Transcriptome Analysis of Pig In Vivo, In Vitro-Fertilized, and Nuclear Transfer Blastocyst-Stage Embryos Treated with Histone Deacetylase Inhibitors Postfusion and Activation Reveals Changes in the Lysosomal Pathway.
Cell Reprogram
; 17(4): 243-58, 2015 Aug.
Article
in En
| MEDLINE
| ID: mdl-26731590
ABSTRACT
Genetically modified pigs are commonly created via somatic cell nuclear transfer (SCNT). Treatment of reconstructed embryos with histone deacetylase inhibitors (HDACi) immediately after activation improves cloning efficiency. The objective of this experiment was to evaluate the transcriptome of SCNT embryos treated with suberoylanilide hydroxamic acid (SAHA), 4-iodo-SAHA (ISAHA), or Scriptaid as compared to untreated SCNT, in vitro-fertilized (IVF), and in vivo (IVV) blastocyst-stage embryos. SAHA (10 µM) had the highest level of blastocyst development at 43.9%, and all treatments except 10 µM ISAHA had the same percentage of blastocyst development as Scriptaid (p<0.05). Two treatments, 1.0 µM ISAHA and 1.0 µM SAHA, had higher mean cell number than No HDACi treatment (p<0.021). Embryo transfers performed with 10 µM SAHA- and 1 µM ISAHA-treated embryos resulted in the birth of healthy piglets. GenBank accession numbers from up- and downregulated transcripts were loaded into the Database for Annotation, Visualization and Integrated Discovery to identify enriched biological themes. HDACi treatment yielded the highest enrichment for transcripts within the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway, lysosome. The mean intensity of LysoTracker was lower in IVV embryos compared to IVF and SCNT embryos (p<0.0001). SAHA and ISAHA can successfully be used to create healthy piglets from SCNT.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Blastocyst
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Fertilization in Vitro
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Sus scrofa
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Nuclear Transfer Techniques
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Histone Deacetylase Inhibitors
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Transcriptome
Limits:
Animals
Language:
En
Journal:
Cell Reprogram
Year:
2015
Document type:
Article
Publication country:
EEUU
/
ESTADOS UNIDOS
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ESTADOS UNIDOS DA AMERICA
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EUA
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UNITED STATES
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UNITED STATES OF AMERICA
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US
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USA