127

J. Anat. (1992) 181, pp. 127-131, with 4 figures Printed in Great Britain

Lectin histochemical study of the prostate gland of the rhesus monkey (Macaca mulatta) SHIN WAKUIV, MASAKUNI FURUSATO3, YASUO NOMURA2, MASAO ASARI' AND YUTAKA KANO' Departments of Anatomy' and Pathology', Azabu University School of Veterinary Medicine, Kanagawa, and 'Department of Pathology, Jikei University School of Medicine, Tokyo, Japan (Accepted 16 June 1992)

ABSTRACT

Glycoconjugates in the secretory epithelial cells of the rhesus monkey (Macaca mulatta) prostate gland were investigated using lectin histochemistry. The monkey prostate possesses cranial and caudal lobes. Histochemical staining with a battery of 8 lectins demonstrated differences in lectin binding patterns of the secretory epithelial cells between the 2 lobes. BS-1 bound exclusively to the cranial lobe, and PNA bound exclusively to the caudal lobe. DBA bound exclusively to a number of the secretory epithelial cells of the caudal lobe. In addition, both lobes showed binding with Con-A, RCA-1, WGA and S-WGA, but were negative for binding with SBA. Specific lectin binding patterns suggest a differing carbohydrate composition for each region, and a biological difference between the cranial and caudal lobes of the rhesus monkey prostate.

INTRODUCTION

The monkey prostate gland is located entirely on the dorsal aspect of the urethra and neck of the bladder. It has 2 discrete lobes, cranial and caudal. The cranial lobe has a furrowed surface which closely resembles the external appearance of the seminal vesicle, from which it can easily be separated. The caudal lobe is smooth on its external surface and is located on the dorsolateral aspect of the urethra (Lewis et al. 1981). The cranial and caudal lobes of the monkey prostate have been reported to show different histological and biochemical appearances (Van Wagenen, 1936; Greer et al. 1968; Schoonees et al. 1969; Sufrin et al. 1975; Muntzing et al. 1976; Battersby et al. 1977; Blacklock & Bouskill, 1977; Ghanadian et al. 1977; Karr et al. 1979, 1984; Lewis et al. 1981; De Klerk & Lombard, 1985; Habenicht et al. 1987). Lectins are proteins or glycoproteins that exhibit nonimmune but selective binding to specific carbohydrate moieties within cells and their membranes (Alroy et al. 1984). Using a battery of lectins, several histochemical studies on glycoconjugates related to

the epithelial cells of the prostate have been performed in the dog (Orgad et al. 1984), goat (Tsukise & Yamada, 1984), mouse (Shinha & Bently, 1984), opossum (Nogueira et al. 1985), human (Abel et al. 1987; McNeal et al. 1988), rat (Abel et al. 1987, Hauke et al. 1989), water buffalo (Abou-Elmagd & Wrobel, 1989), pig (Tsukise & Yamada, 1990) and guinea pig (Chan & Wong, 1991). It has been established from these studies that Con-A, RCA-i, WGA and PNA bind similarly to prostatic epithelial cells in these various mammals, but other lectin binding patterns vary considerably from one species to another. For the monkey, however, detailed information is limited to a histochemical analysis of glycoconjugates related to prostatic epithelial cells. The purpose of this study was to define for the rhesus monkey (Macaca mulatta) the differences in secretory epithelial glycoconjugates between the 2 lobes of the prostate using lectin histochemistry.

Correspondence to: Dr Wakui, Department of Anatomy, Azabu University School of Veterinary Medicine, 1-17-71 Fuchinobe, Sagamiharashi, Kamagawa 229, Japan.

128

S. Wakui and others

Table 1. Lectins used for identifying carbohydrate residues in rhesus monkey prostate Concentration

Lectin origin

Common name Abbreviation

(jg/ml)

Major sugar specification*

Binding inhibitor

D. Glca 1 - D. Manor 1Galp 1,4GlcNAci 1-+ NANAa 2,6Galp 1,4GlcNac3 1GlcNAcI 1,4GIcNAc,B 1(Galp 1,4GlcNAco 1-)>2 > sialic acid GlcNAcfi 1,4GlcNAc,B 1-.

a-D-methyl-Man Lactose

Concanavalia ensiformis Jack bean Ricinus communis Castor bean

Con-A RCA-1

10 50

Triticum vulgaris

Wheat germ

WGA

50

Succinyl-WGA

S-WGA

10

Bandeirea simplicifolia Dolichos biflorus

Griffonia Horse gram

BS-1 DBA

50 10

Arachis hypogaea

Peanut

PNA

20

Glycine max

Soyabean

SBA

10

* Gal, galactose;

(Gali 1,4GlcNAcI3 1-)>2 D. Gala 1GalNAca 1,3(Fuca 1,2) Gal 1,4GlcNAc Galf 1,3GalNAca 1 Galp 1,4GlcNAco 1D. GalNAca 1 -. D. Gala 1 GalNAcJ 1-

Sialic acid

-D-GlcNAc Lactose a-D-GalNAc Lactose

a-D-GalNAc

GlcNAc, N-acetyl-glucosamine; GalNAC, N-acetyl-galactosamine; Man, mannose.

MATERIALS AND METHODS

Eleven adult male, intact rhesus monkeys (Macaca mulatta, average weight 6 kg, aged 5-6 y) were used. The animals were housed under standard conditions under the supervision of the Azabu University School of Veterinary Medicine. Samples of the prostate were obtained from animals which had been chemically restrained with ketamine and overdosed with an intravenous injection of pentobarbital sodium. The specimens were fixed with a 10 % solution of buffered formalin for 3-4 d. After dehydration in graded alcohol solutions, the tissues were embedded in paraffin. For histological evaluation, 4 gm sections were cut and stained with haematoxylin and eosin. Lectin histochemistry was performed with biotinylated lectins, avidin-biotin-peroxidase complex and 3,3'-diaminobenzidine tetrahydrochloride-H202 (Alroy et al. 1984). The sections were counterstained with Mayer's haematoxylin for 1 min. Table 1 lists the lectins used in this study, their abbreviations, the lectin concentration used, their major sugar specificities, and the corresponding sugars used to inhibit their tissue binding. We selected those concentrations that give minimal background yet are sufficient to detect relatively low levels of specific sugar residues. Controls for blocking specific lectins included incubating each lectin with a 0.1 M solution of its specific blocking sugar (Table 1) for 20 min before application of the lectin to the tissue sections. RESULTS

The acini in the cranial lobe were lined with a single layer of tall columnar secretory epithelial cells having vacuolated cytoplasm and dense basal nuclei. The

acini in the caudal lobe were lined with a single layer of cuboidal or low columnar secretory epithelial cells having rich secretory granules and vesicular basal nuclei. A few flattened or trilateral basal cells occurred at intervals around the periphery of the acini, interposed between the bases of the epithelial cells. The entire cytoplasm of the secretory epithelial cells of both cranial and caudal lobes was moderately to intensely stained with Con-A, RCA-1, WGA and SWGA (Table 2). Con-A and RCA-1 staining was stronger in the caudal lobe, but WGA and S-WGA stained intensely and equally in both lobes. Con-A and WGA exhibited fine granular staining throughout the entire cytoplasm of the secretory epithelial cells (Fig. 1). RCA-1 and S-WGA showed an accentuated staining intensity towards the luminal plasma membrane (Fig. 2). BS-1 bound selectively to the secretory epithelial cells of the cranial lobe, and PNA bound selectively to those of the caudal lobe (Table 2). BS-1 and PNA exhibited fine granular staining throughout Table 2. Lectin binding sites in secretory epithelial cells of rhesus monkey prostate

Con-A RCA-1 WGA S-WGA BS-1 DBA PNA SBA

Caudal Lobe

Cranial Lobe

++ ++

+ + ++ + +

++ + -

+

-

+

-

-

-

+ +, Intense reaction; +, moderate reaction; -, negative reaction;

+, a few reactive cells.

Lectin histochemistry of monkey prostate

-

.-w, W.,-I,

`

-

_

129

a"

^;

J7

Fig. 1. Secretory epithelial cells of the cranial lobe showing diffuse granular cytoplasmic binding with WGA throughout the cytoplasm. x240.

' -* I'w 4&Ajh...; ;. -Vvq;

t.

-

Fig. 3. Secretory epithelial cells of the cranial lobe showing cytoplasmic binding of BS-1. x 240.

4 i

'-]|1

... .

-_ xeP

,.

_

h.

st

O.. i ;j5:}

F r'

t,t'

Is 4-

_

-

I.

.*

v,

I~~~~~~~~~~~~~~~ ,~~~c a~ o>i s 0

.1

f

"

b

.-

.

..~

M.rb

X

*s,;

jr

,

,

A4

t

. ....

iN fol*'

0.

-p

4%4

.

:9

A

p V

>v

-

s f t> i

-

>bi;

'.4

.q

&ts.^r >p .Z

Fig. 2. Secretory epithelial cells of the caudal lobe showing an accentuation of staining intensity with RCA-I towards the luminal plasma membrane. x 240.

the entire cytoplasm of the secretory epithelial cells (Fig. 3). In addition, DBA bound selectively throughout the cytoplasm of a number of secretory epithelial cells of the caudal lobe, but did not bind the basal cells of either lobe (Table 2, Fig. 4). Both lobes gave negative findings for binding with SBA (Table 2). Lectin binding patterns (except for DBA) for the

Fig. 4. A number of secretory epithelial cells of the caudal lobe showing cytoplasmic binding of DBA. x 240.

prostate cells resembled those of the secretory epithelial cells. DISCUSSION

The rhesus monkey prostate consists of 2 histologically distinct divisions, the cranial and caudal lobes (Battersby et al. 1977; Blacklock & Bouskill, 1977; Ghanadian et al. 1977; Lewis et al. 1981). In the ANA 181

130

S. Wakui and others

present study, the secretory epithelial cells of both lobes bound with Con-A, RCA- 1, WGA and S-WGA, but not with SBA. In addition, specific lectin binding patterns suggested differences in the carbohydrate composition of the secretory epithelial cells of each lobe. a-D-galactosyl residues were present in the cranial lobe, as demonstrated by BS-1 binding. Galp 1,3GalNAca and Galp 1,4GalNAcp residues were present in the caudal lobe, as demonstrated by PNA binding (Pereira et al. 1976). PNA binding patterns in prostatic epithelial cells have been suggested to be dependent on sex hormones. Bischof & Aumuller (1982) have reporited that PNA binding to prostatic epithelial cells in the mature human is high, but low before puberty and in old age. In the dog, PNA binding sites in the prostatic epithelial cells have been removed by castration, and they re-emerge after the administration of androgen and oestrogen (Merk et al. 1986). Cultured rat prostatic epithelial cells which have been exposed to added testosterone show positive PNA binding sites (Martikainen et al. 1986). In the monkey prostate, the cytoplasmic and nuclear androgen receptor levels have been reported to be higher in the caudal lobe as compared with those in the cranial lobe (Sufrin et al. 1975; Ghanadian et al. 1977; Karr et al. 1979, 1984; De Klerk & Lombard, 1985; Habenicht et al. 1987). The cytoplasmic receptor for oestradiol in the caudal lobe is known to exist but not in the cranial lobe of the baboon prostate (Sufrin et al. 1975). The PNA binding pattern of the monkey prostate might also correlate its androgen and oestrogen receptor levels. Structures resembling GalNAca 1,3(Fucal,2)Galp 1,4GlcNAc P1 residues were present in some of the secretory epithelial cells of the caudal lobe, as demonstrated by DBA binding. It might represent unsynchronous secretion by different cells, or different stages of cell renewal or cell degeneration. The differences of DAB binding on the secretory epithelial cells of the caudal lobe might also suggest the possibility of the presence of more than one cell type, as is found in the human prostate (Bischof & Aumuller, 1982), mice (Shinha & Bently, 1984), goat (Tsukise & Yamada, 1984) and water buffalo (Abou-Elmagd & Wrobel, 1989). Sialic acid, identified by positive WGA binding and negative WGA which was incubated with sialic acid before application to the tissue section, was present at the prostatic epithelial cells of both lobes. Sialic acid is known to be a saccharide component which is necessary for the formation and maturation of spermatozoa, and also serves as a source of energy for them (Mann, 1975).

In addition, the 2 lobes of the monkey prostate are distinctive in showing biochemical differences. The greater concentration of radioactive zinc chloride in the caudal lobe is similar to the dorsolateral lobe of the rat and the peripheral zone of the human prostate (Schoonees et al. 1969). Both castration and exogenous testosterone appear to influence the zinc content of the caudal lobe more than that of the cranial lobe (Schoonees et al. 1970). A higher acid phosphatase activity of the epithelial cells was detected in the caudal lobe than in the cranial lobe (Muntzing et al. 1976). In the present study, differences in lectin binding patterns of the secretory epithelial cells between the cranial and caudal lobes of the rhesus monkey prostate indicate different carbohydrate compositions for the secretory epithelial cells of each region. This evidence supports the idea that the monkey prostate gland consists of 2 biologically different organs. REFERENCES

ABEL PD, LEATHEM A, AYLOTT A, MARCH C, HENDERSON D, WILLIAM G (1987) Carbohydrate residues in non-malignant prostatic epithelium as revealed by lectins. Urological Research 15, 173-176. ABOU-ELMAGD A, WROBEL KH (1989) The periurethral glandular complex in the water buffalo: an ultrastructural, histological and

lectin-histochemical study. Archives of Histology and Cytology 52, 501-502. ALROY J, Ucci AA, PEREiRA MEA (1984) Lectins: histochemical probes for specific carbohydrate residues. In Diagnostic Immunochemistry (ed. R. A. DeLellis), pp. 67-88. New York: Masson. BATTERSBY S, CHANDLER JA, HAPER ME, BLACKLOCK NJ (1977)

The ultrastructure of rhesus monkey prostate. Urological Research 5, 175-183. BISCHOF W, AUMULLER G (1982) Age-dependent changes in the carbohydrate pattern of human prostatic epithelium as determined by peroxidase-labeled lectins. Prostate 3, 507-513. BLACKLOCK NJ, BOUSKILL V (1977) The zonal anatomy of the prostate in man and in the rhesus monkey (Macaca mulatta). Urological Research 5, 163-167. CHAN L, WONG YC (1991) Glycoconjugates of the lateral prostate of the guinea-pig: a lectin histochemical study. Prostate 19, 155-172. DE KLERK DP, LOMBARD CJ (1985) Stromal and epithelial growth of the prostate during puberty. Prostate 9, 191-198. GHANADIAN R, SMITH CB, CHISHOLM GS, BLACKLOCK NJ (1977) Differential androgen uptake by the lobes of the rhesus monkey prostate. British Journal of Urology 9, 701-704. GREER WE, ROUSSEL JD, AUSTIN CR (1968) Prevention of coagulation in monkey semen by surgery. Journal ofReproductive Fertility 15, 153-155. HABENICHT UF, ScHwARz K, NEUMANN F, EL-ETREBY MF (1987) Induction of estrogen-related hyperplastic changes in the prostate of the cynomolgus monkey (Macaca fascicularis) by androstenedione and its antagonization by the aromatase inhibitor 1methyl-androsta-1, 4-diene-3, 17-dione. Prostate 11, 313-326. HAUKE CH, HORN R, BREUER W, SINOWATz F (1989) Postnatal development of lectin binding sites in the rat ventral prostate. Histochemical Journal 21, 651-658. KA.RR JP, KIRDANI RY, MURPHY GP, SANDBERG AA (1979) The

131

Lectin histochemistry of monkey prostate baboon prostate as a model for steroid hormone receptors in the human gland. In Prostate Cancer and Hormone Receptors (ed. G. P. Murphy), pp. 165-179. New York: A. R. Liss. KARR JP, CHAI LS, KIM U, MURPHY GP, RESKO JA et al. (1984) Induction of benign prostatic hypertrophy in baboons. Urology 23, 276-289. LEWIS RW, KIM JCS, IRANI D, ROBERTS JA (1981) The prostate of the nonhuman primate: normal anatomy and pathology. Prostate

2, 51-70. MANN T (1975) Biochemistry of semen. In Handbook of Physiology, vol. 5 (ed. D. W. Hamilton & R. 0. Greep), pp. 461-471. Washington: American Physiological Society. McNEAL JE, LEAV I, ALROY J, SHUTELSKY E (1988) Differential lectin staining of central and peripheral zones of the prostate and alterations in dysplasia. American Journal of Clinical Pathology 89, 41-48. MARTIKAINEN P, MALMI R, SUOMINEN J (1986) Distribution of glycoconjugates in normal rat ventral prostate and their use as markers of androgen-controlled secretory function in culture. Prostate 8, 3749. MERK FB, WARHOL MJ, KWAN PW, LEAV I, ALROY J et al. (1986)

Multiple phenotypes of prostatic glandular cells in castrated dogs after individual or combined treatment with androgen and estrogen. Morphometric, ultrastructural, and cytochemical distinctions. Laboratory Investigation 54, 442-456. MUNTZING J, MYHRBERG H, SAROFF T, SANDBERG AA, MURPHY GP (1976) Histochemical and ultrastructural study of prostatic tissue from baboons treated with antiprostatic drug. Investigative Urology 14, 162-167. NOGUEIRA JC, RIBEIRO MG, CAMPOS PA (1985) Histology and

carbohydrate histochemistry of the prostate gland of Brazilian four-eyed opossum (Philander opossum Linnaeus, 1758). Anatomischer Anzeiger 159, 241-252. ORGAD U, ALROY J, Ucci A, MERK FB (1984) Histochemical studies of epithelial cell glycoconjugates in atrophic, metaplastic, hyperplasic, and neoplastic canine prostate. Laboratory Investigation 50, 294-302. PEREIRA MEA, KABAT EA, LOTAN R, SHARON N (1976) Immunohistochemical studies on the specificity of the peanut (Arachis hypogaea) agglutinin. Carbohydrate Research 51, 107118. SCHOONEES R, DE KLERK JN, MURPHY GP (1969) Correlation of prostatic blood flow with zinc activity in intact, castrated and testosterone-treated baboons. Investigative Urology 6, 476-484. SCHOONEES R, DE KLERK JN, MURPHY GP (1970) The effect of prolactin on organ weights and zinc-65 uptake in male baboons. Journal of Surgical Oncology 2, 103. SINHA AA, BENTLY MD (1984) The relationship of epithelial cell types in the ventral prostate glands of castrated mice treated with testosterone. Anatomical Record 208, 533-544. SUFRIN G, KIRDANi RY, SANDBERG AA, MURPHY GP (1975) Estrogen binding and estrogen receptors in the prostate. Surgical Form 26, 584-589. TsuKisE A, YAMADA K (1984) Secretory glycoconjugates in the epithelium of the goat prostate. Histochemical Journal 19, 345-350. TSUKIsE A, YAMADA K (1990) Glycoconjugate histochemistry of the prostate gland in the pig. Acta Histochemica 38, 131-137. VAN WAGENEN G (1936) The coagulating function of the cranial lobe of the prostate gland in the monkey. Anatomical Record 66,

411-421.

Lectin histochemical study of the prostate gland of the rhesus monkey (Macaca mulatta).

Glycoconjugates in the secretory epithelial cells of the rhesus monkey (Macaca mulatta) prostate gland were investigated using lectin histochemistry. ...
1MB Sizes 0 Downloads 0 Views