Investigation and Assessment of Genome Integrity in Mitochondria of Testis Tissue, Cell Proliferation and Topoisomerase Assay in invitro Condition

Aim: Assessment of genome integrity in testis and to explain the reasons for infertility with the drug
metosartan.
Methodology: Comet assay with freshly isolated mitochondria, cryopreserved mitochondria and
cryopreserved tissue mitochondria and cell proliferation assay by dye exclusion method and
topoisomerase assay in fresh and deep freezed DNA.
Results and Discussion: Comet assay of isolated mitochondria both from fresh and cryopreserved
samples was examined by fluorescence microscope, which revealed that cryopreservation of testes
mitochondria samples lead to injury to the tissue and DNA fragmentation. Topo isomerase assay has
showed that increased incubation leads to DNA fragmentation and cell proliferation assay with the
drug has showed the 0% viability of the cells by trypan blue. So, in vivo studies are necessary to
confirm whether the drug is safe or not.
Conclusion: Assessment of genome integrity lead to the finding that drug was not safe it resulted in
so many difficulties in invitro condition. The drug metosartan resulted in formation of micronuclei in
mitochondrial genome and also decreased viability by trypan blue, topoisomerase assay has revealed
that with incubation the DNA was fragmented resulting in the bands as supercoiled and fragmented
DNA.