Changes in acetylation of lysine 5 on histone H4 in canine oocytes following in vitro maturation

Changes in histone H4 acetylation during in vivo versus in vitro maturation of equine oocytes

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gar077 ◽

2011 ◽

Vol 18 (5) ◽

pp. 243-252 ◽

Cited By ~ 35

Author(s):

Federica Franciosi ◽

Valentina Lodde ◽

Ghylène Goudet ◽

Guy Duchamp ◽

Stefan Deleuze ◽

...

Keyword(s):

In Vitro Maturation ◽

Histone H4 ◽

Histone H4 Acetylation

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In vitro maturation affects chromosome segregation, spindle morphology and acetylation of lysine 16 on histone H4 in horse oocytes

Reproduction Fertility and Development ◽

10.1071/rd15350 ◽

2017 ◽

Vol 29 (4) ◽

pp. 721 ◽

Cited By ~ 8

Author(s):

Federica Franciosi ◽

Ghylene Goudet ◽

Irene Tessaro ◽

Pascal Papillier ◽

Rozenn Dalbies-Tran ◽

...

Keyword(s):

Chromosome Segregation ◽

Histone Deacetylases ◽

In Vitro Maturation ◽

Sirtuin 1 ◽

Histone H4 ◽

Histone Deacetylase 1 ◽

Dependent Protein ◽

H4k16 Acetylation ◽

Ovulatory Follicles

Implantation failure and genetic developmental disabilities in mammals are caused by errors in chromosome segregation originating mainly in the oocyte during meiosis I. Some conditions, like maternal ageing or in vitro maturation (IVM), increase the incidence of oocyte aneuploidy. Here oocytes from adult mares were used to investigate oocyte maturation in a monovulatory species. Experiments were conducted to compare: (1) the incidence of aneuploidy, (2) the morphology of the spindle, (3) the acetylation of lysine 16 on histone H4 (H4K16) and (4) the relative amount of histone acetyltransferase 1 (HAT1), K(lysine) acetyltransferase 8 (KAT8, also known as MYST1), histone deacetylase 1 (HDAC1) and NAD-dependent protein deacetylase sirtuin 1 (SIRT1) mRNA in metaphase II stage oocytes that were in vitro matured or collected from peri-ovulatory follicles. The frequency of aneuploidy and anomalies in spindle morphology was increased following IVM, along with a decrease in H4K16 acetylation that was in agreement with our previous observations. However, differences in the amount of the transcripts investigated were not detected. These results suggest that the degradation of transcripts encoding for histone deacetylases and acetyltransferases is not involved in the changes of H4K16 acetylation observed following IVM, while translational or post-translational mechanisms might have a role. Our study also suggests that epigenetic instabilities introduced by IVM may affect the oocyte and embryo genetic stability.

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210 POST-TRANSLATIONAL MODIFICATION OF HISTONE H4 ACETYLATION DURING IN VITRO MATURATION OF CANINE OOCYTES

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab210 ◽

2016 ◽

Vol 28 (2) ◽

pp. 236 ◽

Cited By ~ 1

Author(s):

T. F. Motheo ◽

D. R. Arnold ◽

W. R. R. Vicente ◽

B. I. Macente ◽

F. G. F. Filgueira ◽

...

Keyword(s):

In Vitro Maturation ◽

Least Square ◽

Penicillin G ◽

In Vitro Fertilisation ◽

Histone H4 ◽

Post Translational Modification ◽

Germinal Vesicle Breakdown ◽

Meiotic Process ◽

Histone H4 Acetylation

Proper in vitro oocyte maturation, crucial for in vitro fertilisation, is poorly understood and inefficient in dogs. Post-translational modifications of histones are involved in the meiotic process and preparation of oocytes for fertilisation. The present study aimed to evaluate post-translational histone H4 acetylation at lysine 12 (H4K12ac) and 16 (H4K16ac) in canine oocytes during in vitro maturation (IVM). Ovaries were retrieved from 1–7 year-old bitches undergoing routine ovariohysterectomy (OH). Grade I cumulus-oocyte complexes (COC) were recovered and evaluated before and after IVM in TCM 199 + 0.025 mM sodium pyruvate + 100 000 IU mL–1 penicillin G sodium + 100 mg mL–1 streptomycin sulfate, 0.003 g mL–1 BSA + 50 μL mL–1 Pluset® (500 UI FSH + 500 UI LH) + 1 μL mL–1 insulin-like growth factor (IGF) at 38.5°C in 5% CO2 for 72 h. After IVM, the nuclear stage of oocytes was established using Hoechst 33342 staining, and the acetylation patterns were investigated by indirect immunofluorescence staining of fixed immature and post-IVM oocytes (4% paraformaldehyde) with specific antibodies against the acetylated lysine residues, H4K12ac and H4K16ac. All stages were collected and evaluated simultaneously, and the experiment was repeated 4 times, with a total of 5–14 of oocytes evaluated per stage. Immunofluorescence signal was quantified using the NIH ImageJ software 1.4, and data are presented as a percentage of the average fluorescence intensity of each specific antibody over the intensity of DNA, as determined by Hoescht staining. Immunofluorescence values were analysed using the least square ANOVA by the general linear model procedures of SAS (SAS Institute, Cary, NC, USA), and comparisons of means were further performed by Fisher’s Exact test. A probability level of P < 0.05 was considered significant. Our results indicate a variation in acetylation levels of H4K12 (P < 0.02) and H4K16 (P < 0.04) during meiosis. Fluorescence levels of H4K12ac decreased significantly when comparing immature (2.19 ± 0.62) with oocytes in metaphase I/II (MI/MII) (0.52 ± 0.14). Similarly, H4K16ac displayed a greater acetylation pattern in immature oocytes (1.20 ± 0.27) when compared to oocytes in germinal vesicle breakdown (GVBD; 0.58 ± 0.16) and MI/MII (0.62 ± 0.07) stages. These results support the hypothesis that, similar to other domestic species, post-translational modification of histone acetylation takes place during the meiotic process of oocyte IVM. Whether these histone modifications take place in a similar fashion during IVM remains to be investigated.

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