3 X-LINKED GENE EXPRESSION IN BOVINE PRE-IMPLANTATION EMBRYOS OBTAINED IN VIVO AND IN VITRO AS A MEASURE OF IMPACT OF EMBRYO PRODUCTION TECHNOLOGIES
Notice bibliographique
Résumé
The X chromosome provides an ideal system to study the impact of assisted reproduction technologies on gene expression because it holds a wide range of diversely functional genes and also the epigenetic features of X-inactivation are inherently susceptible to in vitro culture-mediated alterations. Using quantitative real-time RT-PCR we studied the expression of a panel of X-linked genes (ANT3, GAB3, RPS4, MECP2, XIAP, XIST) in pooled pre-attachment bovine embryos produced in vivo and in vitro. We collected pools of ten embryos in vivo (de la Fuente R. et al. 1999 Biol. Reprod. 60, 769–775) at the morula (n = 3 pools) and blastocyst (n = 3 pools) stages. Pools of ten matured oocytes (n = 10 pools), 2–4 cell (n = 10 pools), 8–16 cell (n = 10 pools), morula (n = 7 pools) and blastocyst (n = 7 pools) stage embryos were produced in LSOF (Robert et al. Biol. Reprod. 67, 1465–1472) and TCM-199/bovine oviductal epithelial cells (BOEC) co-culture (de la Fuente et al. 1999 Biol. Reprod. 60, 769–775). Total RNA was extracted with the Absolutely RNA® Microprep kit (Stratagene, La Jolla, CA, USA) and reverse transcribed using Oligo-dT12–18 primers and Superscript II RT (Invitrogen, Burlington, Ontario, Canada). Specific primers were designed for each gene and PCR products were used to build standard curves for absolute quantification in a Light Cycler™ instrument with the Light Cycler FastStart DNA Master Mix SYBRGreenI kit (Roche Diagnostics, Laval, Quebec, Canada). The data were analyzed with SAS (SAS Institute Inc., Cary, NC, USA) using a factorial ANOVA design and a log transformation. Significance level was set at a = 0.05. There were no significant differences in transcript levels between IVF systems in mature oocytes, morulae, or blastocysts. Significant differences were observed for some of the genes tested at the 2–4-cell (XIAP) and at the 8–16-cell stages (ANT3, GAB3, XIAP) but not others. There were significant differences between IVF (both BOEC and LSOF) and in vivo embryos at the morula and blastocyst stages, with IVF embryos showing an average of 8 (ANT3), 10 (XIAP), 127 (MECP2), and 593 (XIST) times more transcripts than their in vivo counterparts. GAB3 was detected in only a few samples prior to the blastocyst stage where it was consistently detected in IVF but not in vivo embryos. It was concluded that there is an effect of the IVF system on gene transcription just before and during the period of activation of the embryonic genome during the 4th cell cycle, as well as marked differences between IVF and in vivo produced embryos that are evident in the differential expression of X-linked genes. This system provides a good tool to monitor the effects of embryo production conditions and help in their improvement. This project was supported with grants from NSERC, FSBC, and OMAF.
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 ».