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An in Vivo Study on Adjudin and Blood-Testis Barrier Dynamics

Ilona A. Kopera¹, Linlin Su¹, Barbara Biliska, C. Yan Cheng and Dolores D. Mruk

Population Council (I.A.K., L.S., C.Y.C., D.D.M.), Center for Biomedical Research, New York, New York 10065; and Department of Endocrinology and Tissue Culture (B.B.), Institute of Zoology, Jagiellonian University, 30-060 Krakow, Poland

Address all correspondence and requests for reprints to: Dolores D. Mruk, Ph.D., Population Council, Center for Biomedical Research, 1230 York Avenue, New York, New York 10065. E-mail: mruk{at}popcbr.rockefeller.edu.

Adjudin is known to specifically affect Sertoli-germ cell adhesion, resulting in germ cell loss from the seminiferous epithelium and transient infertility. The apical ectoplasmic specialization (ES) was shown to be the primary target of adjudin because adhesion was unaffected in organs that lack this structure. Herein we expand previous findings by treating rat pups with adjudin, and we aimed to address two questions. First, can adjudin perturb germ cell adhesion in the seminiferous epithelium of testes in which the apical ES is not yet present? Second, can adjudin affect assembly of the blood-testis barrier (BTB) at 15–18 d of age? Interesting changes were noted when aged-matched testes from control and adjudin-treated rats were examined, including a delay in the appearance of developing germ cells as well as a delay in the formation of the tubule lumen. Immunoblotting using antibodies against BTB-constituent proteins indicated that formation of the BTB was affected in rat pups gavaged with adjudin. These results were corroborated by immunofluorescence microscopy, which showed profound changes in the cellular distribution of tight junction and basal ES proteins. Moreover, the BTB was shown to be compromised in 30-d-old rats when its integrity was assessed by a functional in vivo assay. By 45 d of age, however, the seminiferous epithelium of treated rats was indistinguishable from that of control rats. Collectively these results demonstrate that adjudin targets the apical ES as well as the basal ES and tight junction, which in turn delays assembly of the BTB.