Assessment by differential display‐RT‐PCR of mRNA transcript transitions and α‐amanitin sensitivity during bovine preattachment development
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Bibliographic record
Abstract
The objectives of this study were to compare patterns of mRNA expression, investigate the onset of transcription, and isolate stage-specific and α-amanitin-sensitive mRNAs during early bovine development by differential-display-reverse transcription-polymerase chain reaction (DD-RT-PCR). Embryos representing a preattachment developmental series from the 1-cell to the expanded/hatched blastocyst stage were cultured in synthetic oviduct fluid medium + citrate and amino acids (cSOFMaa) with and without α-amanitin (100 μg/mL) for 4 and 12 hr. mRNA profiles were displayed by DD-RT-PCR using 5′ primers A and N. Total conserved cDNA banding patterns varied according to embryo stage with cDNA band numbers declining during early cleavage stages compared to oocyte values and then increasing in total number from the 6–8-cell stage through to the blastocyst stage. A cDNA banding pattern was established at the 8–16-cell stage that was largely unchanged through to the blastocyst stage. These findings with respect to cDNA banding patterns were conserved between oligo primer sets and experimental replicates. α-Amanitin sensitivity was first detected at the 2–5-cell stage but became predominant following the 6–8-cell stage of development to eventually affect the appearance of up to 40% of all cDNA bands by the blastocyst stage. A 12 hr α-amanitin treatment was required to effectively block 3H-uridine incorporation into mRNA in blastocyst stage embryos. Several stage-specific and α-amanitin-sensitive cDNAs were isolated and they will be a focus for future studies. In conclusion, DD-RT-PCR is an effective tool for contrasting gene expression patterns and isolating uncharacterized mRNA transcripts during bovine early development. Mol. Reprod. Dev. 55:152–163, 2000. © 2000 Wiley-Liss, Inc.
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Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it