Significant Improvement in Cloning Efficiency of an Inbred Miniature Pig by Histone Deacetylase Inhibitor Treatment after Somatic Cell Nuclear Transfer1

After somatic cell nuclear transfer (SCNT), the epigenetic state of a differentiated donor cell nucleus must be reversed to the embryonic state. Incomplete epigenetic reprogramming and abnormal gene activation of the donor cell nuclei is thought to be the cause of low cloning efficiency. To improve cloning efficiency, we investigated the effect of scriptaid, a novel histone deacetylase inhibitor, on the in vitro development of porcine SCNT embryos were investigated. Cumulus cells collected from cumulus-oocyte complexes (COC) after 44 h of maturation were used for donor cell, and embryos were cultured in porcine zygote medium (PZM)-5 medium for 7 days. We found that treating SCNT embryos with 300 or 500 nM scriptaid for 20 h after activation increased developmental rate to the blastocyst stage (300 nM, 26.2%; 500 nM, 24.6% v. 100 nM, 18.3%; Ctrl, 15.7%; P < 0.05) and total cell numbers (300 nM, 43.5; 500 nM, 40.8 v. 100 nM, 33.8; Ctrl, 32.3; P < 0.05). Additionally, results of the TUNEL assay indicated that scriptaid decreased apoptosis (300 nM, 6.8% v. Ctrl, 11.4%; P < 0.05) in SCNT blastocysts. After the 300 nM scriptaid treatment, the levels of acetylated histone H3 lysine 9 and 5-hydroxymethylcytosines were increased (P < 0.05), and histone H3 lysine 9 trimethylation and 5-methylcytosine were decreased at the 1-cell stage, which might explain the enhanced (P < 0.05) transcript levels of mir-152, Oct4, Cdx2, and Bcl-xL and reduced (P < 0.05) transcription of Dnmt1, Casp3, and Bax in blastocysts. In conclusion, scriptaid enhances the developmental capacity by preventing apoptosis, and improves nuclear reprogramming in porcine SCNT embryos.This work was supported by the Bio-industry Technology Development Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea, and by a grant from the Next-Generation BioGreen 21 Program (No. PJ009601 and PJ009098), Rural Development Administration, Republic of Korea.

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Gene expression-signature of belinostat in cell lines is specific for histone deacetylase inhibitor treatment, with a corresponding signature in xenografts

Anti-Cancer Drugs ◽

10.1097/cad.0b013e32832e14e1 ◽

2009 ◽

Vol 20 (8) ◽

pp. 682-692 ◽

Cited By ~ 16

Author(s):

Anne Monks ◽

Curtis D. Hose ◽

Patrick Pezzoli ◽

Sudhir Kondapaka ◽

Gordon Vansant ◽

...

Keyword(s):

Gene Expression ◽

Histone Deacetylase ◽

Cell Lines ◽

Histone Deacetylase Inhibitor ◽

Gene Expression Signature ◽

Expression Signature ◽

Deacetylase Inhibitor ◽

Inhibitor Treatment

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Real-time monitoring of deformed wing virus-infected bee foraging behavior following histone deacetylase inhibitor treatment

iScience ◽

10.1016/j.isci.2021.103056 ◽

2021 ◽

pp. 103056

Author(s):

Cheng-Kang Tang ◽

Yu-Hsien Lin ◽

Joe-Air Jiang ◽

Yun-Heng Lu ◽

Chih-Hsuan Tsai ◽

...

Keyword(s):

Real Time ◽

Histone Deacetylase ◽

Foraging Behavior ◽

Histone Deacetylase Inhibitor ◽

Real Time Monitoring ◽

Deformed Wing Virus ◽

Deacetylase Inhibitor ◽

Bee Foraging ◽

Inhibitor Treatment

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Histone deacetylase inhibitor treatment induces ‘BRCAness’ and synergistic lethality with PARP inhibitor and cisplatin against human triple negative breast cancer cells

Oncotarget ◽

10.18632/oncotarget.2154 ◽

2014 ◽

Vol 5 (14) ◽

pp. 5637-5650 ◽

Cited By ~ 84

Author(s):

Kyungsoo Ha ◽

Warren Fiskus ◽

Dong Soon Choi ◽

Srividya Bhaskara ◽

Leandro Cerchietti ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Histone Deacetylase ◽

Triple Negative Breast Cancer ◽

Histone Deacetylase Inhibitor ◽

Breast Cancer Cells ◽

Triple Negative ◽

Parp Inhibitor ◽

Deacetylase Inhibitor ◽

Inhibitor Treatment

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41 DEVELOPMENT OF SOMATIC CELL NUCLEAR TRANSFERRED RAT EMBRYOS TO THE BLASTOCYST BY A HISTONE DEACETYLASE INHIBITOR, TRICHOSTATIN A

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab41 ◽

2011 ◽

Vol 23 (1) ◽

pp. 126

Author(s):

N. Kashiwazaki ◽

N. Nakajima ◽

K. Fujimaki ◽

K. Syudo ◽

J. Ito

Keyword(s):

Somatic Cell ◽

Histone Deacetylase ◽

Histone Deacetylase Inhibitor ◽

Trichostatin A ◽

Cell Stage ◽

Rat Embryos ◽

Reconstructed Embryos ◽

Deacetylase Inhibitor ◽

Histone Deacetylase Inhibitor Trichostatin ◽

Pronuclear Formation

Although cloned animals have been reproduced in many species via somatic cell nuclear transfer (SCNT), only 1 previous report showed successful generation of cloned rats. Some researchers evaluated developmental competence of reconstructed rat embryos and showed that these reconstructed embryos were never progressed beyond the 2-cell stage because of inadequate formation of the mitotic assembly and nuclear organisation or improper transcription of cytoskeleton genes. Even though our group also improved the proportion of 2 pronuclear formation in rat reconstructed embryos by the treatment with a proteasome inhibitor, MG132 (Nakajima et al. 2008 Cloning and Stem Cells 10, 461–468), no embryos were developed beyond the 2-cell stage. Recently, it has been demonstrated that a histone deacetylase inhibitor, trichostatin A (TSA) treatment dramatically improves efficiency of cloning in the mouse, although the precise effect of TSA is unclear in SCNT. However, in the case of rats, availability of TSA has not been tested. The aim of the present study was to clarify whether TSA treatment was also effective in rat SCNT. According to our previous reports, rat oocytes were collected and the spindles of the oocytes were removed in medium supplemented with MG132. After enucleation, nuclei of cumulus cells were injected in the oocytes and the reconstructed embryos were cultured in the medium supplemented with 0 (control), 5, 50, or 500 nM TSA. More than 100 embryos are used for each treatment group. Pronuclear formation, development to the 2-cell stage and blastocyst formation were evaluated. Our results demonstrated that TSA treatment affected neither the proportion of pronuclear formation nor development to the 2-cell stage in rat reconstructed embryos. However, the treatment with higher concentrations of TSA treatment (50 nM and 500 nM) compared to the concentration (5 nM) which was usually used for mouse SCNT enabled the reconstructed embryos to develop to the blastocyst stage but at low rate (1.4 to 2.2%). In both the 5 nM treatment and control groups, none of the rat reconstructed embryos developed to blastocyst. Taken together, our data suggests that TSA treatment also seems to be effective in rat SCNT. These findings will be useful for improvement of the protocol in rat SCNT. This work was also supported in part by the Promotion and Mutual Aid Corporation for Private Schools of Japan, Grant-in-Aid for Matching Fund Subsidy for Private Universities to J. I. and N. K.

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