a | 3.36 ± 0.66b | 1.17 ± 0.40 | 2.96 ± 1.31 |
BMP10
| 1.04 ± 0.19a | 3.82 ± 0.25b | 1.07 ± 0.23 | 1.59 ± 0.31 |
ALK3
| 1.23 ± 0.37a | 9.41 ± 0.72b | 1.06 ± 0.22 | 1.42 ± 0.31 |
ALK6
| 1.12 ± 0.25a | 5.43 ± 0.71b | 1.05 ± 0.20 | 1.38 ± 0.16 |
ALK2
| 1.27 ± 0.50a | 5.73 ± 1.34b | 1.02 ± 0.13 | 0.86 ± 0.09 |