Methylation pattern and differential MHM region expression during chicken gonadal development

• Grant number: 09/08313-2
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2009
• End date: March 31, 2012
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Ester Silveira Ramos
• Grantee: Ester Silveira Ramos
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Birds present the ZZ/ZW sex chromosomal system (male and female, respectively). However, the mechanisms of sex determination are still unknown. It has been speculated that Z- and W-specific genes and the male hypermethylated (MHM) region on the Z chromosome are involved in the sex determination of birds. There are evidences that epigenetic mechanisms might be associated with the sex determination. The MHM region is transcribed in a non-coding RNA and accumulates at the site of transcription, similar to Xist RNA. The aim of the present study is to explore a model of sex determination and differentiation, and dosage compensation for research involving epigenetic aspects of gonads and germ cells. In the present study, we will investigate differential MHM region expression during chicken gonadal development and its methylation pattern. Embryos of Gallus gallus will be removed from eggs, staged according to the criteria of Hamburger and Hamilton, and their gonadal region will be excised prior to (20HH and 25HH), during (28HH and 35HH) and post-differentiation (40HH) of the gonad. Adult testis and ovary will also be analyzed. All samples will be sexed by CHD test. The methylation pattern will be analyzed by combined bisulfite-restriction analysis (COBRA) and methylation-sensitive enzymatic digestion associated to real time PCR. Total RNA will be extracted, reverse transcribed and the cDNA will be analyzed by real-time PCR. The MHM region transcripts will be detected and compared with the expression levels of the genes DMRT1, HINTZ, HINTW, AMH, DAX1, SOX3 and SOX9. The results may show a differential expression pattern and a correlation among the expression of these genes/region and the MHM region methylation pattern. These data are extremely important in the verification of the real MHM role in the gonadal development and in one possible and unique mechanism of dosage compensation. (AU)

Scientific publications

(8)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

H.R. MAGALHÃES; S.B.P. LEITE; C.C.P. DE PAZ; G. DUARTE; E.S. RAMOS. Placental hydroxymethylation vsmethylation at the imprinting control region 2 on chromosome 11p15.5. Brazilian Journal of Medical and Biological Research, v. 46, n. 11, p. 916-919, 2013-10-22. (09/08313-2, 10/03519-9)

CAETANO, L. C.; GENNARO, F. G. O.; COELHO, K.; ARAUJO, F. M.; VILA, R. A.; ARAUJO, A.; BERNARDO, A. DE MELO; MARCONDES, C. R.; LOPES, S. M. CHUVA DE SOUSA; RAMOS, E. S.. Differential expression of the MHM region and of sex-determining-related genes during gonadal development in chicken embryos. Genetics and Molecular Research, v. 13, n. 1, p. 838-849, 2014. (09/08313-2)

DA SILVA, A. M.; DE FREITAS, M. A. R.; RIOS, A. F. L.; RENZI, A.; LOBO, R. B.; GALERANI, M. A. V.; VILA, R. A.; RAMOS, E. S.. Identification of a DNA methylation point in the promoter region of the bovine CYP21 gene. Genetics and Molecular Research, v. 10, n. 3, p. 1409-1415, 2011. (09/08313-2)

LEMOS, D. C.; TAKEUCHI, P. L.; RIOS, A. F. L.; ARAUJO, A.; LEMOS, H. C.; RAMOS, E. S.. Bovine fetal DNA in the maternal circulation: Applications and implications. Placenta, v. 32, n. 11, p. 912-913, NOV 2011. (09/08313-2)

SOARES, M. R.; HUBER, J.; RIOS, A. F. L.; RAMOS, E. S.. Investigation of IGF2/ApaI and H19/RsaI polymorphisms in patients with cutaneous melanoma. GROWTH HORMONE & IGF RESEARCH, v. 20, n. 4, p. 295-297, AUG 2010. (07/01911-6, 09/08313-2)

MAGALHAES, H. R.; LEITE, S. B. P.; DE PAZ, C. C. P.; DUARTE, G.; RAMOS, E. S.. Placental hydroxymethylation vs methylation at the imprinting control region 2 on chromosome 11p15.5. Brazilian Journal of Medical and Biological Research, v. 46, n. 11, p. 4-pg., 2013-11-01. (10/03519-9, 09/08313-2)

EIRAS, MATHEUS CREDENDIO; MIRANDA-FURTADO, CRISTIANA LIBARDI; FERNANDES, ARTUR; RAMOS, ESTER SILVEIRA. A new chicken molecular sexing assay based on the Z chromosome dose and the MHM region. Theriogenology, v. 122, p. 84-87, DEC 2018. (09/08313-2)

CARNEIRO, M. C. A.; TAKEUCHI, P. L.; ARAUJO, A.; LOBO, R. B.; ELIAS, F. P.; VILA, R. A.; MIRANDA-FURTADO, C. L.; RAMOS, E. S.. Sexing single bovine blastomeres using TSPY gene amplification. Genetics and Molecular Research, v. 10, n. 4, p. 3937-3941, 2011. (09/08313-2)