NDA@SZTAKI

A comparison of alternative assays to measure DNA damage in stallion spermatozoa: TUNEL test versus ‘Nicoletti assay’

Kakasi, Balázs

Nagy, Szabolcs

Pál, László

Czimber, Gyula

Husvéth, Ferenc

Akadémiai Kiadó

2015

SV Veterinary science / állatorvostudomány

The aberrations of sperm DNA may cause various problems and have negative consequences on fertility. These influence embryonic development or might lead to early embryo loss. Sperm Chromatin Structure Assay (SCSA) is the flow cytometric method most often used for the detection of DNA lesions; however, some studies using that method reached confusing conclusions. The aim of this pilot study was to adjust and compare two alternative tests, namely the TUNEL test and the Nicoletti assay. The above-mentioned two flow cytometric methods capable of detecting the fragmented DNA of sperm were tested on 12 frozen-thawed stallion semen samples. The TUNEL test demonstrated much higher DNA fragmentation ratio than the Nicoletti assay (mean ± SD: 30.77 ± 13.03% vs. 1.93 ± 0.89%, respectively). A fluorescent microscopic check of the samples showed that TUNEL labelled the plasma membrane and the mitochondria in a nonspecific way, rather than detecting only the fragmented DNA, thus eventually resulting in a false positive sign. The Nicoletti assay is simpler, quicker and does not detect nonspecific binding; however, further analyses are required to determine its diagnostic value.

angol

Article

PeerReviewed

info:eu-repo/semantics/article

text

http://real.mtak.hu/36070/1/avet.2015.010.pdf

Kakasi, Balázs and Nagy, Szabolcs and Pál, László and Czimber, Gyula and Husvéth, Ferenc (2015) A comparison of alternative assays to measure DNA damage in stallion spermatozoa: TUNEL test versus ‘Nicoletti assay’. Acta Veterinaria Hungarica, 63 (1). pp. 118-124. ISSN 0236-6290

http://real.mtak.hu/36070/

http://dx.doi.org/10.1556/AVet.2015.010