ENZYMORPHOLOGY OF ESTROGEN 

 STIMULATED RAT VAGINA 



M. HAYASHI and W. H. FISHMAN 



Tufts University School of Medicine and the New England 

 Center Hospital, Boston, Massachusetts 



The /S-glucuronidase activity of the secondary sex tissues of the female 

 rodent is sensitive to the level of circulating estrogen. For purposes of 

 defining the exact locus of this enzyme-hormone relation, our previous i 

 experiments {Histochemical Society, April 9, 1960) demonstrated that ' 

 striking changes were évident as early as two hours following estrogen 

 injection by the subcutaneous route. At this time, jS-glucuronidase con- 

 centrated on the nuclei of cells of the vaginal stratum germinativum, 

 esterase increased in the cytoplasm of cells in the germinal and interme- 

 diate layers of the epithelium, and alkaline phosphatase enriched the 

 capillary endothelium. Altérations in acid phosphatase and DPNH- 

 -diaphorase took longer to appear. 



In the présent study, it has been possible to detect enzymorphologic 

 change with regard to vaginal j6-glucuronidase as early as 30 minutes 

 following intravenous injection of estrogen. Thus, five weeks after ovariec- 

 tomy, four— month old female Wistar rats received 2 to 10 y/100 g body 

 weight of estradiol injected into the fémoral vein. TTiey were sacrificed 

 at intervais rangin g from 5, 10, 30, to 480 minutes. The vagina was removed 

 and fixed immediately in formal-chloral fixative. Next morning 2 mm thick I 

 portions of the tissue were washed well with water (1 hr), embedded in 

 gelatin, and then returned to fresh fixative for another night. After washing 

 out the fixative (0*5 to 1*0 hr.), 7 micron thick sections were eut on the m 

 freezing microtome for enzymorphology, and cellular sites of activity were 

 demonstrated for /3-glucuronidase and «-naphthyl esterase. The first 

 remarkable enzyme change was the appearance of /S-glucuronidase in the 

 cytoplasm of the most superficiel layer of vaginal cells which now were 

 columnar in appearance. Moreover, the enzyme appeared to migrate towards 

 the lumenal surface. Photomicrographs will illustrate this event and ■ 

 others observed later with «-naphthyl esterase. Feulgen counterstained 

 préparations of thèse sections will also be presented and, where possible, 

 électron microphotographs. 



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