title: Changes in acetylation of lysine 5 on histone H4 in canine oocytes following in vitro maturation.
abstract: Post-translational modifications of histones, such as acetylation, are involved in regulating chromatin remodelling and gene expression. Proper in vitro maturation (IVM) of canine oocytes, for many reasons, is up to now inefficient. This study aimed to evaluate the post-translational histone H4 acetylation at lysine 5 (H4K5) in immature and post-IVM canine oocytes. Oocyte nuclear stage was assessed using Hoechst 33342 staining. Acetylation patterns were determined by indirect immunofluorescence staining of immature and post-IVM oocytes, using an antibody against the acetylated lysine 5 residue on histone 4 (H4K5ac). The experiment was repeated four times, with a total of 7-17 oocytes evaluated per stage. Immunofluorescence signal was quantified using the NIHimagej software. Data were expressed as a percentage of the average fluorescence intensity of the specific antibody over the intensity of DNA, as determined by Hoescht staining. H4K5ac displayed a significantly higher acetylated pattern in immature oocytes (0.97 ± 0.08) when compared to post-IVM oocytes at different nuclear stages. There was a decrease in the fluorescence level of the matured oocytes with the progression of meiosis (GVBD: 0.47 ± 0.06 and MI/MII: 0.35 ± 0.04). Similarly to other domestic species, we hypothesized that post-translational modification of histone acetylation takes place during meiosis of in vitro matured canine oocytes. However, it remains to be investigated whether these changes occur during in vivo maturation.
authors.affiliation: Department of Animal Reproduction, Faculdade de Ciencias Agrarias e Veterinarias Campus de Jaboticabal, Universidade Estadual Paulista Julio de Mesquita Filho, Jaboticabal, Brazil.
authors.forename: T F
authors.initials: TF
authors.lastname: Motheo
authors.affiliation: In vitro Brasil S/A, Mogi Mirim, Brazil.
authors.forename: D R
authors.initials: DR
authors.lastname: Arnold
authors.affiliation: Department of Animal Reproduction, Faculdade de Ciencias Agrarias e Veterinarias Campus de Jaboticabal, Universidade Estadual Paulista Julio de Mesquita Filho, Jaboticabal, Brazil.
authors.forename: L C
authors.initials: LC
authors.lastname: Padilha-Nakaghi
authors.affiliation: Department of Animal Reproduction, Faculdade de Ciencias Agrarias e Veterinarias Campus de Jaboticabal, Universidade Estadual Paulista Julio de Mesquita Filho, Jaboticabal, Brazil.
authors.forename: E A
authors.initials: EA
authors.lastname: Pires-Buttler
authors.affiliation: Universidade Federal de Uberlandia - Campus Umuarama, Uberlandia, Brazil.
authors.forename: A E
authors.initials: AE
authors.lastname: Alves
authors.affiliation: Programa de Mestrado em Ciência Animal, Universidade de Franca, Franca, Brazil.
authors.forename: M
authors.initials: M
authors.lastname: Apparicio
authors.affiliation: Department of Animal Reproduction, Faculdade de Ciencias Agrarias e Veterinarias Campus de Jaboticabal, Universidade Estadual Paulista Julio de Mesquita Filho, Jaboticabal, Brazil.
authors.forename: Wrr
authors.initials: W
authors.lastname: Vicente
authors.affiliation: Department of Support, Production and Animal Health, Faculdade de Medicina Veterinária de Araçatuba - FMVA/UNESP, Universidade Estadual Júlio de Mesquita Filho, Araçatuba, Brazil.
authors.forename: F L
authors.initials: FL
authors.lastname: Lopes
mesh_terms: D000107:Acetylation; D000818:Animals; D004285:Dogs / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro; D019084:Fluorescent Antibody Technique, Indirect; D006657:Histones / Q000737:chemistry*; D059471:In Vitro Oocyte Maturation Techniques; D008540:Meiosis / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D009866:Oogenesis / Q000502:physiology*
chemical_list: D006657:Histones