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A Kinesin Is Present at Unique Sertoli/Spermatid Adherens Junctions in Rat and Mouse Testes¹

Department of Cellular and Physiological Sciences,⁴ Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3 Department of Biological Sciences,⁵ Faculty of Science, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6 Centre for Molecular Medicine and Therapeutics,⁶ University of British Columbia, Vancouver, British Columbia, Canada V5Z 4H4

ABSTRACT

During spermatogenesis, spermatids undergo a "down and up" translocation event in the seminiferous epithelium. This event has been proposed to result from the movement of ectoplasmic specializations, which are formed in Sertoli cells at sites of adhesion to spermatids, along adjacent microtubule tracts. To test the hypothesis that a kinesin is associated with ectoplasmic specializations, we generated antibodies to conserved kinesin sequences and detected kinesins on fixed frozen testis sections and fixed seminiferous epithelial fragments. The antibodies reacted with ectoplasmic specializations related to spermatids, in addition to reacting with other structures in the epithelium known to contain kinesins. At the electron microscopy level, the antibodies reacted with the cytoplasmic face of the endoplasmic reticulum component of ectoplasmic specializations. Based on mRNA transcript screens using mouse GeneChip arrays of testis and Sertoli cells, we identified KIF20 as a candidate kinesin at ectoplasmic specializations. Antibodies generated against a peptide sequence unique to this kinesin reacted at ectoplasmic specializations in testis sections and epithelial fragments, as well as with the endoplasmic reticulum component of ectoplasmic specializations when analyzed by electron microscopy. The antibody reacted on Western blots with full-length KIF20. On Western blots of testis lysates, the antibody reacted with a protein that is not present in other tissues and which migrates at a higher molecular weight than that predicted for KIF20. Our results demonstrate that a kinesin is associated with apical ectoplasmic specializations in Sertoli cells and that the motor may be an isoform of KIF20.

adherens junctions, ectoplasmic specializations, kinesin, Sertoli cells, spermatid translocation, testis

³These authors contributed equally to this work.

¹Supported by a Canadian Institutes of Health Research grant (MOP 62728) to A.W.V.

Correspondence: ²A. Wayne Vogl, Department of Cellular and Physiological Sciences, Division of Anatomy and Cell Biology, Faculty of Medicine, Life Sciences Centre, 2350 Health Science Mall, Vancouver, BC, Canada V6T 1Z3. FAX 604 822 2395; e-mail: vogl@interchange.ubc.ca

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