97 Function of species-specific OCT4 reporter system during porcine embryo development
Notice bibliographique
Résumé
The present study examined the activity and function of pig OCT4 (POU5F1) enhancer in porcine early embryo development. OCT4 is one of the master regulators for the pluripotency of early mammalian embryonic development and embryonic stem cells. It has two regulatory elements: distal enhancer (DE) and proximal enhancer (PE) in the upstream regulatory region. They are activated under different conditions such as naïve and primed. It is known that two enhancers are activated to produce OCT4 simultaneously or sequentially depending on the state of pluripotency. Many porcine OCT4 upstream region-based reporter systems have been reported because this is a necessary part of studying porcine-specific pluripotency. However, the porcine-specific OCT4 reporter system has never been transfected and functionally tested during porcine early embryo development. We performed functional tests of the previously established porcine-specific OCT4 reporter system in the early embryo development stage. Porcine embryos were micro-injected with the pOCT4-ΔPE-eGFP (DE-GFP) containing a distal enhancer and core promoter and pOCT4-ΔDE-DsRed2 (PE-RFP) containing a proximal enhancer and core promoter. They were cultured in PZM-3 at 39°C in a humidified atmosphere, 5% CO2, and 5% O2 for 168 h. Fifty of the 100 embryos were injected with OCT4 reporter system as treatment groups and the other 50 were not injected (control groups). We analysed mRNA and protein expression of GFP and RFP using quantitative real-time PCR and confocal microscopy by developmental stages. The introduced reporter system could function beginning with the 4-cell stage. The expression of GFP and RFP was observed simultaneously in the early embryonic development stage up to blastocyst, indicating that porcine OCT4 was produced by distal and proximal enhancers, unlike mouse Oct4 expression, which was only controlled by the distal enhancer during early embryo development. Therefore, the mechanisms and functions of distal and proximal enhancers of porcine OCT4 were different from those of the mouse. This is similar to results of the previous experiment using porcine-induced pluripotent stem cells, which suggest that porcine OCT4 is expressed by two enhancers and in a stage-specific manner during reprogramming and that pigs do not use only one enhancer in pluripotent states, but two enhancers at the same time, and there is only a difference in degree. This work showed that the porcine OCT4 reporter system enables non-destructive analysis during early embryo development and it could be applied to study species-specific pluripotency and to help the establishment of naïve embryonic stem cells in the pig. This work was supported by the BK21 Plus Program, the National Research Foundation of Korea (NRF) grant funded by the Korea government (NRF-2019R1C1C1004514), the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-3-HD020).
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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 ».