1 PATHWAYS AND CELLULAR FUNCTIONS INFLUENCED BY INSULIN TREATMENT DURING OOCYTE MATURATION – A TRANSCRIPTOME STUDY OF IN VITRO-PRODUCED BOVINE DAY 8 BLASTOCYSTS
Notice bibliographique
Résumé
Insulin as a key metabolic hormone has crucial functions in metabolic regulation in all mammals. Deviation of its physiological concentration occurs in metabolic disorders as obesity and diabetes in humans or negative energy balance and overfeeding in the cow. As these metabolic disorders are strongly correlated with reproductive disturbances, we investigated the effect of insulin during oocyte maturation on gene expression of bovine Day 8 blastocysts (BC8) by transcriptome analysis. Abattoir-derived oocytes (n = 882) were divided into 3 groups and in vitro matured for 22 h by adding insulin (H: High 10 µg mL–1; L: Low 0.1 µg mL–1 and Z: Zero, control). This was followed by standard in vitro production (IVP) and evaluation of developmental rates up to blastocyst stage. BC8 (n = 120) were pooled in groups of 10 and total RNA was extracted by parallel gDNA and total RNA-extraction (AllPrepDNA/RNA micro kit, cat no. 80284, Qiagen®, Valencia, CA, USA) for analyses of the transcriptome. All samples (4 biological replicates/group) resulted in RIN-values >7.5. RNA amplification, cDNA synthesis, purification, and labelling were performed and 825 ng of Cy3- and Cy5-labelled linearly amplified aRNA was hybridized on the Agilent-manufactured EmbryoGENE-slides in a 2-colour dye swap design. An empirical Bayes moderated t-test was applied to search for the differentially expressed transcripts (DET) between control and insulin-treated groups, using the ‘limma’ package in R (www.r-project.org). The DET were defined as having a 1.5-fold change difference between treatment and control and P < 0.05. Pathways and molecular functions influenced by insulin treatment were analysed by using Ingenuity Pathway Analysis (IPA; Ingenuity® Systems, www.ingenuity.com). As a global pattern, insulin treatment induced an up-regulation of genes. In total, 202 DET in the H and 142 DET in the L group were found where 104 DET were common in both insulin groups. Fifteen selected candidate genes chosen for qPCR validation and 12 (80%) showed similar expression patterns as the microarray data. DET relevant for following cellular functions were found in H: Cell Cycle, Cellular Compromise, Lipid Metabolism, Molecular Transport, Small Molecule Biochemistry respective L: Cell Morphology, Cellular Growth and Proliferation, Cell Cycle, Carbohydrate Metabolism and Cellular Assembly and Organization. The top canonical pathways influenced were Epithelial Adherens Junction Signalling and Remodelling, Germ Cell Sertoli Cell Junction Signalling and NRF2-mediated Oxidative Stress Response. Correlatively, blastocyst rates on Day 8 were significantly lower in H and L v. Z (P < 0.05). The transcriptome data could explain the mechanisms behind the impaired development, as genes involved in cellular growth and energy metabolism in Day 8 blastocysts were affected. The fact that transcripts related to NRF2-mediated oxidative stress response and lipid metabolism are up-regulated suggests that insulin induces dysregulation of cellular functions and energy metabolism leading to impaired embryo developmental potential. Funded by FORMAS.
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 machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 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) | 0,001 | 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 source (Gemma direct ou Codex distillé), 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 ».