Effect of Butirolactone I on the gene expression of PTGS2, GREM1 and PFKP of the cumulus cells in bovine in vitro matured oocytes
Notice bibliographique
Résumé
A small proportion of bovine in vitro matured oocytes (IVM) has the potential to develop to embryo. Aiming to improve oocyte competence, some inhibitors of cyclin-dependent kinases, such as butirolactone I (BL), have been used to synchronize the nuclear and cytoplasmic maturation of bovine oocytes. Moreover, the cumulus cells (CCs) also play an important role for the acquisition of in vitro oocyte competence. Therefore, the aim of this study was to investigate the gene expression of PTGS2, GREM1 and PFKP in the CCs of in vitro or in vivo matured (IVOM) bovine oocytes. Ovaries were collected from a local slaughterhouse, and the oocytes from the follicles were obtained. Oocytes were randomly divided into 5 groups: I (immature oocytes); II (IVOM); III (IVM); IV (meiosis block, 12h; reversal, 12h); and V (meiosis block, 24h; reversal, 24h). All reagents were purchased from SigmaAldrich (St. Louis, USA), unless otherwise specified. The cumulus-oocytes complexes (COCs) were washed and selected in PBS medium plus 10% FCS (Nutricell®, Campinas, Brazil). The medium 199, supplemented with sodium bicarbonate, sodium pyruvate, penicillin, FSH and LH (Bioniche Inc., Canada), estradiol, cisteamine and 10% FCS was used to IVM. The IVM medium, supplemented with 10 µM of BL, without hormones and FCS, was used to meiosis block. For the IVOM, the estrus of 10 Nelore donor cows were synchronized with progesterone (Primer®, Tecnopec, Brazil), estradiol benzoate (Sincrocio®, Ourofino, Brazil) and cloprostenol sodium (Ciosin®, MSD, Brazil). For super stimulation of the ovaries, FSH was injected (six decreasing doses), and final follicle maturation was induced with gonadorelin (Fertagyl®, Intervet, Brazil). The COCs were obtained through OPU, 19- 20 h after gonadorelin injection. The CCs were removed by several pipetting in PBS medium with 0.1% hyaluronidase. Then, droplets containing only CCs were centrifuged (3,355 x g/10 min) and frozen in total RNA extraction medium. Gene expression was investigated by quantitative RT-PCR, normalized by GAPDH constitutive gene. The results were analyzed using ANOVA and Tukey’s test, with P<0.05 taken as significant. There was no statistical difference among the groups for PTGS2. However, the abundance of transcripts for GREM1 was higher (P<0.05) in the CCs from oocytes from group II than from the other groups. The CCs from group II showed higher (P<0.05) abundance of transcripts for PFKP compared to groups I and IV, with similar results for groups I and IV. In conclusion, the meiosis block with BL did not affect the abundance of transcripts of any gene investigated, however, as a higher GREM1 expression was observed in the CCs from IVOM oocytes, further researches using different compounds for IVM medium may identify pivotal factors that are related with the expression of these genes in CCs.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,002 | 0,002 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».