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Assessment of boar sperm intracellular Ca²⁺ level and motility characters by flow cytometry and CASA. Optimizing the protocol for Fluo-4 assessment of sperm intracellular Ca²⁺level by flow cytometry and investigation of relationships between storage time, intracellular Ca²⁺ and sperm motility characters

Khezri, Abdolrahman
Master thesis
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Khezri.pdf (4.098Mb)
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http://hdl.handle.net/11250/132359
Utgivelsesdato
2013
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  • Master i næringsrettet bioteknologi / Master of applied and commercial biotechnology M2NRBIOTEK [40]
Sammendrag
Porcine production industry demand effective artificial insemination for future challenges.

Variation in field fertility and litter size not only caused by sows or farm management but

also affected by semen and boar related parameters. Assessment of semen using objective

methods provides an effective tool for better and precise assessment of sperm quality during

the collection until consumption and leads to prediction of field fertility and genetic

selection.

In order to evaluate sperm intracellular Ca²⁺ level by flow cytometry, a Fluo-4 assay was

successfully adapted for boar spermatozoa by minimizing the handling effect and optimizing

the Fluo-4 incubation and concentration. Clear different Fluo-4 patterns were observed for

spermatozoa with low and high intracellular Ca²⁺ by fluorescence microscope and flow

cytometry.

In the present study, semen samples from Norwegian Landrace and Duroc were analyzed by

flow cytometry for assessment of sperm intracellular Ca²⁺ level and CASA for sperm

motility parameters on the day of collection and after 4 days liquid preservation at 18 °C.

Results showed that in both Duroc and Landrace, proportion of sperm cells with high

intracellular Ca²⁺ were increased during the storage time. The amount of motile sperm cells

was remained constant during the preservation in Landrace and decreased in Duroc.

Furthermore, high degree of hyperactivated sperm cells were detected in Duroc on the day of

collection and hyperactivited sperm population decreased after 4 days preservation while,

Landrace sperm motility pattern developed toward hyperactivation. In addition, for Duroc

semen, hyperactivation significantly affected positively by motility and negatively by

intracellular Ca²⁺ . While, Landrace semen hyperactivity level was affected just by

intracellular Ca²⁺ level. The present finding provides further evidence supporting the effect

of breed on sperm physiology and behavior. Moreover, this study has shown that objective

methods for assessment of semen quality could count as valid, rapid and precise methods in sperm evaluation.
Beskrivelse
Mastergradsoppgave i næringsrettet bioteknologi, Avdeling for lærerutdanning og naturvitenskap, Høgskolen i Hedmark, 2013. Master of applied biotechnology.

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