Preliminary study of the difference in the genome-wide methylation profiles between germ cells and somatic cells of bulls

Phutikanit, N., Suwimonteerabutr, J., Harrison, D., D'Occhio, M. J., Carroll, B. J. and Techakumphu, M. (2007). Preliminary study of the difference in the genome-wide methylation profiles between germ cells and somatic cells of bulls. In: Reproduction, Fertility and Development: Proceedings of the Annual Conference of the International Embryo Transfer Society, Kyoto, Japan, 6-10 January 2007 Full Papers and Abstracts for Poster Presentation. International Embryo Transfer Society 2007 Meeting, Kyoto, Japan, (254-254). 6-10 January 2007. doi:10.1071/RDv19n1Ab277


Author Phutikanit, N.
Suwimonteerabutr, J.
Harrison, D.
D'Occhio, M. J.
Carroll, B. J.
Techakumphu, M.
Title of paper Preliminary study of the difference in the genome-wide methylation profiles between germ cells and somatic cells of bulls
Conference name International Embryo Transfer Society 2007 Meeting
Conference location Kyoto, Japan
Conference dates 6-10 January 2007
Proceedings title Reproduction, Fertility and Development: Proceedings of the Annual Conference of the International Embryo Transfer Society, Kyoto, Japan, 6-10 January 2007 Full Papers and Abstracts for Poster Presentation   Check publisher's open access policy
Journal name Reproduction Fertility and Development   Check publisher's open access policy
Place of Publication East Melbourne, Vic.
Publisher CSIRO in co-operation with the Australian Academy of Science
Publication Year 2007
Sub-type Published abstract
DOI 10.1071/RDv19n1Ab277
ISSN 1031-3613
Volume 19
Issue 1
Start page 254
End page 254
Total pages 1
Language eng
Abstract/Summary DNA methylation is tissue-specific and is thought to be one of the factors that regulates gene expression. This recent study was carried out in order to examine the difference of the genome-wide methylation profiles at the HpaII sites in germ cells and somatic cells of bulls. Ejaculated spermatozoa, leukocytes, and ear fibroblasts were collected from 3 Holstein bulls of ages 1 to 3 years. The genomic DNA was extracted and treated overnight with a methylation-sensitive restriction endonuclease (HpaII) to digest unmethylated sites throughout the genome. Both undigested and digested DNA samples were used as templates in the PCR-based technique developed by researchers at the University of Queensland, which allows the amplification of the methylation sites by short oligonucleotide primers of arbitrary sequence containing the HpaII recognition site (CCGG). The amplicons were separated in 4% polyacrylamide gel by electrophoresis and the gel was stained with silver nitrate. The results were evaluated on the basis of the presence–absence of the band(s) in the digested template compared with the undigested counterpart, and the difference between types of marker was analyzed using the chi-square test. From 10 sets of primer, approximately 400 markers in the genomic samples could be scored. The samples from the 3 bulls showed similar but not identical patterns. Statistical analysis showed that the difference between marker types was dependent on the individual. Generally, most of the markers were digestion-resistant markers signifying that most of the HpaII sites in the genome of both germ cells and somatic cells are methylated. Leukocytes had a significantly higher methylation content compared to fibroblasts (94.1 vs. 90.1%; P = 0.0004), but did not differ from those in sperm (92.3%; P = 0.09). Sperm cells showed a slightly higher percentage of unmethylated sites than did somatic cells (3.5 vs. 2.6% in leukocytes and 3.3% in fibroblasts), and yet the difference was non-significant. Moreover, fibroblastic cells had a higher portion of the digestion-dependent markers than did other cell types, and this difference was statistically significant (6.6 vs. 4.2% in sperm, P = 0.009, and 3.3% in leukocytes, P = 0.001). In conclusion, the DNA of the germ cells and somatic cells is highly methylated at the HpaII sites, with some variation in methylation pattern between the 2 cell lineages. The markers found only in the digested template of the ear fibroblasts suggest the difference in genome structure between the versatile cell types and the fully differentiated ones. Further investigation is required to elucidate any possible relationship between the variation of the methylation pattern found in sperm and fibroblasts and the failure of the reprogramming process in cloned animals derived from somatic cell nuclear transfer.
Subjects 07 Agricultural and Veterinary Sciences
Keyword Developmental Biology
Reproductive Biology
Zoology
Q-Index Code EX
Q-Index Status Provisional Code
Institutional Status Unknown

 
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Created: Mon, 18 Feb 2008, 15:55:45 EST