| Full text | |
| Author(s): |
Rui, Bruno R.
[1]
;
Angrimani, Daniel S. R.
[1]
;
Losano, Joao Diego A.
[1]
;
Bicudo, Luana de Cassia
[1]
;
Nichi, Marcilio
[1]
;
Pereira, Ricardo J. G.
[1]
Total Authors: 6
|
| Affiliation: | [1] Univ Sao Paulo, Dept Anim Reprod, Coll Vet Med & Anim Sci, Ave Prof Orlando Marques de Paiva 87, BR-05508270 Sao Paulo - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | Animal Reproduction Science; v. 187, p. 133-140, DEC 2017. |
| Web of Science Citations: | 2 |
| Abstract | |
Several methods have been developed to evaluate spermatozoa function in birds but many of these are sometimes complicated, costly and not applicable to field studies (i.e., performed within poultry breeding facilities). The objective was, therefore, to validate efficient, practical and inexpensive procedures to determine DNA fragmentation, acrosomal integrity, and mitochondrial activity in poultry spermatozoa. Initially, ejaculates were individually diluted and divided into control (4 degrees C, 4 h) and UV-irradiated aliquots (room temperature, 4 h), and then samples containing different percentages of DNA-damaged spermatozoa (0%, 25%, 50%, 75% and 100%) were subjected to Toluidine Blue (TB) and Sperm Chromatin Dispersion assessments (SCD). Fast Green-Rose Bengal (FG-RB) and FITC-PSA staining protocols were subsequently used to assess acrosome status in aliquots comprising assorted amounts of acrosome-reacted spermatozoa. Furthermore, to validate 3,3'-diaminobenzidine (DAB) assay, ejaculates containing different gradients of spermatozoa with great amounts of mitochondrial activity were concurrently evaluated using DAB and JC-1 stains. The proportion of spermatozoa with abnormal DNA integrity when evaluated using the TB assessment correlated significantly with the expected percentages of UV-irradiated spermatozoa and with SCD results. A significant linear regression coefficient was also observed between expected amounts of acrosome-intact spermatozoa and FG-RB readings, and there was a significant correlation of the data when FG-RB and FITC-PSA were used. Likewise, the use of the DAB assay enabled for accurately ascertaining percentages of rooster spermatozoa with greater and lesser mitochondrial function, and results were highly correlated to results with staining with JC-1. Altogether, findings of the present study indicate acrosomal status, DNA integrity and mitochondrial activity in rooster spermatozoa can be easily and reliably determined using FG-RB, TB and DAB stains. (AU) | |
| FAPESP's process: | 12/08621-1 - Conservation of avian germplasm by interspecies spermatogonial transplantation: influence of recipient age on restoration of spermatogenesis by donor cells |
| Grantee: | Ricardo Jose Garcia Pereira |
| Support Opportunities: | Regular Research Grants |