Tissue Specific Transcription Start Site and Alternative Splicing of the MAESTRO Gene (MRO): in Granulosa Cells from PCOS and Non-PCOS Patients, and in Control Non-Ovarian Tissues.
Bibliographic record
Abstract
MRO, whose gene product and function remains unknown, was first described during male murine gonadal somatic cell differentiation, but not in the developing female gonads. We previously showed that MRO has a remarkable over expression in granulosa cells from lean patients with Polycystic Ovarian Syndrome (PCOS). The aim of the study was to 1) investigate the different splice variants expression patterns of the MRO gene in human ovarian granulosa cells from PCOS and non-PCOS patients. 2) To characterize tissue specific transcription start sites of the human MRO gene. 3) To isolate the MRO protein from human tissues. The project was approved by the Research Ethics board of Sunnybrook Health Sciences Centre and all participants provided informed consent. Total RNA was isolated from granulosa lutein cells obtained from patients undergoing in vitro fertilization (IVF) and from other commercially acquired human tissues. Gene expression was analyzed by RT-PCR. Whole protein extracts from granulosa cells were resolved by SDS-PAGE and analyzed by Western blot using a polyclonal antibody raised against the deduced peptide. The presence of multiple splice variants in several human tissues was demonstrated using RT-PCR. A unique MRO transcript containing the non-coding exon 1 was found exclusively in GLC and testis, but not in other tissues. The two main variants were cloned into an expression vector in order to express the protein in a cell-free system. Western blot analysis on whole protein extracts from granulosa cells revealed two bands -- 29kDa and 26kDa). The 26kDa appeared also with the expression vector analysis. In conclusion, we show evidence that a unique transcript of the human MRO gene, containing the non-coding region of exon 1, but not exon 2, is expressed in human adult granulosa lutein cells and testis but not in other tissues. The transcript containing exon 1 encodes an alternative promoter which may serve as a tissue-specific regulatory sequence. We are currently attempting to express and purify the different protein isoforms. This contributes to our previous findings of differential MRO expression in human cumulus cells from lean PCOS patients undergoing IVF compared to their non-PCOS counterparts. Further studies of MRO may reveal novel processes involved in both normal ovarian physiology and in PCOS. (poster)
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How this classification was reachedexpand
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 | 0.001 | 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 itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
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".