Effect of Hoechst 33342 on stallion spermatozoa incubated in KMT or Tyrodes modified INRA96

Effect of Hoechst 33342 on stallion spermatozoa incubated in KMT or Tyrodes modified INRA96

The only known means of effectively separating populations of X and Y bearing sperms is the Beltsville sexing technology. The technology implies that each individual sperm is interrogated for DNA content, measuring the intensity of the fluorescence after staining the spermatozoa with Hoechst 33342....

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Veröffentlicht in:Animal reproduction science 2012-04, Vol.131 (3-4), p.165-171
Hauptverfasser: Balao da Silva, C, Macías-García, B, Morillo Rodriguez, A, Gallardo Bolaños, J.M, Tapia, J.A, Aparicio, I.M, Morrell, J.M, Rodriguez-Martínez, H, Ortega-Ferrusola, C, Peña, F.J
Format: Artikel
Sprache:eng
Schlagworte:
Animal and Dairy Science
Animals
Benzimidazoles - pharmacology
CASA
Cytoprotection - drug effects
DNA
flow cytometry
fluorescence
Fluorescent Dyes - pharmacology
Hoechst 33342
Horses - physiology
Husdjursvetenskap
Male
MEDICIN
MEDICINE
Sex Preselection - veterinary
Sex sorting
Sperm
sperm motility
Sperm Motility - drug effects
spermatozoa
Spermatozoa - drug effects
Staining and Labeling
Stallion
stallions
Time Factors
Veterinary Science
Veterinärmedicin
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Zusammenfassung:The only known means of effectively separating populations of X and Y bearing sperms is the Beltsville sexing technology. The technology implies that each individual sperm is interrogated for DNA content, measuring the intensity of the fluorescence after staining the spermatozoa with Hoechst 33342. Because there are no data regarding the effect of the staining on stallion sperm, ejaculates were incubated up to 90min in presence of 0, 4.5, 9, 22.5, 31.5, 45, 54, 67.5, 76.5 and 90μM of Hoechst 33342, in two media, KMT or INRA-Tyrodes. After 40 and 90min of incubation, motility (CASA) and membrane integrity (flow cytometry after YoPro-1/Eth staining) were evaluated. In KMT extender sperm motility significantly decreased after 45min of incubation when sperm were incubated in the presence of concentrations of Hoechst of 45μM or greater (P
ISSN:0378-4320
1873-2232
1873-2232
DOI:10.1016/j.anireprosci.2012.01.003