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Histochemistry of the 3β-hydroxysteroid, 17β-hydroxysteroid and 3α-hydroxysteroid dehydrogenases in human salivary glands
0003-9969;82,070547-05$03.00/O 1982 PergamonPwss Ltd
Archs orul Bid. Vol. 27, pp. 547 to 551, 1982 Printedin Great Britain
HISTOCHEMISTRY OF THE 3P-HYDROXYSTEROID, l’I/bHYDROXYSTEROID AND 3wHYDROXYSTEROID DEHYDROGENASES IN HUMAN SALIVARY GLANDS PAOLA SIRIGU, MARGHERITA Cosu,
*Clinica
MARIA T. PERRA
Istituto di Anatomia Umana Normale Otorinolaringoiatrica, University of Cagliari,
and P.
PUXEDDU*
and 09100 Cagliari,
Italy
Summary-Human parotid and submandibular glands showed no 3cl-hydroxysteroid dehydrogenase (3cr-HSD) activity. The 3/I-hydroxysteroid dehydrogenase (3fi-HSD) and the 17/&hydroxysteroid dehydrogenase (17fl-HSD) appeared intensely reactive in the duct epithelia of the male and female glands and weakly reactive in the acinar cells of the female ones. The failure to demonstrate 3cr-HSD activity indicates that in-uivo androgen activation, if present at all, is not so marked as in target organs. The different distribution of the 3b-HSD and 17fi-HSD in the two sexes can be related not only to the oxidation of androgens but also to the metabolism of the female hormones. Glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) do not seem to be specifically influenced by the sex hormones as their pattern of distribution showed no sex differences.
INTRODUCTION
the 3/I-hydroxysteroid dehydrogenase (3P-HSD; E.C. 1.1.1.51) that of Pearson and Grose (1959) for the 17/I-hydroxysteroid dehydrogenase (17@HSD; EC. 1.1.1.63) and that of Balogh (1966) for the 3a-hydroxysteroid dehydrogenase (3c(-HSD; E.C. 1.1.1.50). The substrates used were dehydroepiandrosterone, testosterone and androsterone respectively, all purchased from the Sigma Chemical Company (St. Louis, MO, U.S.A.). Another group of sections was treated for the demonstration of the G6PD (E.C. 1.1.1.49) and the 6PGD (E.C. 1.1.1.43) (Pearse, 1960). Glucose-6-phosphate disodium salt and 6-phosphogluconate Ba salt were also purchased from Sigma. In each experiment, control sections were incubated with a substrate free medium.
It is known that salivary glands which show sex dimorphism, such as those of mouse, rat and pig, are able to convert the circulating androgens into biologically-active reduced metabolites (Baldi and Charreau, 1972; Coffey, 1973; Booth, 1977). Therefore, they are regarded as androgen target organs. On the other hand, the canine submandibular gland, which is not androgen dependent and shows no sex dimorphism, only oxidizes the androgens (Mosadomi and Ofner, 1976). As human salivary glands do not show any morphological or functional difference related to sex (Scott, 1975; Riva et al., 1976; Testa Riva, 1977) they are not considered as androgen target organs, However, El Attar (1974) and Coffey and Crutchfield (1977) have asserted that the human submandibular gland in vitro metabolizes the androgens through a reductive pathway. In an attempt to reconcile these conflicting data, we have carried out a histochemical investigation of the principal steroid dehydrogenases in human salivary glands. The investigation has been extended to the enzymes glucose-6-phosphate dehydrogenase (G6PD) and 6-phospho-gluconic dehydrogenase (6PGD), both involved in the pentose shunt. This metabolic process seems to be controlled by the sex hormones in some non-genital tissues, such as rodent salivary glands and liver (Nakamura et al., 1974; Teutsch and Rieder, 1979).
RESULTS
The 3/I-HSD and the 17/I-HSD appeared strongly reactive in the duct cells of both parotid and submandibular glands of both sexes, whereas the acinar cells were weakly reactive only in glands from females (Figs l-4). Both acini and ducts were unstained in all the sections treated for 3a-HSD. Sections treated for G6PD and 6PGD showed intense reactivity in the epithelial cells of the duct system, and weak reactivity in the acinar cells. No sex difference was observed (Figs 5-8). DISCUSSION
MATERIALS AND METHODS
The presence of 3p-HSD and 17fi-HSD activities merely shows that steroid metabolism can take place in the duct epithelia of the glands studied, as these enzymes are involved in both reduction and oxidation of the sex hormones. The androgen reduction, which yields biologically active metabolites, requires the presence of See-reductase and 3c(-HSD. Sa-Reductase is not histochemically detectable; 31*-HSD has been demonstrated in some androgen-dependent organs of the rat by Balogh (1966) and man by Sirigu et ul.
Samples of submandibular and parotid glands were obtained from four female and five male subjects, aged 18-50, with cervical .carcinoma. None of the glands showed pathological alterations histologically. The samples were rapidly frozen and, after a few hours, lo-15 pm thick cryostat sections were cut. Some sections were processed for the localization of the steroid dehydrogenases: the method of Wattenberg (1958) was followed for the demonstration of 547
54x
Paola
Sir&
(1981). The entire enzyme system for androgen activation has been biologically detected in several target organs of man and animals (Massa and Martini, 1974; Ofner. Leav and Cavazos, 1974; Djoseland ut al., 1977; Mauvais-Jarvis, 1977) and in particular, in the rodent and porcine salivary glands (Coffey, 1973; Booth. 1977; Coffey, Harvey and Carr, 1979). By contrast, the dog submandibular gland lacks Sa-reductase. but shows 17/&HSD activity (Mosadomi and Ofner, 1976). Our failure to find 3x-HSD activity in human salivary glands does not necessarily mean that it is absent, but could indicate that the enzyme level is too low for the histochemical methods used to show it. Coffey and Crutchfield (1977) have demonstrated 3a-HSD activity in the human submandibular gland. finding androsterone and Sa-androstane 3c(.l7/i-diol as in-vitro metabolites of androstenedione. However. the amount of these metabolites was negligible compared to that produced by the rat submandibular gland (Coffey et al., 1979). As the human submandibular gland has extremely low levels of both Scc-reductase and 3c(-HSD. it cannot be included. in our opinion, among the androgen responsive structures (Massa and Martini, 1974). Therefore, it is a reasonable assumption that the human salivary glands itI t‘ilo chiefly oxidize the circulating androgens. Consequently, they should resemble the canine submandibular gland, rather than porcine and rodent ones. Correspondingly, rodent and porcine, but not human and canine submandibular glands show strong sex dimorphism. Our results do not agree with the conclusions reached by El Attar (1974), who claimed that the reductive pathway of androgen metabolism may occur in the human submandibular gland. However. El Attar has only showed the in-vitro conversion of androstenedione into testosterone due to 17P-HSD activity. In rim. this enzyme could catalyse the inverse reaction as well, so that circulating testosterone is oxidized to form inactive androstenedione. Similarly, the action of 3/?-HSD could be via the oxidation of the circulating dehydroepiandrosterone into androstenedione. Booth, Hay and Dott (1972) histochemically demonstrated sex difference for this enzyme in the pig submandibular gland. In the female pig, the 3/?-HSD activity was confined to the ducts; in the male. it was present in the acini as well. They related this different distribution to a specific androgen action. The same relationship cannot be assumed in the human salivary glands, in which the inverse situation occurs. As 3p-HSD and 17P-HSD show greater diffusion in females than in the male. this difference might be conceivably ascribed to the effects of the female hormones. In fact, not only testosterone. but also progesterone and estrogens reach the saliva (Marder, Joshi and Mandel, 1979; Walker, Read and Riad-Fahmi, 1979; Luisi et d., 1980). Thus. 3b-HSD and 17/3-HSD might be involved in the metabolism of both male and female hormones; consequently, their distribution pattern in the salivary glands might represent the result of complex interactions between all the sex hormones. The presence of GBPD
and 6PGD activities suggests that the pentose shunt, a producer of reduced nicotine adenine dinucleotide phosphate (NADPH)
CI trl
could be functioning. These results are at variance with observations on the rodent submandibular gland and liver, where G6PD reactivity is much stronger in males than in females (Nakamura et al., 1974; Teutsch and Rieder, 1979). In these sites, as in all androgen target tissues, a possible role of the pentose shunt would be the production of the NADPH required for the androger. activation. In androgen-independent organs, such as the human salivary glands, the pentose shunt can be related to other anabolic reactions. including those influenced by the sex hormones rather than to androgen activation. Acknowiedqrmr~lrs This work was supported from the Consiglio Nazionale delle Ricerche. thank Mr A. Cadau for his valuable technical
by a grant We wish to assistance.
REFERENCES Baldi A. and Charreau E. H. 1972. Testosterone metabolism by rodent submaxillary glands m relation to their sexual dimorphism. ANU phrsiol. latinoam. 22, 129-134. Balogh K. 1966. Histochemical demonstration of 3r-hydroxysteroid dehydrogenase activity. J. Hisfochem Cytor~hrm. 14, 77-83. Booth W. D. 1977. Metabolism of androgens in vitro by the submaxillary salivary gland of the mature domestic boar. J. Endnrr. 75. 145-154. Booth W. D.. Hay M. F. and Dott H. M. 1972. Sexual dimorphism in the submaxillary gland of the pig. J. Rrprod. Fert. 33, 163-166. Coffey J. C. 1973. Steroid metabolism by mouse submaxillary glands-I. In citro metabolism of testosterone and 4-androstene-3,17-dione. Strroids 22, 247-257. Coffey J. C. and Crutchfield W. 1977. In ritro metabolism
of 4-androstene-3.17-dione by human submaxillary gland homogenates. J. dmt. Rrs. 56, 332-334. Coffey J. C., Harvey T. E. and Carr W. L. 1979. In vitro metabolism of 4-androstene-3,17-dione and testosterone by rat submaxillary gland. Strroids 33, 223-232. Djr-lseland 0.. Tveter K. J., Attramadal A.. Hansson V.. Haugen H. N. and Mathisen W. 1977. Metabolism of testosterone in the human prostate and seminal vesicle. Scund. J. urol. Nephrol. II, I 6. El Attar T. M. A. 1974. In vitro metabolism of estrone 2.4,6.7-‘H and 4-androstene-3.1 7-dione-1,2-3H in submandibular gland and submandibular gland cancer tumor. Steroids 24, 519-526. Luisi M.. Bernini G. P., Del Genovese A.. Birindellr R.. Barletta D., Gasperi M. and Franchi F. 1980. Radioimmunoassay for “free” testosterone in human saliva. J. Stwoid Biochrm. 12, 5 13- 5 16. Marder M. Z., Joshi U. and Mandel 1. D. 1979. Estrogen concentration in human parotid and submaxillary saliva. J. dent. Rrs. 58, 2370.
Massa R. and Martini L. 1974. Testosterone metabolism: necessary step for activity’? J. Strroid Biochem.
a 5,
941~ 941.
Mauvais-Jarvis P. 1977. Androgen metabolism in human skin: mechanism of control. In: Androyens und Antiundroyen.~ (Edited by Martini L. and Motta M.) pp. 229-243. Raven Press, New York. Mosadomi A. and Ofner P. 1976. In vitro metabolism of testosterone-4-‘4C by canine salivary glands. J. oral Puth. 5, 33-41. Nakamura T.. Fujii M.. Kalho M. and Kumegawa M. 1974. Sex difference in glucose-6.phosphate dehydrogenase activity in the submandibular gland of mice. Bicjc&m. hiophys. Acta 362, 1 lo--120. Ofner P.. Leav 1. and Cavazos L. F. 1974. Mu/r Accussor!, SL’Y Oryuns. Strtrcturr and Function in Mammals (Edited
Steroid
dehydrogenases
by Brandes D.) Chap. 11, p. 274. Academic Press, New York. Pearse A. G. E. 1960. Histochemistry, Throrrtical and Applied. Churchill, London. Pearson B. and Grose F. 1959. Histochemical demonstration of 17fl-hydroxysteroid dehydrogenase by use of tetrazolium salt. &oc. &. exp. Bioi. Med. 100, 636638. Riva A., Testa Riva F.. Del Fiacco M. and Lantini M. S. 1976. Fine structure and cytochemistry of the intralobular ducts of the human parotid gland. J. Anat. 122, 627-640. Scott J. 1975. Age, sex and contralateral differences in the volumes of human submandibular salivary glands. Archs oral Biol. 20, 8X5-887. Sirigu P., Cossu M.. Scarpa R. and Pinna A. 1981. Histochemistry of some steroid dehydrogenases in epithelia of
Plate
in human
glands
549
human seminal vesicle, deferential ampulla and prostate gland. Arcl~s And& 7, 9-14. Testa Riva F. 1977. Ultrastructure of human submandibular aland. J. suhmicr. C\TO/.9. 251-266. Teutsih H. F. and Riede; H. 1979. NADP-dependent dehydrogenases in rat liver parenchymaII. Comparison of qualitative and quantitative G6PDH distribution patterns with particular reference to sex differences. Histochemistry 60, 43-52. Walker R. F.. Read G. F. and Riad-Fahmi D. 1979. Radioimmunoassay of progesterone in saliva: application to the assessment of ovarian function. C/in. Chum. 25, 2030-2033. Wattenberg L. W. 1958. Microscopic histochemical demonstration of steroid-3fl-ol dehydrogenase in tissue sections. J. Hisrochrm. Cytochem. 6, 225 232.
1 overleaf.
Paola
550
Smgu
Plate Fig. 1. Female
submandibular
Fig. 2. Male submandibular Fig. 3. Female
parotid
I.
gland:
the 3/1-HSD activtty acini. x 203
appears
to be localized
gland:
the 3p-HSD activity
is confined
Fig. 4. Male parotid
Fig. 6. Male submandibular parotid
gland:
17fi-HSD
activity
gland:
Fig. 8. Male parotid
G6PD
distribution is identical x 203
the 6PGD gland:
6PGD
reactivity activity.
as 3&HSD, x 203
is only in the ducts.
gland: the G6PD appears to be strongly weakly reactive in the acini. x 82 gland:
in both
to the duct epithelium.
gland: 17/LHSD activity shows the same distribution reacttve in the ducts and weakly reactive in the acini.
Fig. 5. Female submandibular
Fig. 7. Female
rt (II.
reactive
ducts
and
x 335
being strongly
x 335 in the ducts
to that observed
and mare
in the female gland.
shows a localization
Identical
No sex difference
is detectable.
to G6PD. x 335
x 335
Steroid dehydrogenases in human glands
i
Plate 1.
551
Archs orul Bid. Vol. 27, pp. 547 to 551, 1982 Printedin Great Britain
HISTOCHEMISTRY OF THE 3P-HYDROXYSTEROID, l’I/bHYDROXYSTEROID AND 3wHYDROXYSTEROID DEHYDROGENASES IN HUMAN SALIVARY GLANDS PAOLA SIRIGU, MARGHERITA Cosu,
*Clinica
MARIA T. PERRA
Istituto di Anatomia Umana Normale Otorinolaringoiatrica, University of Cagliari,
and P.
PUXEDDU*
and 09100 Cagliari,
Italy
Summary-Human parotid and submandibular glands showed no 3cl-hydroxysteroid dehydrogenase (3cr-HSD) activity. The 3/I-hydroxysteroid dehydrogenase (3fi-HSD) and the 17/&hydroxysteroid dehydrogenase (17fl-HSD) appeared intensely reactive in the duct epithelia of the male and female glands and weakly reactive in the acinar cells of the female ones. The failure to demonstrate 3cr-HSD activity indicates that in-uivo androgen activation, if present at all, is not so marked as in target organs. The different distribution of the 3b-HSD and 17fi-HSD in the two sexes can be related not only to the oxidation of androgens but also to the metabolism of the female hormones. Glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) do not seem to be specifically influenced by the sex hormones as their pattern of distribution showed no sex differences.
INTRODUCTION
the 3/I-hydroxysteroid dehydrogenase (3P-HSD; E.C. 1.1.1.51) that of Pearson and Grose (1959) for the 17/I-hydroxysteroid dehydrogenase (17@HSD; EC. 1.1.1.63) and that of Balogh (1966) for the 3a-hydroxysteroid dehydrogenase (3c(-HSD; E.C. 1.1.1.50). The substrates used were dehydroepiandrosterone, testosterone and androsterone respectively, all purchased from the Sigma Chemical Company (St. Louis, MO, U.S.A.). Another group of sections was treated for the demonstration of the G6PD (E.C. 1.1.1.49) and the 6PGD (E.C. 1.1.1.43) (Pearse, 1960). Glucose-6-phosphate disodium salt and 6-phosphogluconate Ba salt were also purchased from Sigma. In each experiment, control sections were incubated with a substrate free medium.
It is known that salivary glands which show sex dimorphism, such as those of mouse, rat and pig, are able to convert the circulating androgens into biologically-active reduced metabolites (Baldi and Charreau, 1972; Coffey, 1973; Booth, 1977). Therefore, they are regarded as androgen target organs. On the other hand, the canine submandibular gland, which is not androgen dependent and shows no sex dimorphism, only oxidizes the androgens (Mosadomi and Ofner, 1976). As human salivary glands do not show any morphological or functional difference related to sex (Scott, 1975; Riva et al., 1976; Testa Riva, 1977) they are not considered as androgen target organs, However, El Attar (1974) and Coffey and Crutchfield (1977) have asserted that the human submandibular gland in vitro metabolizes the androgens through a reductive pathway. In an attempt to reconcile these conflicting data, we have carried out a histochemical investigation of the principal steroid dehydrogenases in human salivary glands. The investigation has been extended to the enzymes glucose-6-phosphate dehydrogenase (G6PD) and 6-phospho-gluconic dehydrogenase (6PGD), both involved in the pentose shunt. This metabolic process seems to be controlled by the sex hormones in some non-genital tissues, such as rodent salivary glands and liver (Nakamura et al., 1974; Teutsch and Rieder, 1979).
RESULTS
The 3/I-HSD and the 17/I-HSD appeared strongly reactive in the duct cells of both parotid and submandibular glands of both sexes, whereas the acinar cells were weakly reactive only in glands from females (Figs l-4). Both acini and ducts were unstained in all the sections treated for 3a-HSD. Sections treated for G6PD and 6PGD showed intense reactivity in the epithelial cells of the duct system, and weak reactivity in the acinar cells. No sex difference was observed (Figs 5-8). DISCUSSION
MATERIALS AND METHODS
The presence of 3p-HSD and 17fi-HSD activities merely shows that steroid metabolism can take place in the duct epithelia of the glands studied, as these enzymes are involved in both reduction and oxidation of the sex hormones. The androgen reduction, which yields biologically active metabolites, requires the presence of See-reductase and 3c(-HSD. Sa-Reductase is not histochemically detectable; 31*-HSD has been demonstrated in some androgen-dependent organs of the rat by Balogh (1966) and man by Sirigu et ul.
Samples of submandibular and parotid glands were obtained from four female and five male subjects, aged 18-50, with cervical .carcinoma. None of the glands showed pathological alterations histologically. The samples were rapidly frozen and, after a few hours, lo-15 pm thick cryostat sections were cut. Some sections were processed for the localization of the steroid dehydrogenases: the method of Wattenberg (1958) was followed for the demonstration of 547
54x
Paola
Sir&
(1981). The entire enzyme system for androgen activation has been biologically detected in several target organs of man and animals (Massa and Martini, 1974; Ofner. Leav and Cavazos, 1974; Djoseland ut al., 1977; Mauvais-Jarvis, 1977) and in particular, in the rodent and porcine salivary glands (Coffey, 1973; Booth. 1977; Coffey, Harvey and Carr, 1979). By contrast, the dog submandibular gland lacks Sa-reductase. but shows 17/&HSD activity (Mosadomi and Ofner, 1976). Our failure to find 3x-HSD activity in human salivary glands does not necessarily mean that it is absent, but could indicate that the enzyme level is too low for the histochemical methods used to show it. Coffey and Crutchfield (1977) have demonstrated 3a-HSD activity in the human submandibular gland. finding androsterone and Sa-androstane 3c(.l7/i-diol as in-vitro metabolites of androstenedione. However. the amount of these metabolites was negligible compared to that produced by the rat submandibular gland (Coffey et al., 1979). As the human submandibular gland has extremely low levels of both Scc-reductase and 3c(-HSD. it cannot be included. in our opinion, among the androgen responsive structures (Massa and Martini, 1974). Therefore, it is a reasonable assumption that the human salivary glands itI t‘ilo chiefly oxidize the circulating androgens. Consequently, they should resemble the canine submandibular gland, rather than porcine and rodent ones. Correspondingly, rodent and porcine, but not human and canine submandibular glands show strong sex dimorphism. Our results do not agree with the conclusions reached by El Attar (1974), who claimed that the reductive pathway of androgen metabolism may occur in the human submandibular gland. However. El Attar has only showed the in-vitro conversion of androstenedione into testosterone due to 17P-HSD activity. In rim. this enzyme could catalyse the inverse reaction as well, so that circulating testosterone is oxidized to form inactive androstenedione. Similarly, the action of 3/?-HSD could be via the oxidation of the circulating dehydroepiandrosterone into androstenedione. Booth, Hay and Dott (1972) histochemically demonstrated sex difference for this enzyme in the pig submandibular gland. In the female pig, the 3/?-HSD activity was confined to the ducts; in the male. it was present in the acini as well. They related this different distribution to a specific androgen action. The same relationship cannot be assumed in the human salivary glands, in which the inverse situation occurs. As 3p-HSD and 17P-HSD show greater diffusion in females than in the male. this difference might be conceivably ascribed to the effects of the female hormones. In fact, not only testosterone. but also progesterone and estrogens reach the saliva (Marder, Joshi and Mandel, 1979; Walker, Read and Riad-Fahmi, 1979; Luisi et d., 1980). Thus. 3b-HSD and 17/3-HSD might be involved in the metabolism of both male and female hormones; consequently, their distribution pattern in the salivary glands might represent the result of complex interactions between all the sex hormones. The presence of GBPD
and 6PGD activities suggests that the pentose shunt, a producer of reduced nicotine adenine dinucleotide phosphate (NADPH)
CI trl
could be functioning. These results are at variance with observations on the rodent submandibular gland and liver, where G6PD reactivity is much stronger in males than in females (Nakamura et al., 1974; Teutsch and Rieder, 1979). In these sites, as in all androgen target tissues, a possible role of the pentose shunt would be the production of the NADPH required for the androger. activation. In androgen-independent organs, such as the human salivary glands, the pentose shunt can be related to other anabolic reactions. including those influenced by the sex hormones rather than to androgen activation. Acknowiedqrmr~lrs This work was supported from the Consiglio Nazionale delle Ricerche. thank Mr A. Cadau for his valuable technical
by a grant We wish to assistance.
REFERENCES Baldi A. and Charreau E. H. 1972. Testosterone metabolism by rodent submaxillary glands m relation to their sexual dimorphism. ANU phrsiol. latinoam. 22, 129-134. Balogh K. 1966. Histochemical demonstration of 3r-hydroxysteroid dehydrogenase activity. J. Hisfochem Cytor~hrm. 14, 77-83. Booth W. D. 1977. Metabolism of androgens in vitro by the submaxillary salivary gland of the mature domestic boar. J. Endnrr. 75. 145-154. Booth W. D.. Hay M. F. and Dott H. M. 1972. Sexual dimorphism in the submaxillary gland of the pig. J. Rrprod. Fert. 33, 163-166. Coffey J. C. 1973. Steroid metabolism by mouse submaxillary glands-I. In citro metabolism of testosterone and 4-androstene-3,17-dione. Strroids 22, 247-257. Coffey J. C. and Crutchfield W. 1977. In ritro metabolism
of 4-androstene-3.17-dione by human submaxillary gland homogenates. J. dmt. Rrs. 56, 332-334. Coffey J. C., Harvey T. E. and Carr W. L. 1979. In vitro metabolism of 4-androstene-3,17-dione and testosterone by rat submaxillary gland. Strroids 33, 223-232. Djr-lseland 0.. Tveter K. J., Attramadal A.. Hansson V.. Haugen H. N. and Mathisen W. 1977. Metabolism of testosterone in the human prostate and seminal vesicle. Scund. J. urol. Nephrol. II, I 6. El Attar T. M. A. 1974. In vitro metabolism of estrone 2.4,6.7-‘H and 4-androstene-3.1 7-dione-1,2-3H in submandibular gland and submandibular gland cancer tumor. Steroids 24, 519-526. Luisi M.. Bernini G. P., Del Genovese A.. Birindellr R.. Barletta D., Gasperi M. and Franchi F. 1980. Radioimmunoassay for “free” testosterone in human saliva. J. Stwoid Biochrm. 12, 5 13- 5 16. Marder M. Z., Joshi U. and Mandel 1. D. 1979. Estrogen concentration in human parotid and submaxillary saliva. J. dent. Rrs. 58, 2370.
Massa R. and Martini L. 1974. Testosterone metabolism: necessary step for activity’? J. Strroid Biochem.
a 5,
941~ 941.
Mauvais-Jarvis P. 1977. Androgen metabolism in human skin: mechanism of control. In: Androyens und Antiundroyen.~ (Edited by Martini L. and Motta M.) pp. 229-243. Raven Press, New York. Mosadomi A. and Ofner P. 1976. In vitro metabolism of testosterone-4-‘4C by canine salivary glands. J. oral Puth. 5, 33-41. Nakamura T.. Fujii M.. Kalho M. and Kumegawa M. 1974. Sex difference in glucose-6.phosphate dehydrogenase activity in the submandibular gland of mice. Bicjc&m. hiophys. Acta 362, 1 lo--120. Ofner P.. Leav 1. and Cavazos L. F. 1974. Mu/r Accussor!, SL’Y Oryuns. Strtrcturr and Function in Mammals (Edited
Steroid
dehydrogenases
by Brandes D.) Chap. 11, p. 274. Academic Press, New York. Pearse A. G. E. 1960. Histochemistry, Throrrtical and Applied. Churchill, London. Pearson B. and Grose F. 1959. Histochemical demonstration of 17fl-hydroxysteroid dehydrogenase by use of tetrazolium salt. &oc. &. exp. Bioi. Med. 100, 636638. Riva A., Testa Riva F.. Del Fiacco M. and Lantini M. S. 1976. Fine structure and cytochemistry of the intralobular ducts of the human parotid gland. J. Anat. 122, 627-640. Scott J. 1975. Age, sex and contralateral differences in the volumes of human submandibular salivary glands. Archs oral Biol. 20, 8X5-887. Sirigu P., Cossu M.. Scarpa R. and Pinna A. 1981. Histochemistry of some steroid dehydrogenases in epithelia of
Plate
in human
glands
549
human seminal vesicle, deferential ampulla and prostate gland. Arcl~s And& 7, 9-14. Testa Riva F. 1977. Ultrastructure of human submandibular aland. J. suhmicr. C\TO/.9. 251-266. Teutsih H. F. and Riede; H. 1979. NADP-dependent dehydrogenases in rat liver parenchymaII. Comparison of qualitative and quantitative G6PDH distribution patterns with particular reference to sex differences. Histochemistry 60, 43-52. Walker R. F.. Read G. F. and Riad-Fahmi D. 1979. Radioimmunoassay of progesterone in saliva: application to the assessment of ovarian function. C/in. Chum. 25, 2030-2033. Wattenberg L. W. 1958. Microscopic histochemical demonstration of steroid-3fl-ol dehydrogenase in tissue sections. J. Hisrochrm. Cytochem. 6, 225 232.
1 overleaf.
Paola
550
Smgu
Plate Fig. 1. Female
submandibular
Fig. 2. Male submandibular Fig. 3. Female
parotid
I.
gland:
the 3/1-HSD activtty acini. x 203
appears
to be localized
gland:
the 3p-HSD activity
is confined
Fig. 4. Male parotid
Fig. 6. Male submandibular parotid
gland:
17fi-HSD
activity
gland:
Fig. 8. Male parotid
G6PD
distribution is identical x 203
the 6PGD gland:
6PGD
reactivity activity.
as 3&HSD, x 203
is only in the ducts.
gland: the G6PD appears to be strongly weakly reactive in the acini. x 82 gland:
in both
to the duct epithelium.
gland: 17/LHSD activity shows the same distribution reacttve in the ducts and weakly reactive in the acini.
Fig. 5. Female submandibular
Fig. 7. Female
rt (II.
reactive
ducts
and
x 335
being strongly
x 335 in the ducts
to that observed
and mare
in the female gland.
shows a localization
Identical
No sex difference
is detectable.
to G6PD. x 335
x 335
Steroid dehydrogenases in human glands
i
Plate 1.
551