Epigenetic modification of line 1 and hervk-10 methylation in gametes of ifertile males (Doctoral thesis)

Κωστούλας, Χαρίλαος Α.

We intend to investigate the overall methylation level and the four different patterns on methylation status of two CpGs (mCmC, mCuC, uCmC, uCuC) of LINE-1 on normozoospermic and oligospermic men. DNA methylation in mammals is a dynamic and strictly controlled process which is involved in numerous cellular processes, cell differentiation, regulation of gene expression, genome reprogramming and silencing of repetitive elements. In mammals, the epigenetic reprogramming is crucial for germ cells development. The principal epigenetic regulatory mechanism is DNA methylation and occurs in the phase of gametogenesis and upon early embryo stages. During gametogenesis there are occurred periods of DNA methylation and demethylation. There are 500.00 copies of LINE-1 in the human genome. Aberrant DNA methylation levels of LINE-1 have been implicated in various human diseases. The study consisted of 92 men. A detailed medical history was obtained from all subjects. The subjects were split into two groups: normozoospermic (70 men) and oligospermic (22 men, <20x106 spermatozoa). The oligospermic group was classified into Mid (1 man, 10-20x106 spermatozoa), Moderate (12 men, 5-10x106 spermatozoa) and Severe (9 men <5x106 spermatozoa). Semen was analyzed according to the WHO criteria. LINE-1 overall methylation level and methylation pattern of the oligospermic group were compared to the normozoospermic one. Methylation level and pattern of LINE-1 was measured via COBRA. DNA was extracted from human sperm and treated with sodium Bisulfite. DNA was amplified using primers which were designed to amplify 5΄UTR. COBRALINE-1 results were sorted into four groups according on the methylation the status of the two CpG dinucleotides from the 5΄-3΄ends of sequence: hypermethylation (mCmC), hypomethylation (uCuC), partial methylation of the 5΄(mCuC) and partial methylation of the 3΄(uCmC). Our data show a difference in the overall methylation level and different correlation in the methylation pattern on normozoospermic and oligospermic men. Our findings show that in normozoospemic samples the unmethylated sequences derived from partially methylated LINE-1s mCuC motif. Unlike, in oligospermic samples the inverse correlation indicates change of methylated LINE-1 with partially mCuC pattern. Therfore, the difference in methylation levels show different genomic allocation of LINE-1 methylation between normozoospemic and oligospermic.
Institution and School/Department of submitter: Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής
Subject classification: Σπερματοζωάρια
Keywords: Επιγενετική,Γονιδιακή αποτύπωση,Μεθυλίωση,Ρετρομεταθετά,Σπερματοζωάρια,Μοτίβα,Ρετρομετάθεση,Διαφορικά μεθυλιωμένες περιοχές,Epigenetics,Genomic imprinting,Methylation,Retroelements,Spermatozoa,Motifs,Differentially methylated regions
URI: https://olympias.lib.uoi.gr/jspui/handle/123456789/29361
Appears in Collections:Διδακτορικές Διατριβές

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