High-fat diet-induced and genetically inherited obesity differentially alters DNA methylation profile in the germline of adult male rats.

Clinical Epigenetics
Sharvari S DeshpandeNafisa H Balasinor

Abstract

Paternal obesity has been associated with reduced live birth rates. It could lead to inheritance of metabolic disturbances to the offspring through epigenetic mechanisms. However, obesity is a multifactorial disorder with genetic or environmental causes. Earlier we had demonstrated differential effects of high-fat diet-induced obesity (DIO) and genetically inherited obesity (GIO) on metabolic, hormonal profile, male fertility, and spermatogenesis using two rat models. The present study aimed to understand the effect of DIO and GIO on DNA methylation in male germline, and its subsequent effects on the resorbed (post-implantation embryo loss) and normal embryos. First, we assessed the DNA methylation enzymatic machinery in the testis by Real-Time PCR, followed global DNA methylation levels in spermatozoa and testicular cells by ELISA and flow cytometry, respectively. Further, we performed Methylation Sequencing in spermatozoa for both the groups. Sequencing data in spermatozoa from both the groups were validated using Pyrosequencing. Expression of the differentially methylated genes was assessed in the resorbed and normal embryos sired by the DIO group using Real-Time PCR for functional validation. We noted a significant decrease...Continue Reading

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Methods Mentioned

BETA
flow cytometry
ELISA
methylome sequencing
methylation sequencing
Methyl Seq
FACS
Methyl-Seq
PCR

Software Mentioned

FACS Diva
Bowtie2
PERL script
Graph Pad Prism
Bismarkv0
MIQE
FastQC
PANTHER
DESeq
build

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