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Reproductive Cytotoxicity Is Predicted by Magnetic Resonance Microscopy and Confirmed by Ubiquitin–Proteasome Immunohistochemistry in a Theophylline-Induced Model of Rat Testicular and Epididymal Toxicity

Published online by Cambridge University Press:  07 July 2005

M.W. Tengowski ,
P. Sutovsky ,
L.W. Hedlund ,
D.J. Guyot ,
J.E. Burkhardt ,
W.E. Thompson ,
M. Sutovsky  and
G.A. Johnson
M.W. Tengowski
Affiliation:
Safety Sciences Groton, Pfizer Global Research & Development, Groton, CT 06340, USA
P. Sutovsky
Affiliation:
Departments of Animal Science and Obstetrics & Gynecology, University of Missouri-Columbia, Columbia, MO 65211, USA
L.W. Hedlund
Affiliation:
Center for In Vivo Microscopy, Duke University, Durham, NC 27710, USA
D.J. Guyot
Affiliation:
Safety Sciences Groton, Pfizer Global Research & Development, Groton, CT 06340, USA
J.E. Burkhardt
Affiliation:
Safety Sciences Groton, Pfizer Global Research & Development, Groton, CT 06340, USA
W.E. Thompson
Affiliation:
Cooperative Reproductive Science Research Center, Morehouse School of Medicine, Atlanta, GA 30310, USA
M. Sutovsky
Affiliation:
Departments of Animal Science and Obstetrics & Gynecology, University of Missouri-Columbia, Columbia, MO 65211, USA
G.A. Johnson
Affiliation:
Center for In Vivo Microscopy, Duke University, Durham, NC 27710, USA
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Abstract

This study investigated the testicular changes in the rat induced by the nonspecific phosphodiesterase inhibitor, theophylline using magnetic resonance microscopy (MRM) and ubiquitin immunostaining techniques. In vivo T1- and T2-weighted images were acquired at 2 T under anesthesia. Increased signal observed in the theophylline-treated rats suggests that leakage of MRM contrast was occurring. In vivo MRM results indicate that day 16 testis displayed an increased T1-weighted water signal in the area of the seminiferous tubule that decreased by day 32. These findings were validated by histopathology, suggesting that in vivo MRM has the sensitivity to predict changes in testis and epididymal tissues. The participation of the ubiquitin system was investigated, using probes for various markers of the ubiquitin-proteasome pathway. MRM can be used to detect subtle changes in the vascular perfusion of organ systems, and the up-regulation/mobilization of ubiquitin-proteasome pathway may be one of the mechanisms used in theophylline-treated epididymis to remove damaged cells before storage in the cauda epididymis. The combined use of in vivo MRM and subsequent tissue or seminal analysis for the presence of ubiquitin in longitudinal studies may become an important biomarker for assessing testis toxicities drug studies.

Keywords

magnetic resonance imagingmicroscopyreproductiontoxicologytheophyllinerattestisepididymisubiquitinproteasomeArylsulfatase A
Type
BIOLOGICAL APPLICATIONS
Information
Microscopy and Microanalysis , Volume 11 , Issue 4 , August 2005 , pp. 300 - 312
Copyright
© 2005 Microscopy Society of America

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