Bibliographic record
Abstract
Introduction:We have described the significance of sperm surface P68 in sperm-zona pellucida (ZP) binding.P68 peptide sequencing and molecular cloning identify P68 as arylsulphatase A (AS-A), generally recognized as a lysosomal enzyme.Our objective was to demonstrate the presence of AS-A on the sperm head surface and its participation in gamete interaction. Materials and methods: IgG fraction was purified from rabbit anti-AS-A antiserum, generated against human liver AS-A, by protein A chromatography.Immunoelectronmicroscopy was performed in live mouse and pig sperm, treated with anti-AS-A IgG.The function of sperm surface AS-A was assessed by in-vitro gamete binding assays using anti-AS-A IgG-treated sperm.In addition, the in-vivo fertilization rate in mice inseminated with anti-AS-A IgG-treated sperm was determined.Sperm treated with preimmune rabbit IgG served as controls in all experiments.Direct binding of AS-A, conjugated with Alexa 430 fluorochrome, to intact and solubilized ZP was also determined and K d of the binding was analysed by Scatchard plotting.A competitive binding assay was performed to determine the specificity of pig ZP sulphoglycoprotein to which AS-A bound.All experiments were performed more than three times and results from treated samples were analysed for significant differences from the control values using Student's t-test.Results: Immunoelectronmicroscopy revealed gold labelling of AS-A over the sperm head surface.Pretreatment of sperm with anti-AS-A antibody resulted in dose-dependent inhibition of sperm-ZP binding, with the maximum inhibition (~75%) at 100 µg/ml anti-AS-A IgG.Furthermore, a marked inhibition (~60% average from five experiments) of IVF in female mice inseminated with 200 µg/ml anti-AS-A IgG was observed.We also demonstrated that Alexa-430 AS-A bound directly to both intact and solubilized pig and mouse ZP, with K d of 0.50 and 0.21 µmol/l respectively.These values were in same range as that observed for mouse sperm-ZP binding, supporting the concept that AS-A participated in sperm-ZP interaction.Furthermore, competitive binding assays revealed that pig ZP3α sulphoglycoprotein, a major player in sperm binding, inhibited AS-A-ZP binding to 15% of the control value at only 2 µmol/l.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.002 | 0.001 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.004 | 0.002 |
| Insufficient payload (model declined to judge) | 0.674 | 0.448 |
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 itClassification
machine, unvalidatedMachine predicted; the direct Gemma label and the distilled Codex classifier agree on what is shown here.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".