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Home / Equine Research Bank / J Cell Sci / Glyceraldehyde 3-phosphate dehydrogenase is bound to the fib...
Journal of cell science1997; 110 ( Pt 15); 1821-1829; doi: 10.1242/jcs.110.15.1821

Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa.

Abstract: Evidence is provided that the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase is covalently linked to the fibrous sheath. The fibrous sheath is a typical structure of mammalian spermatozoa surrounding the axoneme in the principal piece of the flagellum. More than 90% of boar sperm glyceraldehyde 3-phosphate dehydrogenase activity is sedimented after cell disintegration by centrifugation. Detergents, different salt concentrations or short term incubation with chymotrypsin do not solubilize the enzyme, whereas digestion with trypsin or elastase does. Short term incubation with trypsin (15 minutes) even resulted in an activation of glyceraldehyde 3-phosphate dehydrogenase. Purification on phenyl-Sepharose yielded a homogeneous glyceraldehyde 3-phosphate dehydrogenase as judged from gel electrophoresis SDS-PAGE and native gradient PAGE. The molecular masses are 41.5 and 238 kDa, respectively, suggesting native glyceraldehyde 3-phosphate dehydrogenase to be a hexamer. Rabbit polyclonal antibodies raised to purified glyceraldehyde 3-phosphate dehydrogenase show a high specificity for mammalian spermatozoal glyceraldehyde 3-phosphate dehydrogenase, while other proteins of boar spermatozoa or the muscle glyceraldehyde 3-phosphate dehydrogenase are not labelled. Immunogold staining performed in a post-embedding procedure reveals the localization of glyceraldehyde 3-phosphate dehydrogenase along the fibrous sheath in spermatozoa of boar, bull, rat, stallion and man. Other structures such as the cell membrane, dense fibres, the axoneme or the mitochondria are free of label. During the process of sperm maturation, most of the cytoplasm of the sperm midpiece is removed as droplets during the passage through the epididymis. The labelling of this cytoplasm, in immature boar spermatozoa and in the droplets, indicates that glyceraldehyde 3-phosphate dehydrogenase is completely removed from the midpiece during sperm maturation in the epididymis. The inverse compartmentation of the glycolytic enzyme and mitochondria in the mammalian sperm flagella suggests that ATP-production in the principal piece mainly occurs by glycolysis and in the midpiece by respiration.
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Publication Date: 1997-08-01 PubMed ID: 9264469DOI: 10.1242/jcs.110.15.1821Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates the binding of the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase to the fibrous sheath of mammalian spermatozoa, suggesting a key role in energy production during sperm movement.

Research Methods and Findings

  • The researchers started by observing that after disintegration of boar sperm cells by centrifugation, over 90% of the glyceraldehyde 3-phosphate dehydrogenase activity was found in the sediment, indicating that this enzyme is predominantly located in the solid parts of the cell.
  • Attempts to release the enzyme from these parts using detergents, different salt concentrations, or chymotrypsin didn’t succeed significantly. However, treatment with trypsin or elastase was effective, which suggests a very firm binding. Trypsin also activated the enzyme, signaling the possibility of a physiological interaction.
  • Gel electrophoresis methods were used for purification and identification, showing that the enzyme is a hexamer with molecular masses of 41.5 and 238 kDa.
  • Rabbit polyclonal antibodies specifically recognized this exact spermatozoal form of the enzyme, confirming its unique identity.
  • Immunogold staining further confirmed the location of the enzyme along the fibrous sheath in the sperm of different mammals, while it was absent from other sperm and cellular structures.

Implications and Conclusions

  • The researchers noted that the enzyme also appeared in the immature sperm and was then removed during maturation, suggesting a regulatory role of the epididymis in producing mature sperm with optimized structure and function.
  • The specific location of the glyceraldehyde 3-phosphate dehydrogenase within the sperm structure, and the inverse relationship to the mitochondria, suggest it plays a central role in ATP, the primary energy source for cell function, generation within the principal piece of the sperm via glycolysis.
  • The study provided key insights into the importance of the exact position of key enzymes, such as the glyceraldehyde 3-phosphate dehydrogenase, in the functional differentiation of regions within the sperm, and how this may contribute to their motility and fertilizing capabilities.

Cite This Article

APA
Westhoff D, Kamp G. (1997). Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa. J Cell Sci, 110 ( Pt 15), 1821-1829. https://doi.org/10.1242/jcs.110.15.1821

Publication

Journal of cell science
ISSN: 0021-9533
NlmUniqueID: 0052457
Country: England
Language: English
Volume: 110 ( Pt 15)
Pages: 1821-1829

Researcher Affiliations

Westhoff, D
  • Institut für Zoophysiologie, Westfälische Wilhelms-Universität Münster, Germany.
Kamp, G

    MeSH Terms

    • Animals
    • Antibody Specificity
    • Cattle
    • Cytoplasm / enzymology
    • Electrophoresis, Polyacrylamide Gel
    • Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry
    • Glyceraldehyde-3-Phosphate Dehydrogenases / immunology
    • Glyceraldehyde-3-Phosphate Dehydrogenases / isolation & purification
    • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
    • Horses
    • Humans
    • Immunohistochemistry
    • Male
    • Mitochondria / enzymology
    • Molecular Weight
    • Rabbits
    • Rats
    • Solubility
    • Sperm Maturation
    • Sperm Tail / enzymology
    • Sperm Tail / ultrastructure
    • Swine

    Citations

    This article has been cited 35 times.
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