Scientists from the Lithuanian-Latinvian Veterinary Research Institute and the University of Helsinki have developed a simple fluorescence analysis method to evaluate boar semen used for artificial insemination.
A number of previous studies have shown that there is a correlation between sperm cell membrane integrity and fertilization rates. In this study published in the Acta Veterinaria Scandinavica, Sutkeviciene of the Lithuanian Veterinary Research Institute and other authors described a simple fluorescence analysis method. They used fluorescent DNA dyes to stain the mirror cells of artificially created cell membranes, and observed the fluorescence intensity.
method
The fluorescence intensity of the nuclei of the spermatozoa with disrupted cell membranes was measured using dyes Hoechst 33258 (H258) and propidium iodide (PI) that did not penetrate cell membranes. The fluorescence of intact sperm nuclei was measured using the dye Hoechst 33342 (H342) that penetrates the cell membrane. strength.
The sperm concentration in the artificial insemination single dose ranged from 31.2 106 to 50 106/ml, with an average of 3550 106/ml.
Three consecutive semen were collected for each boar, with a median interval of one week. The rate of unresponsiveness (NR%) and the number of litters born in the passed sows (the total number of litters) within 60 days after the first insemination were used as indicators of the conception rate.
result
The fluorescence intensity of the semen treated with H258 and H342 staining was significantly correlated with the litter size of the litter of the sows. The correlation of the fluorescence intensity of the PI staining was not significant, and H258 was r=-0.68 (P).
in conclusion
Membrane damage sperm H258 staining and membrane homozygous H342 stained fluorescence readings are associated with a decrease in litter size after artificial insemination.
This finding shows that the fluorescent properties of sperm nuclei can be used as an assessment of the fertility potential of artificial insemination semen, said Sutkeviciene and co-authors.
A number of previous studies have shown that there is a correlation between sperm cell membrane integrity and fertilization rates. In this study published in the Acta Veterinaria Scandinavica, Sutkeviciene of the Lithuanian Veterinary Research Institute and other authors described a simple fluorescence analysis method. They used fluorescent DNA dyes to stain the mirror cells of artificially created cell membranes, and observed the fluorescence intensity.
method
The fluorescence intensity of the nuclei of the spermatozoa with disrupted cell membranes was measured using dyes Hoechst 33258 (H258) and propidium iodide (PI) that did not penetrate cell membranes. The fluorescence of intact sperm nuclei was measured using the dye Hoechst 33342 (H342) that penetrates the cell membrane. strength.
The sperm concentration in the artificial insemination single dose ranged from 31.2 106 to 50 106/ml, with an average of 3550 106/ml.
Three consecutive semen were collected for each boar, with a median interval of one week. The rate of unresponsiveness (NR%) and the number of litters born in the passed sows (the total number of litters) within 60 days after the first insemination were used as indicators of the conception rate.
result
The fluorescence intensity of the semen treated with H258 and H342 staining was significantly correlated with the litter size of the litter of the sows. The correlation of the fluorescence intensity of the PI staining was not significant, and H258 was r=-0.68 (P).
in conclusion
Membrane damage sperm H258 staining and membrane homozygous H342 stained fluorescence readings are associated with a decrease in litter size after artificial insemination.
This finding shows that the fluorescent properties of sperm nuclei can be used as an assessment of the fertility potential of artificial insemination semen, said Sutkeviciene and co-authors.
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