Evaluation of In Vitro Capacitation of Buffalo Frozen/Thawed Sperm by Different Techniques
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Bibliographic record
Abstract
This study aimed to determine the most reliable method to evaluate capacitation of buffalo frozen/thawed sperm. Frozen/thawed sperm cells were incubated in Tyrode albumin lactate pyruvate medium (TALP) in absence of capacitating agents (control) and in presence of 10 µg/ml heparin for 2 and 4 h. Capacitation was assessed by Trypan blue/Giemsa after lysophosphatidylcholine (LPC) exposure, chlortetracycline (CTC) fluorescence assay and immune-localization of tyrosine phosphorylated protein. Furthermore, we evaluated the effect of heparin on penetration, cleavage rates and kinetics of embryo development after heterologous IVF. The percentage of LPC-induced acrosome reacted (AR)-sperm increased (P<0.05) with heparin compared to the control after 2 h (28.2 vs 24.4%, respectively) and 4 h (35.1 vs 32.0 %, respectively). No differences in CTC pattern B (capacitated sperm) were found between groups and incubation times (on average 63%). On the contrary, heparin decreased (P<0.01) the percentage of tyrosine phosphorylation pattern A after 2 and 4 h (34.3 and 35.3%, respectively) compared to the control (54.5 and 51.8%, respectively) and increased (P<0.01) that of pattern EA after 2 and 4 h (59.2 and 54.2 %, respectively) compared to the control group (44.7 and 45.2 %, respectively). Both cleavage and penetration rates, as well as the percentage of fast developing embryos, were higher (P<0.01) in the heparin-treated group (77.2, 80.4 and 74.0 %, respectively) compared to the control (56.6, 58.0 and 55.2 %, respectively). In conclusion, Trypan blue/Giemsa staining to evaluate LPC-induced AR and tyrosine protein phosphorylation assay can be successfully used to evaluate capacitation of buffalo frozen/thawed semen.
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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 | 0.004 | 0.002 |
| 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