Buffalo (Bubalus bubalis) SCNT embryos produced from somatic cells isolated from frozen-thawed semen: effect of trichostatin A on the in vitro and in vivo developmental potential, quality and epigenetic status.
This study examined the effects of trichostatin A (TSA) treatment of reconstructed buffalo embryos, produced by hand-made cloning using somatic cells isolated from over a decade old frozen-thawed semen, on their in vitro and in vivo developmental competence, quality and epigenetic status. Following treatment of reconstructed embryos with TSA (0, 50 or 75 nM) for 10 h prior to culture, the cleavage (100.0 ± 0, 94.5 ± 2.3 and 96.1 ± 1.2%, respectively) and blastocyst rate (50.6 ± 2.3, 48.4 ± 2.7 and 48.1 ± 2.6%, respectively), total cell number (275 ± 17.4, 289 ± 30.1 and 317 ± 24.2, respectively) and apoptotic index (5.6 ± 0.7, 3.4 ± 0.9 and 4.5 ± 1.4, respectively) were not significantly different among the three groups. However, TSA treatment increased (P < 0.05) the global level of H4K5ac and decreased (P < 0.05) that of H3K27me3 in blastocysts whereas the global level of H3K18ac was not affected significantly. Transfer of embryos treated with 75 nM TSA (n = 10) to recipients resulted in two pregnancies (20%), one out of which was aborted in the second and the other in the third trimester whereas transfer of control embryos (n = 20) or those treated with 50 nM TSA (n = 12) did not result in any pregnancy. In conclusion, these results suggest that TSA treatment of cloned buffalo embryos produced using somatic cells isolated from frozen-thawed semen improved their epigenetic status but not the in vitro developmental potential and offspring rate.
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Naresh L
NL
Selokar
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Monika
M
Saini
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Himanshu
H
Agrawal
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Prabhat
P
Palta
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Manmohan S
MS
Chauhan
Animal Biotechnology Centre,National Dairy Research Institute,Karnal 132001,India.
Radheysham
R
Manik
Animal Biotechnology Centre,National Dairy Research Institute,Karnal,India.
Suresh K
SK
Singla
D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002020:Buffaloes; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects* / Q000235:genetics; D044127:Epigenesis, Genetic / Q000187:drug effects* / Q000235:genetics; D005260:Female; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D008745:Methylation / Q000187:drug effects; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D018873:Pregnancy Rate; D012661:Semen / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012662:Semen Preservation
D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; C012589:trichostatin A
Buffalo; Embryo; Hand-made cloning; SCNT; Trichostatin A