Spironolactone bodies in an adrenal adenoma.

Spironolactone bodies were observed in an adrenal cortical adenoma that was removed from a patient with primary hyperaldosteronism (Conn syndrome) treated preoperatively with spironolactone. The electron microscopical evaluation of this adrenal cortical adenoma shows origin of spironolactone bodies from whorls of endoplasmic reticulum in cells with the cytoplasmic features of those from the zona glomerulosa of the adrenal gland. There was no evidence that the bodies were derived from mitochondria, which confirms recent ultrastructural findings in patients treated with spironolactone. These bodies have been described in the adrenal cortex only in patients who have received spironolactone, and the pharmacologic specificity of the bodies strongly suggests a direct mode of action by spironolactone on aldosterone production by cells of the adrenal zona glomerulosa.     

Factors affecting the frequency of occurrence of spironolactone bodies in aldosteronomas and non-tumorous cortex

Histological sections taken from aldosterone-producing-adenomas and from non-tumorous adrenal cortex of 18 patients treated for primary aldosteronism by unilateral adrenalectomy were examined for spironolactone inclusion bodies. Typical inclusions were found in 10 of the 13 patients who received spironolactone up to within 24 h of surgery. They were present in the tumours of 7 patients, and their frequency correlated positively with the percentage of glomerulosa type cells in the tumours. In tumours containing 50% or more glomerulosa-type cells, their frequency correlated negatively with duration of treatment. They were present in the non-tumorous cortex of 4 of these 7 patients, and in the cortex of 3 others whose tumours did not contain them. In the cortex, they were found only in glomerulosa cells, and their presence appeared unrelated to dosage or duration of treatment. No spironolactone inclusion bodies were seen in either the tumour of the non-tumorous cortex of 3 patients who had discontinued spironolactone 19 to 97 d before surgery, or of 2 patients who had never received spironolactone.     

Spironolactone bodies in aldosteronomas and in the attached adrenals. Enzyme histochemical study of 19 cases of primary aldosteronism and a case of aldosteronism due to bilateral diffuse hyperplasia of the zona glomerulosa.

The formation of spironolactone (S) bodies, eosinophilic laminated cytoplasmic inclusions, is induced in the aldosterone-producing cells of the human adrenal cortex after the administration of spironolactone. The aim of this study was to define the enzyme histochemical characteristics of S bodies, S-body-containing cells, and the apparently hyperplastic zona glomerulosa (zG) of adrenal tissues attached to aldosteronomas. S bodies were found in 14 of 19 aldosteronomas, in 10 of 19 adrenal tissues attached to aldosteronomas, and in the adrenal tissues in a patient with aldosteronism due to bilateral diffuse zG hyperplasia. The S bodies themselves exhibited most intense 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) activity but did not exhibit glucose-6-phosphate dehydrogenase (G6PD), NADP-dependent isocitrate dehydrogenase (NADP-ICDH), or succinate dehydrogenase (SDH) activity, confirming histochemically the origin of S bodies in the smooth endoplasmic reticulum. In two adenomas, S bodies were found to be surrounded by reaction products of acid hydrolase but were not found in the other adenomas and the remaining adrenal tissues. S-body-containing cells, irrespective of being neoplastic or not, showed enhanced 3 beta HSD, G6PD, and NADP-ICDH activity and weak SDH activity (Type I pattern of enzyme activity). Though zG was hyperplastic in most of the adrenal tissues attached to the adenomas, zG cells that did not contain S bodies showed the opposite pattern (Type II pattern) of enzyme activity (ie, weak 3 beta HSD, G6PD, and NADP-ICDH activity and intense SDH activity), in contrast to those in the adrenal tissues in a patient with aldosteronism due to bilateral diffuse zG hyperplasia (which showed the Type I pattern). The results are consistent with the view that hyperplastic zG cells, except S-body-containing cells, in the case of aldosteronoma are not hyperfunctioning. The latter cells may have enhanced but possibly abortive steroidogenic activity.     

Quantitative ultrastructural study of the rat adrenal cortex in renal encapsulation-induced hypertension.

Hypertension was induced in young rats by latex encapsulation of both kidneys. By the fourth week, 85% of the renal-encapsulated (RE) rats became hypertensive. Varying degrees of cardiovascular involvement were evident in the moderately to severely hypertensive rats. The level of systolic blood pressure was directly correlated with the width and the volume of zona glomerulosa of the adrenal cortex. Electron microscopy combined with morphometric-stereologic techniques was employed to quantitate change in the adrenal cortex. The cells of both zona glomerulosa and zona fasciculata of RE rats showed significant increases in the volume of the cell, nucleus, smooth endoplasmic reticulum, and lipid droplets; only in the zona glomerulosa cells was the increase in surface area of the smooth endoplasmic reticulum statistically significant. It is suggested that these structural changes associated with renal-encapsulation hypertension are related at least in part to stress of the hypertensive cardiovascular disease.     

Effects of glucocorticoids on differentiation of zona glomerulosa of fetal adrenal cortex of rats.

The tissue differentiation of the zona glomerulosa of the fetal adrenal cortex of rats was studied by giving experimental treatments to the fetus in vivo. A low-glucocorticoid-condition was given to the fetus by bilateral adrenalectomy of pregnant rats for removing exogenous glucocorticoids from the fetus, and by brain aspiration of the fetuses for removing the fetal pituitary gland (ACTH) and endogenous glucocorticoids. When the fetus was placed under a low-glucocorticoid-condition for the last couple of days of gestation, poor differentiation of the zona glomerulosa occurred specifically in the fetal adrenal cortex. The degree of the poor differentiation seemed to be proportional to the duration of the low-glucocorticoid-condition. Supplemental administration of glucocorticoids could prevent this poor differentiation of the zona glomerulosa. These results indicate that the tissue differentiation of the zona glomerulosa of the fetal adrenal cortex depends much on glucocorticoids.     

HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES OF THE ADRENAL CORTEX IN SPONTANEOUSLY HYPERTENSIVE RATS WITH HIGH SALT SOLUTION

The response of the adrenal cortex In weanling Spontaneously Hypertensive Rats (SHR, OKAMOTO and AOKI) and Wistar-Kyoto Rats (WKR) when given 1% NaCl In drinking water was studied histochemically and electron microscopically. The development of hypertension was accelerated in SHR given 1% NaCl in drinking water. The thickness of the zona glomerulosa In WKR receiving 1% NaCl in drinking water remarkably diminished following the course of experiment and lipid droplets In the zona glomerulosa decreased gradually. The zona glomerulosa cells revealed shrinkage of the cells and a decreased number of mitochondrial crlstae were observed at three weeks of experiment. The remaining tubular crlstae of mitochondria possessed a milled outline. Golgi apparatus became small and fragmental. Membrane -bounded dense bodies were prominent in the peripheral portion of the cell and around Golgl apparatus. In SHR given 1% NaCl in drinking water, the width of the zona glomerulosa less diminished. A marked decrease in number and size of lipid droplets was seen. However, there was no distinct alteration of cytoplasmic organelles. These morphological changes were discussed from the standpoint of functional state In the zona glomerulosa with relation to the development of hypertension in SHR.     

EFFECTS OF GLUCOCORTICOIDS ON DIFFERENTIATION OF ZONA GLOMERULOSA OF FETAL ADRENAL CORTEX OF RATS

The tissue differentiation of the zona glomerulosa of the fetal adrenal cortex of rats was studied by giving experimental treatments to the fetus in vivo. A low-glucocorticoid-condition was given to the fetus by bilateral adrenalectomy of pregnant rats for removing exogenous glucocorticoids from the fetus, and by brain aspiration of the fetuses for removing the fetal pituitary gland (ACTH) and endogenous glucocorticoids. When the fetus was placed under a low-glucocorticoid-condition for the last couple of days of gestation, poor differentiation of the zona glomerulosa occurred speciflcally in the fetal adrenal cortex. The degree of the poor differentiation seemed to be proportional to the duration of the low-glucocorticoid-condition. Supplemental administration of glucocorticoids could prevent this poor differentiation of the zona glomerulosa. These results indicate that the tissue differentiation of the zona glomerulosa of the fetal adrenal cortex depends much on glucocorticoids.     

Ultrastructural studies of adrenal adenoma causing primary aldosteronism

Summary Adrenal adenoma tissue was obtained from 7 patients with the diagnosis of primary aldosteronism and was studied electron microscopically. Spironolactone was administered in 6 of these patients, but not in the remaining patient. Most of the mitochondria of the tumour cells possessed tubular cristae, giving an appearance similar to the mitochondria in the cells of the zona glomerulosa. Spironolactone bodies were seen in the tumour cells of 6 patients who were given spironolactone preoperatively, but were not observed in these cells in the patient not given spironolactone. The literature on the developmental mechanism of this spironolactone body was reviewed.     

Effects of the hypolipidemic drug nafenopin on the zona glomerulosa of the rat adrenal cortex: morphological counterparts of functional alterations

The effects of nafenopin, a hypolipidemic drug, on the zona glomerulosa of the rat adrenal cortex were investigated. Chronic nafenopin treatment significantly lowered serum cholesterol level, but did not alter blood aldosterone concentration, though the biosynthesis of adrenal cortico-steroid hormones seems to be largely dependent upon a continuous uptake of cholesterol from plasma lipoproteins. Stereology showed that the treatment provoked a notable lipid droplet depletion, coupled with a significant proliferation of smooth endoplasmic reticulum (SER) profiles. Since SER is known to be involved in the endogenous synthesis of cholesterol, the hypothesis is advanced that SER hypertrophy is a compensatory response enabling zona glomerulosa cells to maintain an adequate level of aldosterone output even in the absence of a normal supply of exogenous cholesterol.     

Distribution of prothymosin alpha in rat and human adrenal cortex

Background: Prothymosin alpha (ProT) is a polypeptide widely distributed in the organism and expressed by cell types with a high proliferative capacity. The aim of the current work was to investigate if ProT was localized in the progenitor compartment of the adrenal cortex which, following the cell migration theory, corresponds to the zona glomerulosa. Methods: We studied by immunohistochemical and in situ hybridization methods the distribution of ProT in rat and human adrenal cortex. Immunohistochemical techniques for the study of the proliferating cell nuclear antigen (PCNA) and incorporation of 5-bromo-2′-deoxyuridin during DNA synthesis were also done. Immunoelectron microscopic procedures were performed to determine the exact subcellular localization of ProT. Results: ProT was found in the zona glomerulosa cells, but not in the cells of the remaining cortical layers (zonae fasciculata and reticularis). Glomerulosa cells showed immunostaining for ProT only in the nuclei, excluding the nucleoli. Variability in immunostaining intensity was found between different glomerulosa cells. In situ hybridization of ProT mRNA confirmed that ProT synthesis in adrenal cortex occurs only in the zona glomerulosa. The results obtained for proliferating cell nuclear antigen and incorporation of 5-bromo-2′-deoxyuridin confirmed that adrenocortical proliferation occurs in the zona glomerculosa. Ultrastructural immunocytochemistry showed labelling for ProT over the euchromatin, but not on the heterochromatin aggregations nor the nucleoli. Conclusions: The results presented here: (1) support the migration theory for the adrenocortical cell renewal, (2) demonstrate that ProT is present in the nuclei of proliferating cells (being associated with euchromatin), and (3) suggest that the study of ProT expression would be useful in distinguishing cycling from resting cells. © 1994 Wiley-Liss, Inc.     

The adrenal cortex in spontaneously hypertensive rats. A quantitative ultrastructural study.

The adrenal cortex of spontaneously hypertensive rats (SHR) has been examined by quantitative morphologic techniques for electron microscopy. The volume and surface area of smooth endoplasmic reticulum and the volume of Golgi apparatuses in zona glomerulosa cells of SHR was significantly greater than those of Wistar-Kyoto strain (W/KY) normotensive controls; the volume of lipid droplets and nucleus was significantly less in SHR than in W/KY animals. A stimulation of the zona glomerulosa in SHR may well be attributable to the elevation in systolic blood pressure. A distinct lipid-free subglomerulosa was observed in the adrenal gland of W/KY rats; the cell volume was similar to that of the zona glomerulosa although the cells showed a significantly greater volume of mitochondria and surface area of mitochondrial membranes and greater volume of smooth endoplasmic reticulum and lysosomes. In the zona fasciculata, cell volume, volumes and surface area of mitochondria and smooth endoplasmic reticulum, and volume of lipid droplets were significantly lower in SHR than in W/KY rats. The volume of the Golgi apparatus was greater in SHR than in W/KY rats. Glycogen particles were observed in focal areas of some zona fasciculata cells. The adrenal cortex of another strain of normotensive Wistar rat (W/CFN) was compared with that of the W/KY and SHR. Although the relative adrenal weights of SHR and W/KY animals were identical, the weight of that in W/CFN was significantly smaller. The volume of the zona glomerulosa of SHR was significantly greater than that of W/KY although the volume of the zona glomerulosa in W/CFN was significantly greater than the other two groups. The volume of nucleus and lipid droplets of zona glomerulosa in W/KY was significantly greater than that in the S/CFN; the volume of the cell, mitochondria, smooth endoplasmic reticulum, lipid droplets, and lysosomes, and the surface area of smooth endoplasmic reticulum and mitochondrial membranes of W/KY animals was significantly greater than those of W/CFN animals. It is concluded that the W/CFN rat is not an appropriate control for spontaneously hypertensive rats.     

Structure and dynamics of adrenal mitochondria following stimulation with corticotropin releasing hormone

The mitochondria of rat adrenals were investigated qualitatively and quantitatively in different functional states of the adrenal cortex. Following stimulation of the animals with corticotropin releasing hormone (CRH), the corticosterone serum levels reached a maximum 1 hour after stimulation with CRH. The amount of inner mitochondrial membrane within the zona fasciculata increased showing a biphasic time course, with a first maximum 2 hours and a second maximum 8 hours after stimulation. In contrast, a significant rise of mitochondrial volume occurred only 24 hours after CRH stimulation. Therefore, the dense vesicularization of mitochondrial cristae may constitute an early process to enhance the steroidogenic capacity of these cells. Within cells of the transition zone between zona glomerulosa and zona fasciculata, we could depict a special type of mitochondria with characteristic crescent-like cristae only seen after stimulation with CRH. This type of mitochondria may represent an intermediate form between mitochondria of zona glomerulosa and zona fasciculata underlining the impressive transformational capacity of adrenocortical mitochondria. After hypophysectomy, zona fasciculata cells contained mitochondria with tubular inner membranes, representing a hypofunctional state. In contrast, the hypofunctional state after hypophysectomy and the hyperfunctional state after stimulation of the adrenal cortex via CRH injection did not appear to correlate with the morphology of mitochondria from the zona reticularis and adrenal medulla.© Willey-Liss, Inc.     

Interrenal cell zonation in the adrenal gland of the goose (Anser anser)

The adrenal medulla in mammalian species is surrounded by a cortex that contains three distinct layers, whereas the cortex and medulla are intermingled in poultry species. The objective of the present study was to determine the distinct zonation changes in the adrenal cortex of geese in various ages using both electron and light microscopy. Adrenal glands were obtained from French Geese (Anser anser) under deep ether anesthesia at posthach day 1, 5, 10, 21 and 30 (n= 5 per day). The cytoplasm of interrenal cells located beneath the adrenal capsule (sub-capsular zone, SCZ) were stained lighter than that of interrenal cells located inside the adrenal gland (inner zone, IZ) and contained several vacuoles for each sampling day. Additionally, unlike IZ cells, SCZ cells contained nuclei that were various shapes and surrounded by irregularly arranged membranes, lipid droplets which were not surrounded by a membrane, mitochondria with mostly shelf-like cristae. The arrangement of SCZ cells appears similar to that of zona glomerulosa and also the arrangement of IZ cells to that of zona fasciculata of mammalian adrenal cortex, suggesting the significant signs of zonation in goose adrenal cortex.     

Co-localization of vasoactive intestinal polypeptide and neuropeptide Y immunoreactivities in the nerve fibers of the rat adrenal gland

The co-localization of Vasoactive Intestinal Polypeptide (VIP) with Neuropeptide Y (NPY) or its C-flanking peptide (C-PON) was investigated with immunocytochemistry methods in the adrenal gland of the rat. Most of the VIP immunoreactive (+) nerve fibers found in the capsule/glomerular zone also exhibited NPY or C-PON immunoreactivity (IR). We found that at least two populations of VIP varicose nerve fibers can be observed, the most prevalent exhibited both VIP/NPY or VIP/C-PON IR and the other which was rather scarce lacked NPY or C-PON IR. In the superficial cortex VIP/NPY or VIP/C-PON IR nerve fibers were often associated with capsular or subcapsular vascularization and extended into the zona glomerulosa. In the deeper layers of the adrenal cortex radial fibers were closely associated with the inner vascularization of the zona fasciculata and reticularis. In the adrenal medulla NPY or C-PON immunoreactivity was associated with ganglion neurons as well as chromaffin cells; these last cells were always VIP (-). VIP and NPY/C-PON IR could be co-localized in catecholaminergic nerve terminals of the adrenal cortex but not in the adrenal medulla.     

Structure and dynamics of adrenal mitochondria following stimulation with corticotropin releasing hormone.

The mitochondria of rat adrenals were investigated qualitatively and quantitatively in different functional states of the adrenal cortex. Following stimulation of the animals with corticotropin releasing hormone (CRH), the corticosterone serum levels reached a maximum 1 hour after stimulation with CRH. The amount of inner mitochondrial membrane within the zona fasciculata increased showing a biphasic time course, with a first maximum 2 hours and a second maximum 8 hours after stimulation. In contrast, a significant rise of mitochondrial volume occurred only 24 hours after CRH stimulation. Therefore, the dense vesicularization of mitochondrial cristae may constitute an early process to enhance the steroidogenic capacity of these cells. Within cells of the transition zone between zona glomerulosa and zona fasciculata, we could depict a special type of mitochondria with characteristic crescent-like cristae only seen after stimulation with CRH. This type of mitochondria may represent an intermediate form between mitochondria of zona glomerulosa and zona fasciculata underlining the impressive transformational capacity of adrenocortical mitochondria. After hypophysectomy, zona fasciculata cells contained mitochondria with tubular inner membranes, representing a hypofunctional state. In contrast, the hypofunctional state after hypophysectomy and the hyperfunctional state after stimulation of the adrenal cortex via CRH injection did not appear to correlate with the morphology of mitochondria from the zona reticularis and adrenal medulla.     

大鼠肾上腺的NADPH,NPY,CGRP,SP,c—fos细胞化学特性

目的 探讨肾上腺内分泌组织和神经组织的双重组织学特性。方法 组织化学和免疫组织化学技术,在光学显微下观察ＮＡＤＰＨ、ＮＰＹ、ＣＧＲＰ、ＳＰ、ｃ－ｆｏｓ在大鼠肾上腺的分布。结果 肾上腺皮质分布有ＮＰＹ阳性神经细胞和神经纤维、ＣＧＲＰ阳性神经纤维;肾上腺髓质分布有ＮＡＤＰＨ－ｄ阳性神经细胞和神经纤维、ＣＧＲＰ阳性神经纤维、ＳＰ阳性神经纤维、ｃ－ｆｏｓ阳性神经细胞和神经纤维。肾上腺皮质球状带、网状带、束状带细胞均ＮＡＤＰＨ－ｄ阳性,髓质部分嗜铬细胞ＮＡＤＰＨ－ｄ阳性,部分嗜铬细胞ＮＰＹ阳性,部分嗜铬细胞ＣＧＲＰ阳性,部分嗜铬细胞ＳＰ阳性。结论 大鼠肾上腺接受广泛的非经典递质的神经支配,特别是肽能神经支配的肾上腺实质细胞及髓质嗜铬细胞,能分泌多种神经肽物质。提示肾上腺的内分泌活动不仅受到复杂的神经调节而且也受到自身的活     

ULTRASTRUCTURAL ZONATION OF THE HUMAN ADRENAL CORTEX

Electron microscopic observations of sections from adrenal cortex obtained from a twenty-four-year-old male with hypertension were made. In the zona glomerulosa the cells showed a large nuclear-cytoplasmic ratio. The mitochondria had a dark matrix and plate-like inner structure. The agranular endoplasmic reticulum revealed a close association with lipid granules. The Golgi apparatus was prominent and frequently accompanied by centrioles. Free ribosomes were abundant. The zona fasciculata can be subdivided into two layers. In the outer layer the cells are characterized by numerous large lipid granules. Two kinds of mitochondria are discernible; one with a pale matrix and tubulo-vesicular or vesicular inner structure and the other with a dark matrix and tubulo-vesicular or lamelllform cristae. In the inner zona fasciculata and reticularis there are two types of cells besides the dark cells. The type 1 cells have a compact cytoplasm, elliptic or irregularly shaped mitochondria with a dark matrix and tubulo-vesicular or vesicular internal structure. The type 2 cells are large and ovoid, having a clear transparent cytoplasm. The mitochondria are numerous having a clear matrix and almost exclusively vesicular inner elements. Abundance in agranular endoplasmic reticulum, decreased lipid granules, and cytoplasmic incisions are common in both the inner fasciculata and reticularis. In addition the zona reticularis is characterized by increased number of dark cells, numerous pigment bodies, and the appearance of huge mitochondria.
On the basis of these observation were discussed the significance of the organelles in steroid biosynthesis, mode of hormone secretion, and cyto-genesls in the adrenal cortex.     

Functional and morphologic studies of the adrenal cortex and kidney in ovine toxaemia of pregnancy

Functional and morphologic studies of the adrenal cortex and kidney have been carried out in pregnant sheep with spontaneous or dietary restriction-induced ovine toxaemia. It was found that proteinuria was an inconstant feature and no animal showed glomerular lesions analogous to those found in human preeclampsia; thus ovine toxaemia cannot be regarded as a precise experimental model for human toxaemia of pregnancy. The elevation of blood cortisol levels and the morphologic appearance of the adrenal zona fasciculata found in such animals suggest an adrenal response comparable to that caused by adrenocorticotrophic hormone. In addition, animals with severe disease showed evidence of stimulation of the renin-angiotensin-aldosterone system as reflected by elevated blood renin and aldosterone concentrations and raised renal juxtaglomerular indices. Ultrastructural changes in the adrenal zona glomerulosa and renal juxtaglomerular myoepithelioid cells in toxaemic animals resembled those described in non-pregnant sodium-depleted sheep. The finding of juxtaglomerular peripolar cell mitoses and granule exocytosis, the latter only being previously observed in sodium depleted sheep, together with the ultrastructural changes in the adrenal zona glomerulosa and juxtaglomerular myoepithelioid cells, suggest that sodium depletion may play a role in this disease.     

人胎儿肾上腺的组织发生

用光镜、透射电镜观察了４例胚、９０例胎，４例新生儿肾上腺。结果表明，７周胚已可分辨胎儿皮质和永久皮质，１５周时永久皮质开始分化，出生时初级球状带和束状带已形成，网状带未出现；７周时神经嵴细胞开始迁入肾上腺，１３周时髓质形成，嗜铬细胞出现，１６周时交感神经节细胞出现，出生时仍有少量神经嵴细胞向肾上腺内迁移。     

The zona glomerulosa of the adrenal gland in rats with portacaval shunt. An ultrastructural and morphometric study

The zona glomerulosa of the adrenal glands was studied histologically, ultrastructurally and morphometrically in portacaval shunt (PCS)-bearing rats 4 weeks after surgery. Compared with controls, the zona glomerulosa of rats with PCS showed an increase in thickness and a reduction in the number of lipid droplets in the cells of its inner portion, adjacent to the intermediate zone. Moreover, electron microscopy and morphometry of the cells of the inner portion revealed that qualitative and quantitative changes occur, consisting in a decrease in lipid droplets, an increase in the amounts of smooth endoplasmic reticulum and mitochondria, enlargement of the Golgi apparatus, and the appearance of numerous dense bodies at the vascular cell poles. All the above findings indicate that PCS induces an enhanced activity of the fully differentiated cells of the zona glomerulosa, which may be considered as an adaptive response--mediated by an activation of the renin-angiotensin system--to the lowering of the systemic pressure taking place in this experimental condition.