Science and Technology Production
Article
Authorship
Monachesi, Norma
;
Neild, Deborah
;
CARRETERO, MARIA IGNACIA
Date
2019
Publishing House and Editing Place
WILEY-BLACKWELL PUBLISHING, INC
Magazine
REPRODUCTION IN DOMESTIC ANIMALS (1990),
vol. 54
(pp. 1078-1084)
WILEY-BLACKWELL PUBLISHING, INC
Summary
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SIGEVA
The toluidine blue (TB) stain has been used in different species to evaluate the degree of chromatin condensation. The objectives of this study were: simplify the TB stain to evaluate sperm in canine raw semen, verify the staining patterns for this species using this simplified technique and establish a protocol for using dithiotreitol (DTT) as a positive control for TB staining in dogs. Twenty-one ejaculates were collected from 7 adult male dogs; semen was extended, fixed with ethanol 96° ...
The toluidine blue (TB) stain has been used in different species to evaluate the degree of chromatin condensation. The objectives of this study were: simplify the TB stain to evaluate sperm in canine raw semen, verify the staining patterns for this species using this simplified technique and establish a protocol for using dithiotreitol (DTT) as a positive control for TB staining in dogs. Twenty-one ejaculates were collected from 7 adult male dogs; semen was extended, fixed with ethanol 96° and stained with TB using 2 staining times: 15 and 30 minutes. In addition, 3 incubation times with 1% DTT were assayed (2, 5 and 30 minutes). Three staining patterns were established: light blue coloring (TB negative, normal chromatin condensation), light violet (TB intermediate, some degree of chromatin decondensation) and dark blue-violet (TB positive, high degree of chromatin decondensation). No significant differences (p>0.05) were observed between the staining times (15 and 30 minutes) for any of the TB patterns. All DTT incubation times (2, 5 and 30 minutes) showed 100% sperm positive to TB. To conclude, it was possible to simplify the TB stain and determine the different patterns in canine spermatozoa. Also, DTT can be used both as a positive control for the stain and to evaluate individual susceptibility to decondensation in vitro. This article is protected by copyright. All rights reserved.
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Key Words
DNATOLUIDINE BLUESPERMDOG