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Assessment of Sperm Function Parameters and DNA Fragmentation in Ejaculated Alpaca Sperm (Lama Pacos) by Flow Cytometry

Authors

  • C Cheuquemán,

    1. Scientific and Technological Bioresource Nucleus (BIOREN-Center of Reproductive Biotechnology), Universidad de La Frontera, Temuco, Chile
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  • O Merino,

    1. Scientific and Technological Bioresource Nucleus (BIOREN-Center of Reproductive Biotechnology), Universidad de La Frontera, Temuco, Chile
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  • L Giojalas,

    1. Centre for Cell and Molecular Biology, National University of Cordoba, Cordoba, Argentina
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  • A Von Baer,

    1. Llamas del Sur Farm, Jardín del Edén, Araucanía, Chile
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  • R Sánchez,

    1. Scientific and Technological Bioresource Nucleus (BIOREN-Center of Reproductive Biotechnology), Universidad de La Frontera, Temuco, Chile
    2. Department of Preclinical Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco, Chile
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  • J Risopatrón

    Corresponding author
    1. Department of Basic Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco, Chile
    • Scientific and Technological Bioresource Nucleus (BIOREN-Center of Reproductive Biotechnology), Universidad de La Frontera, Temuco, Chile
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Author's address (for correspondence): J Risopatrón, Centre of Reproductive Biotechnology, Avda. Francisco Salazar 01145, Universidad de La Frontera, Temuco-Chile. E-mail: jennie@ufro.cl

Contents

Flow cytometry has been shown to be an accurate and highly reproducible tool for the analysis of sperm function. The main objective of this study was to assess sperm function parameters in ejaculated alpaca sperm by flow cytometry. Semen samples were collected from six alpaca males and processed for flow cytometric analysis of sperm viability and plasma membrane integrity using SYBR-14⁄PI staining; acrosomal membrane integrity using FITC-conjugated Pisum Sativum Agglutinin⁄PI labelling; mitochondrial membrane potential (Δψm) by staining with JC-1 and DNA Fragmentation Index (DFI) by TUNEL. The results indicate that the mean value for sperm viability was 57 ± 8 %. Spermatozoa with intact acrosome membrane was 87.9 ± 5%, and viable sperm with intact acrosomal membrane was 46.8 ± 9%, high mitochondrial membrane potential (Δψm) was detected in 66.32 ± 9.51% of spermatozoa and mean DFI value was 0.91 ± 0.9%. The DFI was inversely correlated with high Δψm (p = 0.04; r = −0.41) and with plasma membrane integrity (p = 0.01; r = −0.47). To our knowledge, this is the first report of the assessment on the same sample of several parameters of sperm function in ejaculated alpaca sperm by flow cytometry.

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