Nuclear transfer technique affects mRNA abundance, developmental competence and cell fate of the reconstituted sheep oocytes

• DOI: 10.1530/rep-12-0318
• Publication Date: 2013-02-12T03:25:25Z
• Authors: F Moulavi; S M Hosseini; M Hajian; M Forouzanfar; P Abedi; S Ostadhosseini; V Asgari; M H Nasr-Esfahani
• Keywords: 2. Zero hunger; Nuclear Transfer Techniques; 0303 health sciences; 03 medical and health sciences; Sheep; Oocytes; Animals; Embryonic Development; Female; RNA, Messenger; Real-Time Polymerase Chain Reaction

The effect of technical steps somatic cell nuclear transfer (SCNT) on different aspects cloned embryo development was investigated in sheep. In vitro-matured oocytes were enucleated the presence or absence zona and reconstituted by three SCNT techniques: conventional zona-intact (ZI-NT), standard zona-free (ZF-NT) intracytoplasmic injection (ICI-NT). Stepwise alterations remodeling events mRNA abundances, throughput efficiency allocation resulted blastocysts assessed. Early signs observed as soon 2 h post-reconstitution (hpr) for fusion-based methods (ZI-NT ZF-NT) but not observable until 4 hpr with ICI-NT method. relative abundances HSP90AA1 (HSP90), NPM2 ATPase genes affected i) zona, ii) oocyte enucleation method iii) After reconstitution, however, contents POU5F1 (OCT4) ZI-NT ZF-NT PAPOLA (PAP) significantly lower than those Zona removal doubled blastocyst technique compared ICI-NT. Cleavage rate protocol, whereas yield (17.0±1.0%) (P<0.05; ANOVA) higher (7.1±1.5%) group (11.2±3.3%). Despite similarities total number, protocol changed distribution cells blastocysts, ZF-NT-cloned had smaller inner mass ZI-NT. These results indicate that cloning may result variety phenotypes.