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Toxicologic Pathology, Vol. 33, No. 2, 300-304 (2005)
DOI: 10.1080/01926230590896145
© 2005 Society of Toxicologic Pathology

Articles

Morphology of the Fetal Rat Testis Preserved in Different Fixatives

Paul Howroyd1,2, Renee Hoyle-Thacker1, Otis Lyght1, Delorise Williams1 and Elena Kleymenova1

1 CIIT Centers for Health Research, Research Triangle Park, North Carolina 27709, USA
2 Experimental Pathology Laboratories, Inc., Research Triangle Park, North Carolina 27709, USA

Correspondence: Address correspondence to: Elena Kleymenova, CIIT Centers for Health Research, 6 Davis Drive, Research Triangle Park, North Carolina 27709, USA; e-mail:ekleymenova{at}ciit.org

Histopathological examination of the testes of exposed fetuses and neonates is important in assessing the developmental effects of environmental toxins, including sex hormone modulators. Modified Davidson’s fluid (mDF) has been suggested as a superior substitute for Bouin’s fluid for fixation of adult animal testes. We compared the morphology of fetal rat testes stained with hematoxylin and eosin (H&E) or immunochemically after fixation in 10% neutral buffered formalin (NBF), Bouin’s fluid, or mDF. Fixation in mDF resulted in more sharply defined nuclear detail and better preservation of cellular cytoplasm on H&E-stained sections of rat testes on gestation day 19. Use of Bouin’s fluid did not allow satisfactory detection of apoptotic cells by fluorescent terminal deoxynucleotide transferase-mediated deoxy-UTP nick labeling. Staining with the immunoperoxidase system and the conventional chromogen diaminobenzidine tetrahydrochloride to visualize 5-bromo-2-deoxyuridine-positive cells demonstrated that the number of positive nuclei and intensity of staining were similar with all 3 fixatives. Immunostaining for cytoskeletal protein vimentin was more intense and provided better details of the Sertoli cell cytoplasm with formalin fixation than with mDF. Our study demonstrates that fixation in mDF provided better morphologic detail in the fetal rat testis compared with 10% NBF and Bouin’s fluid and illustrates the importance of establishing the correct fixation conditions for each immunostaining protocol.

Key Words: Histology • fetus • seminiferous epithelium • nuclei • cellular cytoplasm • epifluorescence

Abbreviations: mDF, modified Davidson’s fluid • NBF, neutral buffered formalin • H&E, hematoxylin and eosin • gd, gestation day • TUNEL, terminal deoxynucleotide transferase-mediated deoxy-UTP nick labeling • DAB, diaminobenzidine tetrahydrochloride


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