Proteins Involved in the LH Switch Are Present in Detergent-Resistant Membranes of Porcine Granulosa Cells but Not in Cumulus-Oocyte Complexes.
Notice bibliographique
Résumé
Lipids rafts are specialized membrane microdomains constituted of sphingolipids, cholesterol and proteins forming platforms that function in membrane signalling and trafficking. Membrane signalling and trafficking are important processes taking place in the ovarian follicle leading to different physiological events such as ovulation. We hypothesized that molecules involved in the Luteinizing Hormone (LH) switch leading to ovulation such as Epidermal Growth Factor (EGF) receptor, progesterone receptor, prostaglandin E2 receptor, Connexin 43 and LH receptor are present in lipid rafts of granulosa cells and cumulus-oocyte complexes (COCs). It is known that lipid rafts can be isolated as detergent-resistant membranes (DRMs) in a cell homogenate. In the present study, two methodological strategies have been used to isolate DRMs present in porcine granulosa cells and COCs from 2-6 mm diameter follicles. In method 1, granulosa cells and COC samples were exposed to Triton X-100 at 4°C and DRMs were isolated by ultracentrifugation of cell extracts in a sucrose density gradient. In method 2, DRMs were isolated by a differential detergent extraction approach. Then, the molecules of interest were detected in the DRMs isolated by method 1 and 2 by Dot Blot assay using specific antibodies. Both methodological strategies allowed the isolation of DRMs as shown by the presence of the ganglioside GM1, a lipid raft marker, in the DRMs isolated by method 1 and 2. However, the results will be discussed based on method 2 (differential detergent extraction) since the contamination of DRMs by membranes from the nucleus and Golgi apparatus is largely decreased in this method. To support the isolation of DRMs, the protein Caveolin-1, a lipid raft marker, was also present in DRMs isolated from both granulosa cells and COCs while the protein Flotillin-1, another lipid raft marker, was only present in granulosa cells. The transferrin receptor, which is known to be cytosolic, was absent in the DRMs isolated from both granulosa cells and COCs, supporting the correct isolation of DRMs.All candidates involved in the LH switch (EGF receptor, progesterone receptor, prostaglandin E2 receptor, Connexin 43 and LH receptor) were present in the DRMs isolated from granulosa cells while only Connexin 43 was present in the DRMs isolated from COCs. In conclusion, DRMs are present in both porcine granulosa cells and COCs. Interestingly, proteins involved in the LH switch are present in lipid rafts of granulosa cells but not in those of COCs suggesting a specific role of lipid rafts as a platform for cell signalling in the ovarian follicle. (poster)
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 | 0,001 | 0,000 |
| 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 ».