Relative expression of cell growth regulatory genes insulin-like growth factors (IGF-1 and IGF-2) and their receptors (IGF-1R and IGF-2R) in somatic cell nuclear transferred (SCNT) and in vitro fertilized (IVF) pre-implantation buffalo embryos.
Relative mRNA transcript expression of insulin-like growth factors, IGF-1, IGF-2 and their receptors, IGF-1R and IGF-2R, was studied in SCNT and IVF buffalo embryos at different developmental stages using SYBR green with real-time PCR. SCNT embryos were produced by enucleating IVM oocytes and transferring granulosa cells (passage 5) followed by the electrofusion and chemical activation method. IVF embryos were produced by culturing 15-20 COCs in BO capacitated sperms from frozen and thawed buffalo semen. SCNT embryo production rate was lower than IVF. IGF-1 mRNA expression was significantly upregulated at 2-cell, 16-cell, morula and blastocyst stages of SCNT embryos than in IVF embryos. IGF-1R expression declined from the 2-cell to the 16-cell stage, which increased in later stages and was highest in IVF blastocysts. Similar regulation was observed at different stages in SCNT embryos, except at the 4-cell stage where expression was higher. IGF-2 expression decreased up to the 8-cell stage and increased until the blastocyst stage, being higher in SCNT than IVF embryos. IGF-2R mRNA transcript expression was consistently lower in SCNT than in IVF embryos. Reprogramming of IGF-1, IGF-1R, IGF-2 and IGF-2R expression may have a significant role in cell proliferation in cloned embryos and is developmentally regulated.
Pandey
Alok
A
Transgenic Research Laboratory, DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, NH-1, Baldi By Pass, Karnal, Haryana, India.
Singh
Namita
N
Gupta
S C
SC
Rana
J S
JS
Gupta
Neelam
N
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002020:Buffaloes / Q000196:embryology / Q000235:genetics / Q000378:metabolism; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D007335:Insulin-Like Growth Factor II / Q000235:genetics / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D017526:Receptor, IGF Type 1; D017527:Receptor, IGF Type 2
D017527:Receptor, IGF Type 2; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D017526:Receptor, IGF Type 1