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CD26 and Adenosine Deaminase Interaction: Its Role in the Fusion Between Horse Membrane Vesicles and Spermatozoa¹

^a Dipartimento di Biologia Cellulare e Molecolare, Università di Perugia, 06123 Perugia, Italia ^b Centrum for Laboratorie Medicine, Akademiska sjukhuset, Uppsala, Sweden ^c Departament de Bioquimica i Biologia Molecular, Universitat de Barcelona, Barcelona, España

Membrane vesicles of horse seminal plasma present at their surface a highly specific serine-type protease, dipeptidyl peptidase IV/CD26, a surface antigen known to characterize human prostasomes.

Horse sperm cells expressed at their surface A₁ adenosine receptors (A₁AR) and ecto-adenosine deaminase (ecto-ADA), both detected by immunoblot analysis, whereas CD26 was visualized at the equatorial segment by immunofluorescence microscopy. In addition to CD26, horse membrane vesicles showed ecto-ADA. The fusion process between horse sperm cells and vesicles was evidenced by confocal microscopy, which showed the localization of CD26 at the postacrosomal region and at the midpiece of the spermatozoa after incubation with vesicles. Moreover, a similar localization of CD26 and ecto-ADA on the spermatozoa was evidenced after fusion.

Our results suggest that the interaction CD26/ecto-ADA might be responsible for fusion. Since A₁ARs are said to be second receptors for ecto-ADA to form ecto-ADA/A₁AR complexes, and since horse spermatozoa have A₁ARs at their surface, the interaction CD26/ecto-ADA/A₁AR during the fusion process cannot be ruled out.

¹ This research was supported by National Research Council of Italy, MURST 40%, Spanish CICYT (PB94/0941 and PB97/0984), and Spanish CICYT's I+D program Salud y Farmacia (SAF97/0066).

² Correspondence: Alba Minelli, Dipartimento di Biologia Cellulare e Molecolare, Sezione Enzimologia, Università di Perugia, Via del Giochetto, 06123 Perugia, Italia. FAX: 39 75 5853442; albami{at}tin.it

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