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2017 ; 63
(4
): 365-375
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Epigenetic analysis of bovine parthenogenetic embryonic fibroblasts
#MMPMID28484201
Kaneda M
; Takahashi M
; Yamanaka KI
; Saito K
; Taniguchi M
; Akagi S
; Watanabe S
; Nagai T
J Reprod Dev
2017[Aug]; 63
(4
): 365-375
PMID28484201
show ga
Although more than 100 imprinted genes have already been identified in the mouse
and human genomes, little is known about genomic imprinting in cattle. For a
better understanding of these genes in cattle, parthenogenetically activated
bovine blastocysts were transferred to recipient cows to obtain parthenotes, and
fibroblasts derived from a Day 40 (Day 0 being the day of parthenogenetic
activation) parthenogenetic embryo (BpEFs) were successfully obtained. Bovine
embryonic fibroblasts (BEFs) were also isolated from a normal fertilized embryo
obtained from an artificially inseminated cow. The expression of imprinted genes
was analyzed by RT-PCR. Paternally expressed genes (PEGs) in mouse (viz., IGF2,
PEG3, ZAC1, NDN, DLK1, SGCE, and PEG10) were expressed in BEFs, but not in BpEFs,
suggesting that these genes are also imprinted in cattle. However, other PEGs in
mouse (viz., IMPACT, MAGEL2, SNRPN, and PEG1/MEST) were expressed in both BEFs
and BpEFs. These genes may not be imprinted in BEFs. The expression of seven
maternally expressed genes in mouse was also analyzed, and only CDKN1C was not
expressed in BpEFs. The DNA methylation patterns of repetitive elements
(Satellite I, Satellite II, alpha-satellite, and Art2) were not different between
the BEFs and BpEFs; however, the differentially methylated region (DMR) of
paternally methylated H19 was hypomethylated, whereas those of maternally
methylated PEG3 and PEG10 were hypermethylated in BpEFs, as expected. The
methylation of the SNRPN DMR was not different between the BEFs and BpEFs, in
accordance with the SNRPN expression levels in both cell types. The XIST gene,
which is essential for X chromosome inactivation in females, was expressed in
BpEFs, whereas its DMR was half-methylated, suggesting that X chromosome
inactivation is normal in these cells. Microarray analysis was also applied to
identify novel PEGs that should be expressed only in BEFs but not in BpEFs. More
than 300 PEG candidate genes, including IGF2, PEG3, and PEG10, were obtained.
These results illustrate the epigenetic characteristic of bovine parthenogenetic
embryos and contribute to the identification of novel imprinted genes in cattle.
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