Functional changes and motility characteristics of Japanese Black bull spermatozoa separated by Percoll

K. Suzuki, M. Geshi, N. Yamauchi, T. Nagai

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Spermatozoa from two Japanese Black bulls (Bull-ATF and Bull-KTG) were separated by centrifugation at 700 × g for 15 min in modified TALP with or without 45-90% Percoll. Control washed spermatozoa and those collected from the bottom of 45 and 90% Percoll fractions were examined for viability and membrane integrity (using Hoechst bis-benzimide 33258 or propidium iodide and 6-carboxyfluorescein diacetate (PI-CFDA)), acrosomal status (using fluorescence isothiocyanate (FITC) conjugated Pisum Sativum agglutinin (PSA) and Peanut agglutinin (PNA), Naphthol Yellow S and Erythrosin B (NE) or triple staining (TS)), capacitation status (using chlortetracycline (CTC)), motility characteristics (using a computer-assisted sperm motion analysis system (CASA)) and for in vitro fertility. Percoll-separated spermatozoa showed greater viability and membrane integrity than controls, as determined by supravital staining. Differences were observed in the results regarding viability and acrosomal status of spermatozoa among sperm staining methods. Bull-ATF, which showed significantly greater in vitro fertility than Bull-KTG (P < 0.05), showed a significantly higher rate of CTC-B-pattern (capacitated) spermatozoa (P < 0.01) than Bull-KTG. The motility characteristics of control washed spermatozoa and those separated by 45-90% Percoll were analyzed by CASA. More motile and progressively motile spermatozoa were observed in the fraction at the bottom of the 90% Percoll solution than in the 45% Percoll fraction or in controls (P < 0.01). Moreover, the spermatozoa of Bull-KTG, which showed lower in vitro fertility than Bull-ATF, did not show significant differences in motility from those of Bull-ATF. These results provided basic information about Japanese Black bull spermatozoa, and suggested that spermatozoa with greater motility and viability can be obtained by Percoll separation than without separation. However, Percoll separation did not enhance their in vitro fertility.

Original languageEnglish
Pages (from-to)157-172
Number of pages16
JournalAnimal Reproduction Science
Volume77
Issue number3-4
DOIs
Publication statusPublished - Jul 15 2003

Fingerprint

Wagyu
Spermatozoa
bulls
spermatozoa
Fertility
viability
Chlortetracycline
chlortetracycline
agglutinins
Staining and Labeling
Percoll
Erythrosine
erythrosine
naphthols
Peanut Agglutinin
Membranes
Propidium
Agglutinins
Peas
isothiocyanates

All Science Journal Classification (ASJC) codes

  • Food Animals
  • Animal Science and Zoology
  • Endocrinology

Cite this

Functional changes and motility characteristics of Japanese Black bull spermatozoa separated by Percoll. / Suzuki, K.; Geshi, M.; Yamauchi, N.; Nagai, T.

In: Animal Reproduction Science, Vol. 77, No. 3-4, 15.07.2003, p. 157-172.

Research output: Contribution to journalArticle

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abstract = "Spermatozoa from two Japanese Black bulls (Bull-ATF and Bull-KTG) were separated by centrifugation at 700 × g for 15 min in modified TALP with or without 45-90{\%} Percoll. Control washed spermatozoa and those collected from the bottom of 45 and 90{\%} Percoll fractions were examined for viability and membrane integrity (using Hoechst bis-benzimide 33258 or propidium iodide and 6-carboxyfluorescein diacetate (PI-CFDA)), acrosomal status (using fluorescence isothiocyanate (FITC) conjugated Pisum Sativum agglutinin (PSA) and Peanut agglutinin (PNA), Naphthol Yellow S and Erythrosin B (NE) or triple staining (TS)), capacitation status (using chlortetracycline (CTC)), motility characteristics (using a computer-assisted sperm motion analysis system (CASA)) and for in vitro fertility. Percoll-separated spermatozoa showed greater viability and membrane integrity than controls, as determined by supravital staining. Differences were observed in the results regarding viability and acrosomal status of spermatozoa among sperm staining methods. Bull-ATF, which showed significantly greater in vitro fertility than Bull-KTG (P < 0.05), showed a significantly higher rate of CTC-B-pattern (capacitated) spermatozoa (P < 0.01) than Bull-KTG. The motility characteristics of control washed spermatozoa and those separated by 45-90{\%} Percoll were analyzed by CASA. More motile and progressively motile spermatozoa were observed in the fraction at the bottom of the 90{\%} Percoll solution than in the 45{\%} Percoll fraction or in controls (P < 0.01). Moreover, the spermatozoa of Bull-KTG, which showed lower in vitro fertility than Bull-ATF, did not show significant differences in motility from those of Bull-ATF. These results provided basic information about Japanese Black bull spermatozoa, and suggested that spermatozoa with greater motility and viability can be obtained by Percoll separation than without separation. However, Percoll separation did not enhance their in vitro fertility.",
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