Immunostaining of human spermatozoa with tubulin domain-specific monoclonal antibodies. Recognition of a unique beta-tubulin epitope in the sperm head.
Four monoclonal antibodies that discriminate between structural domains of alpha-(TU-01, TU-04) or beta-(TU-06, TU-12) tubulin and a polyclonal anti-tubulin antibody were used for immunostaining of human spermatozoa using immunofluorescence microscopy. Specificity of antibodies was confirmed by immunoblotting experiments. Antibodies TU-01 and TU-06 uniformly stained the whole tail and the neck, whereas antibodies TU-04, TU-12 showed differential distribution of corresponding epitopes in the stable arrays of flagellar microtubules. Of the monoclonal antibodies used, only TU-12 against the antigenic determinant on C-terminal domain of beta-tubulin showed strong reactivity with the equatorial segment of the head. The results document a differential exposure of tubulin epitopes at the single-cell level and suggest the existence of distinct tubulin populations in various structural compartments of the human spermatozoon.
Dráber
P
P
Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Prague.
Dráberová
E
E
Viklický
V
V
D000911:Antibodies, Monoclonal / Q000276:immunology; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D008870:Microtubules / Q000737:chemistry; D013077:Sperm Head / Q000737:chemistry*; D013082:Sperm Tail / Q000737:chemistry; D013094:Spermatozoa / Q000737:chemistry*; D014404:Tubulin / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D014404:Tubulin