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Related Experiment Videos

Sperm plasma membrane integrity measurement: a combined method.

A Carreras1, J P Ramirez, C Mendoza

  • 1Departamento de Bioquímica y Biologia Molecular, Facultad de Ciencias, Universidad de Granada, Spain.

Andrologia
|November 1, 1992
PubMed
Summary

The hypo-osmotic swelling (HOS) test combined with eosin Y or propidium iodide staining effectively assesses sperm membrane integrity. High percentages of HOS-negative, stained sperm (types 2 and 4) indicate poor outcomes in in vitro fertilization (IVF).

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Area of Science:

  • Reproductive Biology
  • Sperm Physiology
  • In Vitro Fertilization

Background:

  • Sperm plasma membrane integrity is crucial for male fertility.
  • Assessing sperm viability and function aids in diagnosing infertility and optimizing assisted reproductive technologies.
  • The hypo-osmotic swelling (HOS) test is a functional assay for sperm membrane status.

Purpose of the Study:

  • To evaluate a combined hypo-osmotic swelling (HOS) test with eosin Y or propidium iodide staining for assessing sperm plasma membrane characteristics.
  • To correlate sperm membrane integrity, assessed by the HOS-dye tests, with lipid peroxidation levels.
  • To determine the prognostic value of the HOS-dye tests in predicting fertilization success in an in vitro fertilization (IVF) program.

Main Methods:

Related Experiment Videos

  • Sperm samples were fractionated using a swim-up technique to isolate motile spermatozoa.
  • The hypo-osmotic swelling (HOS) test was performed in conjunction with eosin Y (HOS-eosine) or propidium iodide (HOS-propidium) staining.
  • Lipid peroxidation levels were measured in different sperm fractions.
  • Spermatozoa were classified into four types based on HOS response and dye uptake.

    • Main Results:

    • The HOS-dye tests differentiated sperm based on membrane integrity, categorizing them into four types.
    • Fractionated motile sperm (Fraction II) exhibited higher percentages of intact membranes (Type 1) and lower lipid peroxidation compared to unselected sperm (Fraction I).
    • Non-motile sperm fractions (Fraction III) showed compromised membrane integrity (higher percentages of Types 2, 3, and 4) and elevated lipid peroxidation.
    • Preliminary IVF data indicated that a high incidence of Types 2 and 4 spermatozoa was associated with fertilization failure.

    Conclusions:

    • The combined HOS-dye test is a valuable method for assessing sperm plasma membrane integrity.
    • Sperm membrane integrity, as indicated by HOS-dye test results, correlates with lipid peroxidation levels.
    • The HOS-eosine test shows prognostic potential for predicting fertilization outcomes in IVF, with high rates of specific sperm types suggesting potential failure.