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Protocadherin 3 Acts at Sites Distinct from Classic Cadherins in Rat Testis and Sperm¹

Department of Pathology and Laboratory Medicine, Brown University, Providence, Rhode Island 02912

The testis expresses a variety of cadherin superfamily members including classic cadherins and protocadherins. This report describes the first localization of a protocadherin protein in testis and sperm. After cloning rat cDNAs for protocadherin 3 and 4, isoform-specific polyclonal antibodies were generated against protocadherin 3. Western blotting of rat testis showed that protocadherin 3 was solubilized completely by Triton X-100, in contrast to the adhesion junction components N-cadherin, ß-catenin, and p120 catenin. Corroborating this data, protocadherin 3 was immunolocalized to the spermatid acrosomal area, intercellular bridge, and flagellum, but not classic cadherin-based adhesion junctions. Acrosome-associated protocadherin 3 was first detected at step 8 of spermiogenesis, and this association remained on cauda epididymal sperm. Acrosome immunostaining was reduced, but present, in acrosome-reacted sperm. Spermatid intercellular bridges became positive for protocadherin 3 coincident with the appearance of plectin, occurring at spermiogenic steps 8 to 9, and elongate spermatid bridges remained positive throughout spermatogenesis. The developing flagellum was uniformly immunostained for protocadherin 3 up to approximately spermiogenic step 17. Subsequently, flagellar immunostaining was confined to the principal piece, and this pattern continued in cauda epididymal sperm. These data show that protocadherin 3 performs functions unique from classic cadherins in spermatogenesis and suggest a role for protocadherin 3 in organizing germ cell-specific structures including the intercellular bridge, flagellum, and acrosome.

¹ Supported by Public Health Service grant NIEHS RO1 ES11632-01 and grant RR16457 from the BRIN program of the National Center for Research Resources awarded to K.J.J.

² Correspondence: Kamin J. Johnson, CIIT Centers for Health Research, 6 Davis Drive, Research Triangle Park, NC 27709. FAX: 919 558 1300; kjohnson{at}ciit.org

³ Current address: CIIT Centers for Health Research, 6 Davis Drive, Research Triangle Park, NC 27709

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