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Cystatin C Is Highly Expressed in the Human Male Reproductive System

MAGNUS ABRAHAMSON

HANNA WALLIN

JOHAN MALM

ÅKE LUNDWALL

VIRGIL GADALEANU

From the^* Department of Urology, Malmö University Hospital, Lund University, Sweden; the Department of Clinical Chemistry, Lund University Hospital, Lund University, Sweden; and the Department of Laboratory Medicine, Divisions of Clinical Chemistry and Pathology, Malmö University Hospital, Lund University, Sweden.

Correspondence to: Dr Thomas Jiborn, Department of Urology, Malmö University Hospital, 205 02 Malmö, Sweden (e-mail: thomas.jiborn{at}skane.se).

Cystatin C displays the strongest inhibitory activity of all cystatins toward lysosomal cysteine proteases in general and has a widespread distribution in human tissues and body fluids, including seminal plasma. The aim of this study was to investigate the distribution of cystatin C in the male reproductive system. Immunohistochemistry revealed a widespread distribution of cystatin C in normal tissues from the testis, epididymis, vas deferens, seminal vesicle, and prostate gland. Immunoreactive cystatin C was localized in basal and secretory epithelial cells, but also in neuroendocrine cells in the prostate, identified by immunostaining for chromogranin A. On adjacent tissue sections, we demonstrated local production of cystatin C utilizing nonradioactive in situ hybridization with a 201-base-long digoxigenin-labeled antisense RNA probe specific for the cystatin C transcript. Staining patterns obtained by immunohistochemistry and in situ hybridization correlated well. Enzyme-linked immunosorbent assay for quantitative analysis of cystatin C demonstrated that cystatin C was present at high concentrations in tissue homogenates from all locations investigated, compared to liver, muscle, spleen, and other general tissues. Western blotting of tissue homogenates revealed a predominant 15-kd cystatin C immunoreactive component in accordance with previous findings in other organs. Quantitative real-time polymerase chain reaction analysis to determine messenger RNA levels in whole tissue extracts showed that the cystatin C gene is highly expressed in the seminal vesicles and the prostate gland, indicating that the major amount of cystatin C in the male reproductive organs and seminal plasma is produced by cells in these 2 tissues. It is concluded that cystatin C is highly expressed and widely distributed throughout the male genital tract, suggesting that cystatin C is an important regulator for normal and pathological proteolysis in the male reproductive system.

     Key words: Cysteine protease inhibitor, immunohistochemistry, in situ hybridization, quantitative real-time (QRT) PCR, seminal fluid

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