Document Type

Article

Publication Date

1-1-1994

Abstract

ABSTRACT: This study was to verify the usefulness of Nanovid microscopy techniques for evaluating induced modifications in bovine spermatozoal membranes. Frozen thawed bovine sperm were labeled with 20‐nm colloidal gold particles bound to concanavalin A (ConA) or heparin ligands. Sperm membrane changes were induced in vitro by capacitating and acrosome‐reacting procedures. Capacitatton was induced by incubation with 10 μg/ml of heparin for 4 hours at 37°C, 5% CO2, and high humidity. Membrane changes associated with the acrosome reaction were induced by addition of lysophosphatidyicholine (100 μg/ml) and incubation for 15 minutes at 37°C, 5% CO2, and high humidity. Gray intensity (black = 0; white = 255) of sperm (ONCELL) and background (OFFCELL) were evaluated with computer‐enhanced videomicroscopy with either differential interference contrast (DIC) or Nanovid optics. A high gold concentration on a membrane region produced blacker video pictures with Nanovid microscopy. Gray intensity of video pictures of a region with little or no gold would have a gray intensity equal to or greater than that of the background, that is, toward white. Weighted least squares methods were used to analyze ONCELL data using OFFCELL as a covariate. In experiment 1, ONCELL intensities of cells labeled with ConA‐gold complex were lower than those labeled with heparin‐gold at the same treatment level. In experiment 2, ONCELL intensity decreased as the concentration of heparin—gold increased from 0 to 0.041 μg/μl heparin. ONCELL intensity significantly de‐creased after sperm were treated with the highest heparin‐gold level and acrosome reacted. Differences in ONCELL intensity of acrosome‐reacted sperm were smaller in experiment 1 than in experiment 2. Nanovid microscopy techniques resulted in lower ONCELL intensities than did DIC microscopy in both experiments. 1994 American Society of Andrology

Publication Source (Journal or Book title)

Journal of Andrology

First Page

462

Last Page

467

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