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Enzyme Activity and Immunohistochemical Localization of Dipeptidyl Peptidase 8 and 9 in Male Reproductive Tissues

Véronique Dubois, Chris Van Ginneken, Hilde De Cock, Anne-Marie Lambeir, Pieter Van der Veken, Koen Augustyns, Xin Chen, Simon Scharpé and Ingrid De Meester

Laboratory of Medical Biochemistry (VD,A-ML,SS,IDM), Anatomy/Embryology of Domestic Animals (CVG,HDC), and Laboratory of Medicinal Chemistry (PVDV,KA), University of Antwerp, Wilrijk, Belgium, and Division of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Zhu Nan, Taiwan, China (XC)

Correspondence to: Ingrid De Meester, Laboratory of Medical Biochemistry, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. E-mail: ingrid.demeester{at}ua.ac.be

The mRNA expression pattern of dipeptidyl peptidase (DPP) 8 and DPP9, two DPP4 homologs, was studied previously and showed a broad tissue distribution. In this study, protein expression and activity of DPP8 and DPP9 were investigated in male reproductive tissues of different mammals. Based on specific DPP activities and inhibition profiles, the proline-selective DPP activity in the bovine and rat testis could predominantly be attributed to DPP8/9 and not to DPP4. This is in contrast to the epididymis, where most of the activity was caused by DPP4. Bovine sperm preparations had very low or undetectable DPP8/9 activity. After characterization of polyclonal antibodies specific for DPP8 or DPP9, we could localize both enzymes in seminiferous tubules of the testis. A specific staining for DPP9 was found associated with spermatozoids embedded in the epithelium, just before their release into the lumen, and in spermatids. DPP8 was localized in spermatozoids in an earlier stage of maturation. These findings help to provide insight into the physiological role of DPP4-like enzymes in the male reproductive system. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. (J Histochem Cytochem 57:531–541, 2009)

Key Words: dipeptidyl peptidase • DPP IV • DPP4 • DPP2 • DPP8 • DPP9 • testis • epididymis • anti-DPP8 antibody • anti-DPP9 antibody

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