Upsala J Med Sci 90: 139-145, 1985 Biochemical and Morphologic Studies of the Prostate Gland in Men Subjected to Radical Cystectomy Bernd Stegmayr,’ Christer Busch,’ b;ke Fritjofsson3 and Gunnar Ronquist4 ‘Department of Internal Medicine, Central Hospital, Eskilstuna and the ’Departments of Pathoiogy, 3Urology, and 4Clinical Chemistry, University Hospital, Uppsala, Sweden ABSTRACT Whole human p r o s t a t e g l a n d s o b t a i n e d from p a t i e n t s u n d e r g o i n g r a d i c a l c y s t - ectomy were d i s s e c t e d i n t o t h r e e p a i r e d l o b e s and t h e v a r i o u s p a r t s o f t h e g l a n d were s u b j e c t e d t o b i o c h e m i c a l and m o r p h o l o g i c e x a m i n a t i o n . No d i s t i n c t d i f f e r - ences were f o u n d between t h e p r o s t a t e l o b e s i n r e g a r d to c o n t e n t o f d i v a l e n t c a t i o n s , a c i d phosphatase and ATPase. These f i n d i n g s were c o n c o r d a n t w i t h ob- s e r v a t i o n s a t l i g h t microscopy. Hence, d e s p i t e d i s c e r n i b l e change i n c e l l u l a r appearance, no d i s t i n c t b o r d e r was o b s e r v e d between l o b e s i n d i c a t i n g s e p a r a t e “compar tmen t s “ INTRODUCTION A c i d phosphatase, Zn2+ and Mg2+ a r e w e l l known s e c r e t o r y p r o d u c t s i n human s e m i n a l plasma (4,5,6,9 ).An Mg2+ and Ca2+-dependent ATPase system was f o u n d t o be a n o t h e r i m p o r t a n t s e m i n a l component ( 1 0 ) . S t u d i e s o f s p l i t e j a c u l a t e r e v e a l e d a l l t h e s e p a r a m e t e r s t o be p a r t o f t h e human p r o s t a t i c s e c r e t i o n (1,lO). I n f u r - t h e r a n a l y s e s t h i s ATPase a c t i v i t y was f o u n d t o be a s s o c i a t e d w i t h t h e p r e s e n c e o f o r g a n e l l e s i n t h e s e m i n a l plasma ( 1 0 ) . The aim o f t h i s s t u d y was t o c l a r i f y more t h o r o u g h l y t h e e x t e n t t o w h i c h t h e s e o r g a n e l l e s , l a t e r d e n o t e d prostasomes ( 2 , 1 2 ) , and a l s o t h e o t h e r sub- s t a n c e s , a r e p r e s e n t i n and e x c r e t e d f r o m t h e human p r o s t a t i c g l a n d , and wheth- er l o c a l d i f f e r e n c e s w i t h r e g a r d t o c o n t e n t o f t h e s e s u b s t a n c e s e x i s t i n d i f f e r - e n t p a r t s o f t h e p r o s t a t e . MATERIAL AND METHODS P r o s t a t i c t i s s u e was o b t a i n e d f r o m seven men aged 5 9 - 68 y e a r s who were un- d e r g o i n g r a d i c a l c y s t e c t o m y , i n c l u d i n g t o t a l p r o s t a t e c t o m y , because o f b l a d d e r c a n c e r . None o f t h e men had had symptoms or s i g n s o f p r o s t a t i c d i s e a s e b e f o r e 10-858572 139 the operation. All had received preoperative irradiation to a dosage of 40 Gy over 3 weeks, and cystectomy and urinary diversion were performed 3 weeks after termination of radiotherapy. Immediately after the operation the prostatic gland was dissected free from the bladder, the seminal vesicles and adhering tissues. The prostate was then further dissected into three paired lobes as described by Tissell & Salander in 1975 (13). In three of the seven cases the prostatic tissue was immediately prefixed in isotonic glutaraldehyde (3 % ) buffer solut- ion (280 mosmol/l), kept at 4OC until fixation for electron microscopy with osmium tetroxide,and embedded in epon according to Ronquist et al. (10). In the other four cases all the material was immediately stored in a moist chamber at O0C and kept in a solution of ice and water to maintain temperature stability. After 4 to 7 hours representative specimens of prostatic tissue were carefully cut from this material and prefixed in glutaraldehyde for electron microscopy as outlined above. From the remaining prostatic cut sections,secretory fluid was gently squeezed from the ductules and collected in small plastic tubes. The fluid was diluted 1:3 with 0.15 M NaCl solution and centrifuged at 2 000 x g for 10 minutes. The residual pellet, containing cells and cell debris, was dis- carded and the supernatant was used for biochemical analyses. Magnesium, calcium and zinc were determined in an atomic absorption emission spectrophotometer. Acid phosphatase was measured colorimetrically according to a tartrate-inhibition method (7). Mg2+- and Ca2+-dependent ATPase activity was measured according to Ronquist et al. (10). The protein concentration was meas- ured with the method of Lowry et al. (8). To avoid influence of possible ex- ternal fluid on concentration, the parameters were related to the protein con- centration of the fluid. In three patients the prostatic dissection disclosed well-defined periureth- r a l hyperplastic nodules, the largest with a diameter of 20 mm. The nodules were removed from the prostatic tissue without rupture of the surrounding cap- sule. The solitary nodules likewise were sectioned for electron microscopy. Moreover, secretion was obtained by gentle squeezing for the aforementioned biochemical investigations. No contamination with prostatic tissue material was possible, as the nodules were separately prepared. RESULTS Biochemical investigations The values for acid phosphatase, calcium, magnesium and zinc were as expect- ed for prostatic fluid, although the analyzed specimens were obtained from squeezing of the dissected glandular material (Table 1). In all cases the Mg - 2+ 140 2 and Ca - d e p e n d e n t ATPase a c t i v i t y c o u l d r e a d i l y b e r e c o r d e d and was p r e s e n t a t a l e v e l h i g h e r t h a n t h a t e x p e c t e d f o r mixed s e m i n a l plasma ( c o n t a i n i n g s e c r e t i o n a l s o from s e m i n a l v e s i c l e s , t e s t e s and e p i d i d y m e s ) . The h y p e r p l a s t i c n o d u l e s d i s p l a y e d e s s e n t i a l l y t h e same c o n t e n t o f t h e s e s u b s t a n c e s a s t h e p r o s t a t i c t i s s u e . T a b l e 1. o f f l u i d from t h e p r o s t a t e g l a n d i n 4 men and i n adenornatous t i s s u e from 3 o f t h e same g l a n d s ATPase, a c i d p h o s p h a t a s e (AP), c a l c i u m ( C a ) , magnesium (Mg) and z i n c Ca Mg Zn ATPase -1 AP (nmol x min ( m k a t x g x g p r o t e i n ) p r o t e i n - 1 ) -1 ( A m 0 1 x g p r o t e i n - ’ ) P r o s t a t i c a l a n d mean 54.5 235 1 7 1 44.4 21.0 SDn-l 41.3 197 111 1 7 . 1 1 2 . 0 r a n g e 8 - 2 - 1 7 0 59.0-684 72.2-396 24.8-68.0 7.9-47.0 H y p e r p l a s t i c n o d u l e s m a n 50.1 1 2 9 . 3 1 3 5 54.2 21.6 Son-’ 1 6 . 5 5 3 . 5 48.7 29.6 5 . 3 r a n g e 38-6-69. o 9 1 .O-190 98-6-190 34.8-88.3 16.3-26.9 The c o n c e n t r a t i o n o f t h e s e s u b s t a n c e s v a r i e d t o some e x t e n t w i t h i n d i f f e r e n t p r o s t a t i c l o b e s , b u t w i t h o u t s t a t i s t i c a l l y s i g n i f i c a n t d i f f e r e n c e s ( T a b l e 2 ) . Nor was any mutual c o r r e l a t i o n found between t h e p a r a m e t e r s w i t h d i f f e r e n t l o c - a t i o n s i n t h e g l a n d . T a b l e 2. a s i n T a b l e 1. Means ( S O n - 1 ) ] V a l u e s f o r p r o s t a t i c f l u i d a c c o r d i n g t o l o b e o f o r i g i n [ a b b r e v i a t i o n s ATPase AP Ca Mg Zn Lobes from 4 g l a n d s D o r s a l L a t e r a l 4 0 . 9 258.9 1 7 8 . 0 4 4 . 1 23.2 ( 2 4 . 3 ) ( 2 1 2 . 9 ) ( 1 0 1 . 8 ) (20.8) ( 1 2 . 6 ) 75.7 298.0 144.0 4 0 . 1 21.9 ( 6 4 . 1 ) ( 2 6 9 . 0 ) ( 1 1 4 . 0 ) (11.1) (17.5) Medial 42.0 149.0 192.0 49.1 1 7 . 8 ( 1 5 . 1 ) ( 9 5 . 2 ) ( 1 4 1 . 0 ) ( 2 1 . 4 ) ( 6 . 2 ) 141 F r u c t o s e was n o t d e t e c t e d i n any o f t h e s e f l u i d sl g l a n d . Jecimens f r o m t h e p r o s t a t e F i g . 1 - A. S e c t i o n showing t h e d o r s a l l o b e p a t - t e r n . A c i n i a r r a n g e d i n a p a r a l l e l p a t t e r n w i t h s i m p l e p a p i l l a r y p r o j e c t i o n s ( o r i g i n a l m a g n i f i c a t i o n x 4 1 ) . B. L a t e r a l l o b e p a t - t e r n w i t h i r r e g u l a r a c i n i and c o a r s e stroma. N o t e t h e mild i n f l a m m a t o r y changes and s l o u g h i n g o f t h e g l a n d u l a r e p i t h e l i u m , p r o b a b l y due t o t h e p r e o p e r a t i v e i r r a d i - a t i o n ( x 4 1 ) . - C . M e d i a l l o b e p a t - t e r n w i t h p r o m i n e n t p a p i l l a r y f o r m a t i o n s ( x 41). - 142 Liaht microscow The morphologic pattern within each isolated lobe was generally variable (Fig. l), 1.e. did not show purely medial, lateral o r dorsal lobe chanacterist- ics as described by earlier authors (11). In two cases, however, pure patterns were observed, one with a typical dorsal lobe pattern and another with a typical lateral one. Transmission electron microscopy Fig. 2 shows the intracellular location of prostasomes within other, bigger organelles, the storage vesicles. Very few, if any, prostasomes are free in the cytoplasm (2). The size and general appearance of the intracellular prostasomes are the same as in prostasomes isolated from prostatic fluid and seminal plasma (2). DISCUSSION The patients from whom the tissue material was obtained had been irradiated preoperatively because of bladder cancer. As the radiotherapy was directed not only towards the bladder, but also to the pelvic lymph nodes, shedding irradi- ation could have involved the prostatic tissue used in our investigations. The -registered biochemical values thus may have been lower than in nonirradiated men, but this did not seem to be the case. Ultrastructural, atrophic changes may also be expected after radiotherapy. In the present study there were no distinct differences between the prostate lobes in regard to their content of divalent cations, acid phosphatase and ATP- ase. Convincing evidence has accumulated that this latter enzyme system is in- timately linked to the enveloping membrane of organelles (prostasomes) occurr- ing free in prostatic fluid and seminal plasma (10) as well as in prostatic tissue (secretory cells and acinar ducts, cf reference 2 and Fig. 2). The ATPase activity may function as a quantitative measure of the presence of prostasomes. These findings were concordant with the observations at light microscopy. Hence, although a change in cellular appearance was discernible, no distinct boundaries were observed between lobes that could indicate "com- partments". Our study also indicated similarities between hyperplastic and normal prost- atic tissue, although the hyperplastic tissue did not macroscopically present ducts connecting with other prostatic ducts. Earlier authors (14) showed that the zinc content/g tissue was at least equal in hyperplastic and normal prost- atic tissue. In o u r cases, moreover, the hyperplastic nodules were enveloped in a fibrous capsule. This investigation supports the view that hyperplastic tissue may develop from obstructed parts o f the true prostatic gland, because of the demonstrated similarities between the two tissue t y p e s . 143 144 Fig. 2 - A. Storage vesicles ( S ) in epithelial prostatic cells. The storage vesicles containing prostasomes are encased in a trilaminar membrane (x 2 7 000) - B. Higher magnification of membraneenveloped prostasomes (p) in storage vesicles ( x 6 7 0 0 0 ) REFERENCES 1. 2. 3 . 4. 5. 6. 7. 8. 9. 10. 11. 1 2 . 1 3 . 1 4 . Brody, I., Ronquist, G., Gottfries, A. & Stegmayr, B.: Abnormal deficiency of both Mg- and Ca-dependent adenosine triphosphatase and secretory granules and vesicles in human seminal plasma. Scand J Urol Nephrol 1 5 : 8 5 - 9 0 , 1 9 8 1 . 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