Changes in plasma membrane and acrosome integrity of frozen-thawed bovine spermatozoa during a 4 h incubation as measured by multicolor flow cytometry

S. Nagy, Triin Hallap, Anders Johannisson, Heriberto Rodriguez-Martinez

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Cryopreservation of bull semen is sub-optimal, causing cell death of a majority of spermatozoa. Even the surviving cells are affected post-thaw, either structurally or functionally. The aim of this study was to investigate the sequence of events that take place when sperm plasma membrane and acrosome deteriorate during a 4h incubation period post-thaw, with special attention paid to the acrosome status of dying cells. Frozen-thawed semen of six AI dairy bulls was used. Three straws per batch were pooled and incubated at 37°C. Sub-samples were taken at 30min intervals and stained with SYBR 14, propidium iodide (PI) and phycoerythrin-conjugated peanut agglutinin (PE-PNA). Plasma membrane and acrosome integrity were measured by flow cytometry. The experiment was repeated three times. Immediately after thawing, only 3.45% of the dying cells showed acrosomal exocytosis. This number increased dramatically during incubation, reaching 67% after 4h. Within the intact cell population, the overall decrease in viability and acrosome integrity was kept at five percentage points. Flow cytometry and the triple fluorochrome combination presented a detailed picture of the time course in plasma membrane and acrosome deterioration of frozen-thawed bull semen. The results are expected to be useful for monitoring new cryopreservation protocols.

Original languageEnglish
Pages (from-to)225-235
Number of pages11
JournalAnimal Reproduction Science
Volume80
Issue number3-4
DOIs
Publication statusPublished - Feb 2004

Fingerprint

Acrosome
acrosome
flow cytometry
Spermatozoa
Flow Cytometry
plasma membrane
spermatozoa
Cell Membrane
cattle
semen
Cryopreservation
Semen
cryopreservation
bulls
Phycoerythrin
cells
dairy bulls
Semen Preservation
Peanut Agglutinin
exocytosis

Keywords

  • Acrosome
  • Bovine sperm
  • Cryopreservation
  • Flow cytometry
  • Plasma membrane

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Biotechnology
  • Genetics

Cite this

Changes in plasma membrane and acrosome integrity of frozen-thawed bovine spermatozoa during a 4 h incubation as measured by multicolor flow cytometry. / Nagy, S.; Hallap, Triin; Johannisson, Anders; Rodriguez-Martinez, Heriberto.

In: Animal Reproduction Science, Vol. 80, No. 3-4, 02.2004, p. 225-235.

Research output: Contribution to journalArticle

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