In situ viability detection assays induce heat-shock protein 70 expression in spermatozoa without affecting the chromatin integrity

Asokan, Y and Honguntikar, Sachin D and Uppangala, S and Salian, SR and Kumar, D and Kalthur, Guruprasad and Adiga, Satish Kumar (2014) In situ viability detection assays induce heat-shock protein 70 expression in spermatozoa without affecting the chromatin integrity. Andrologia, 20. pp. 1-8. ISSN 0303-4569

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Abstract

To differentiate dead spermatozoa from viable but immotile spermatozoa, several techniques are being used during ICSI. As processed spermatozoa from poor-quality ejaculate are confronted with a higher risk of experiencing stress on exposure to altered osmotic conditions or chemicals, this study was undertaken to determine the expression of stress response gene Hsp70 and chromatin integrity in spermatozoa subjected to in situ viability assays such as hypo-osmotic swelling (HOS) test, modified hypo-osmotic swelling (M-HOS) test and pentoxifylline in 25 fresh and frozen–thawed asthenozoospermic ejaculates. RT-PCR and immunofluorescence detection of Hsp70 were performed to elucidate the expression and localisation of Hsp70 in spermatozoa, whereas DNA fragmentation analysis was performed by sperm chromatin dispersion assay. Exposure of fresh and frozen–thawed asthenozoospermic spermatozoa to M-HOS and pentoxifylline significantly increased Hsp70 expression as evidenced by increased RNA expression and immunolocalisation of Hsp70 protein in sperm head (P < 0.05–0.001). However, chromatin integrity was not significantly affected in any groups until 6 h of post-exposure time period. Our results suggest that conventional HOS may be preferred for the in situ detection of the viability as there was no immediate stress response and chromatin instability in the exposed spermatozoa.

Item Type: Article
Uncontrolled Keywords: Hsp70; hypo-osmotic swelling test ; pentoxifylline; sperm chromatin integrity; sperm viability.
Subjects: Medicine > KMC Manipal > Obstetrics & Gynaecology
Depositing User: KMC Manipal
Date Deposited: 23 Oct 2014 06:25
Last Modified: 23 Oct 2014 06:25
URI: http://eprints.manipal.edu/id/eprint/140911

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