open access

Vol 47, No 4 (2009)
Original paper
Submitted: 2011-12-19
Published online: 2010-05-01
Get Citation

Modification of equine sperm chromatin decondensation method to use fluorescence in situ hybridization (FISH).

Monika Bugno-Poniewierska, Zofia Jabłońska, Ewa Słota
DOI: 10.2478/v10042-010-0006-2
·
Folia Histochem Cytobiol 2009;47(4):663-666.

open access

Vol 47, No 4 (2009)
ORIGINAL PAPERS
Submitted: 2011-12-19
Published online: 2010-05-01

Abstract

Fluorescence in situ hybridization (FISH) is widely used in the study of chromosome structure and organization. Cytogenetic evaluation of chromosomes using FISH technique plays an increasingly important role in diagnosing karyotype changes in both somatic and reproductive cells. The aim of the study was to optimize the conditions of stallion sperm decondensation, which have a significant effect on the results of fluorescence in situ hybridization. Appropriate type and time of decondensation was chosen for the sperm of every stallion. It was found that decondensation performed using a preparation incubated in DTT solution for 1.5 minutes and in SDS solution for 10 seconds proved effective for stallions no. 1 and 2. An alternative decondensation method performed in an Eppendorf tube, with incubation in DTT solution for 1 minute and in SDS solution for 5 seconds proved effective for stallions no. 3 and 4. Decondensation using DTT and papain solution, a method successfully used for bull spermatozoa, proved inadequate for horse spermatozoa.

Abstract

Fluorescence in situ hybridization (FISH) is widely used in the study of chromosome structure and organization. Cytogenetic evaluation of chromosomes using FISH technique plays an increasingly important role in diagnosing karyotype changes in both somatic and reproductive cells. The aim of the study was to optimize the conditions of stallion sperm decondensation, which have a significant effect on the results of fluorescence in situ hybridization. Appropriate type and time of decondensation was chosen for the sperm of every stallion. It was found that decondensation performed using a preparation incubated in DTT solution for 1.5 minutes and in SDS solution for 10 seconds proved effective for stallions no. 1 and 2. An alternative decondensation method performed in an Eppendorf tube, with incubation in DTT solution for 1 minute and in SDS solution for 5 seconds proved effective for stallions no. 3 and 4. Decondensation using DTT and papain solution, a method successfully used for bull spermatozoa, proved inadequate for horse spermatozoa.
Get Citation
About this article
Title

Modification of equine sperm chromatin decondensation method to use fluorescence in situ hybridization (FISH).

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 47, No 4 (2009)

Article type

Original paper

Pages

663-666

Published online

2010-05-01

DOI

10.2478/v10042-010-0006-2

Bibliographic record

Folia Histochem Cytobiol 2009;47(4):663-666.

Authors

Monika Bugno-Poniewierska
Zofia Jabłońska
Ewa Słota

Regulations

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By "Via Medica sp. z o.o." sp.k., ul. Świętokrzyska 73, 80–180 Gdańsk

tel.:+48 58 320 94 94, faks:+48 58 320 94 60, e-mail:  viamedica@viamedica.pl