Morphofunctional features of the secretory cells of the adenohypophysis and adrenal glands of the rat during aging

Karyometric and electron microscopic investigation has been performed in the adenohypophyseal secretory cells, in the cortex and medulla of the adrenal glands in mature (6 months) and ageing (25-26 months) white male rats. The diameter of the corticotropic cell nuclei of the adenohypophysis significantly increases with age. In the secretory cells of the adrenal cortex and medulla no important changes in their parameters are revealed. With age destructive changes of the ultrastructures are manifested variously in different types of the secretory cells in the endocrinic glands studied. In the corticotropic, somatotropic, thyreotropic cells of the adenohypophysis, in spongiocytes of the adrenal cortex, compensatory ultrastructural rearrangements develop; they are aimed to preserve functional activity of the cells mentioned. No age changes are revealed in ultrastructure of chromaffin cells of the adrenal medulla.     

Sex differences in adrenocortical structure and function

Summary Adrenal glands of adult male hamsters are larger and secrete more cortisol than those of females. Stereology was therefore used to study zonal and cellular aspects of development of the adrenal cortex of male and female hamsters. Adrenal glands were studied at weekly intervals from day 21 to day 77 of postnatal ontogenesis. Within this period, body weight did not differ significantly between the sexes. During development, absolute and relative adrenal weights were higher in males; their zona glomerulosa (ZG), zona fasciculata (ZF) and zona reticularis (ZR) become markedly larger than those in females. No marked changes in the volume of individual ZG cells occurred although ZF cells and ZR cells become larger in male than female animals. The total number of adrenocortical cells increased within the period studied, a greater increase being observed in ZG and ZF in males. No distinct sex difference was observed in the number of ZR cells throughout development. From day 56 of postnatal life the adrenal cortex of male hamster contained more parenchymal cells than the female gland. These results thus indicate that sex differences in hamster adrenal cortex depend upon changes in number and size of parenchymal cells.Supported in part by a grant from the Committee of Zoology, Polish Academy of Sciences     

Adaptation of the bank vole (Clethrionomys glareolus Schreber) to conditions of biogeochemical province with abnormally high content of nickel, cobalt and chromium

Morphophysiological characteristics and peculiarities of adrenal gland of bank vole (Clethrionomys glareolus) were studied in the area of natural biogeochemical province with abnormally high content of nickel, cobalt and chromium. The control population inhabited area with usual content of these elements. We used 4-factor analysis of variance to estimate the influence of geochemical conditions, phase of population cycle, sex and reproductive state on the morphophysiological characteristics of animals and functional activity of adrenal gland. Animals from area with high concentration of Ni, Co and Cr show an increase in relative mass of adrenal glands, fascicular zone of adrenal cortex, size of cells and their nuclei. All these changes can be considered as an evidence of increased secretion of glucocorticoids. It is shown that phase of population cycle influences fatness of animals, size of nuclei, cells and adrenal cortex. Females in comparison with males are characterized with higher indexes of liver and adrenal gland, as well as morphometric indexes of adrenal cortex. The maturation of animals is accompanied with increase in body mass, fatness and relative mass of adrenal glands, the size of cortex zone, nuclei and cells themselves. It is supposed that the effect of "geochemical factor" results in intensification of glucocorticoid secretion of adrenal costex, thus increasing non-specific resistance of animals inhabiting area with high concentration of heavy metals. Such factors as "phase of population cycle", "sex" and "reproductive state", influence mineralocorticoid activity, glucocorticoid and androgenic functions of adrenal cortex. Some factors show synergetic effect.     

Orthosympathetic reaction during paroxysmal asphyxia in the rat]

Acute hypercapnic hypoxia provoked by the ligature of the trachea kills the rat in 3 to 5 minutes. Catecholaminemia is immediately increased by adrenaline release from the adrenal medulla for the largest part. Catecholamine contents of the suprarenal glands is not decreased at the time of death.     

Histochemical studies on the developing adrenal gland of Gallus domesticus

Summary The distribution of adrenaline, noradrenaline, aliesterases and non-specific cholinesterases in the cortical and medullary cells and that of ascorbic acid in the cortex have been studied histochemically in sections of adrenal glands from embryonic, juvenile and adult chicken. Both the catecholamines are secreted by the embryonic medulla from the 11th day of incubation but noradrenaline is the more abundant of the two hormones at all stages and it is secreted by the majority of chromaffin cells. There is a tendency for the adrenaline-secreting cells to predominate in the subcapsular layer of the medulla. Both types of chromaffin cells reveal considerable cholinesterase activity consistently from the second half of incubation period onwards.A high concentration of aliesterases and ascorbic acid are developed and maintained in the cortical cords from the time the cortex begins secretory activity, namely, the 10-day incubation stage. Lower concentrations of cholinesterases are also present in the cells of the cortex. The cords of the peripheral zone of cortex show higher concentrations of both the enzymes and ascorbic acid than those of the central zone.From a thesis submitted to McGill University, Montreal, Canada in 1963 in partial fulfillment of the requirements for the degree of Doctor of Philosophy. The work was done during tenure of a Canadian Commonwealth Scholarship.     

Morphology and distribution of chromaffin cells in the adrenal gland of cordylidae (Reptilia,Sauria): A comparative study

The distribution of the adrenaline and noradrenaline chromaffin cells in the adrenal glands of 10 members of the family Cordylidae have been examined. In the genus Gerrhosaurus, all the catecholamine cells lie on the surface of the adrenal gland, forming a continuous envelope of one or two layers of cells that mainly contain noradrenaline (NA). In the genus Platysaurus, the chromaffin envelope is intermittent. There are relatively large tracts of interspersed interrenal tissue containing some adrenaline cells (A). Islets of chromaffin cells are scattered between these interrenal tracts. In the genus Pseudocordylus and the genus Cordylus, the superficial chromaffin cells tend to gather into a multilayered dorsal mass, containing mainly NA cells. Inside the interrenal parenchyma, there are always numerous chromaffin islets, containing mainly A cells.     

Ultrastructural pathology of the adrenal gland in Cushing's syndrome

The ultrastructural pathology of the adrenal glands was studied in fifteen cases of Cushing's syndrome. Some specific features correlated with the pathological aspects of adrenals were found. In the hyperplastic adrenal cortex the cytoplasms contained a rich smooth endoplasmic reticulum and many mitochondria. Increased lipid-pigment complexes were found especially in the compact cells. In adenomas, the clear cells showed large lipid vacuoles; the compact cells presented anisomorphous mitochondria, a well developed smooth endoplasmic reticulum and many pigment bodies. The nuclei of adenomatous cells were irregular, with deep invaginations. In adrenal carcinomas, the pleomorphism of nuclei, nucleoli, mitochondria, and smooth endoplasmic reticulum was more obvious. Absolutely reliable characteristics proving malignancy at ultrastructural level do not, however, exist. The steroidogenic activity of both hyperplastic and tumoral adrenal glands can be assessed using the agranular endoplasmic reticulum and the mitochondria as functional parameters.     

Essential Role for the Lymphostromal Plasma Membrane Ly-6 Superfamily Molecule Thymic Shared Antigen 1 in Development of the Embryonic Adrenal Gland

Thymic shared antigen 1 (TSA-1) is a plasma membrane protein of the Ly-6 superfamily expressed on thymocytes, thymic stromal cells, and other cells of the hematopoietic system. TSA-1 is also expressed in other nonhematopoietic tissues, in particular, embryonic and adult adrenal glands. To address the function of TSA-1, we generated mutant mice in which TSA-1 expression was inactivated by gene targeting. Here we show that deletion of both TSA-1 alleles results in abnormal adrenal gland development and midgestational lethality due to cardiac abnormalities. We also report that TSA-1-deficient adrenal glands have significantly reduced levels of the catecholamines noradrenaline and adrenaline. We conclude that TSA-1 is required for normal embryonic development but that deletion of its expression does not obviously impair lymphoid development.     

A histochemical analysis of adrenal lipids in Ox and Sheep

Summary The adrenals of Ox and Sheep were analysed by various histochemical methods available for lipids. The cortex was found to be rich in choline containing phospholipids, unsaturated phospholipids and masked lipids compared with the medulla. Unsaturated lipids and free fatty acids were more abundant in the cortex. There was slightly more cholesterol and plasmalogen in the medulla than the cortex. Formaldehyde fixation apparently increased the relative number of carboxyl groups in the adrenaline storing cells. Osmium tetroxide reacted with catechol amines and differentiated between the adrenaline and noradrenaline storing cells in the ox medulla. The histochemical results are compared with previous biochemical findings on the nature and distribution of adrenal lipids.     

A correlated stereological and functional studies on the long-term effects of ACTH on rat adrenal cortex

The aim of the study was to investigate the effect of prolonged ACTH administration on quantitative structural changes of the rat adrenal cortex and on function of its cells with particular emphasis on correlation of the results of biochemical estimations with stereologic parameters. Daily injections of 20 micrograms ACTH (Synacthen, Depot) for 35 days results in a marked enlargement of the cortex due to an increase in the volume of all the zones. This increase depends upon hypertrophy and hyperplasia of parenchymal cells. At day 21 of experiment the number of parenchymal cells markedly decreased if compared with day 14, the lost of cells being observed mainly in the zona reticularis. Prolonged ACTH treatment only insignificantly changed serum corticosterone concentration and--if calculated per mg of adrenal weight--did not change adrenal corticosterone concentration and 11 beta-hydroxylase activity and decreased corticosterone output by adrenal homogenate. If expressed per adrenocortical cell or per pair of glands, ACTH increased corticosterone concentration and 11 beta-hydroxylase activity while the drop in corticosterone output occurred only at days 28 and 35 of experiment. The striking differences in and among various functional parameters depicting adrenal steroidogenesis show for necessity--in case of long-term experiments leading to hypertrophy or atrophy of the gland--of using coupled stereologic and biochemical techniques which better evaluate the cytophysiological state of adrenocortical cells.     

On the presence of chromaffin cells in the adrenal cortex: their possible role in adrenocortical function

Using light and electron microscopy, we have observed the presence of rays containing medullary tissue extending across the cortex of rat adrenal glands. Within these rays chromaffin cells, as well as collagen and nerve fibers, were present. It is suggested that these endocrine cells may have a paracrine function within the cortex, possibly via their secretory product.     

Morphometric and histological studies on the adrenal glands of the camel Camelus dromedarius

The adrenal gland of the camel consists of an outer cortex and an inner medulla. The general disposition of the cortex and medulla, however, differs occasionally from that of other mammals. Extensions of medulla could reach as far as the periphery of the cortex. Islet of medullary tissue may be found in sections of the cortex and cortical tissue consisting of all zones of the cortex may occur around arteries or nerves in the medulla. The medulla may be separated from the cortex by connective tissue especially in old camels. The arrangement of noradrenaline-secreting cells is different from that in other ruminants; they are found in groups scattered between the adrenaline-secreting cells. Bundles of smooth muscle occur in venules at the corticomedullary interface. Accessory adrenal glands are found embedded in the renal fat. They are similar in structure to the adrenal gland. The adrenal cortex forms 74% of the volume of the gland and the ratio of the cortex to medulla is 4:1. The zona glomerulosa, fasciculata and reticularis constitute about 13%, 53%, and 29% by volume of the cortex, respectively.     

Microwaves in the Induction of Immune Response to VI-Antigen

In this work the possibility of using microwaves (MW) for immuno-modulation in the immunization of animals with thymus-independent antigen was studied. The projection zones of the thyroid and adrenal glands of the test animals were subjected to the action of decimeter MW, while the corresponding zones of control animals were subjected to imitation MW. The endocrine activity of rabbits was estimated by radioim-mune methods. Vi-antigen was shown to be a thymus-independent antigen for rabbits, according to the results of fluorescent probes to study the structural rearrangements in surfaces of thymocyte membranes and their nuclei, which reflect early changes during the physiological activation of cells. The irradiation by MW on the projection zone of the thyroid was accompanied by a decrease in the glucocorticoid activity of the adrenal cortex and a simultaneous pronounced immunostimulating effect. MW irradiation of the zone of the adrenal glands was accompanied by immunosuppression in combination with enhanced glucocorticoid activity of the adrenal cortex.     

Sex differences in adrenocortical structure and function

Summary Adrenal glands of adult female rats are heavier than the glands of corresponding male rats. Postpubertal orchiectomy increases the adrenal weight, an effect restored by testosterone replacement. Under the same conditions ovariectomy of 8 weeks duration does not change the adrenal weight while estradiol replacement enhances the relative adrenal weight.Karyometric studies have shown that nuclei in the female zona fasciculata cells are larger (app. 18%) than those of the male. Similar but only slight differences (2%) were observed in the zona reticularis. Orchiectomy results in enlargement of cell nuclei within all zones of the adrenal cortex; testosterone replacement has the opposite effect. Ovariectomy of 8 weeks duration slightly enhances the volume of nuclei of the zona glomerulosa cells, has no effect on the nuclei of the zona fasciculata and reduces the volume of nuclei in the zona reticularis. Estradiol replacement reduces the volume of nuclei of the zona glomerulosa cells but increases nuclear volume in the zona fasciculata and in the zona reticularis.Thus testosterone has an inhibitory effect on the adrenal cortex of the rat while the physiologic effect of estradiol on the morphology of this gland, particularly on the zona fasciculata cells is rather dubious.The author wishes to thank Mrs. B. Westerska and Miss K. Siejak for excellent technical assistance.This research was supported in part by a grant from the Zoological Committee, 2nd Department, Polish Academy of Sciences.This paper is dedicated to the memory of late Kazimierz Mietkiewski, M.D., Ph.D. whose encouragements and suggestions were most stimulating for my work.     

Participation of adrenal hormones in the genesis of arterial hypertension of hypothalamic origin]

Continuous many-hour stimulation of the hypothalamic negative emotiogenic centres (ventromedial nuclei) evoked a stable arterial hypertension with a peculiar phasic dynamics of the adrenal secretory activity in waking immobilized rabbits. Bilateral extirpation of the adrenal glands decreased the initial level of the average arterial pressure, inhibiting development of stable arterial hypertension. Many-hour stimulation of the mentioned structures also produced stable arterial hypertension in adrenalectomized rabbits if preceded by the administration of hydrocortisone together with adrenaline. Against the background of separate administration of hydrocortisone and adrenaline to adrenalectomized rabbits, many-hour stimulation of the ventromedial hypothalamus evoked a short-term rise in the arterial pressure. A conclusion was drawn that activation of hormones of the adrenal cortical and medullary layers played an important part in the formation of stable arterial hypertension in rabbits under continuous many-hour stimulation of the hypothalamic negative emotiogenic centres.     

The effects of chronic SRIH-14 and octreotide administration on the pituitary-adrenal axis in adult male rats

The effects of chronic treatments with SRIH-14 and octreotide on pituitary corticotropes (ACTH cells) and on the adrenal cortex of male Wistar rats were examined. Adult males received two daily s.c. injections of 20 microg/100 g of body weight of either SRIH-14 or octreotide for 28 consecutive days. ACTH cells were studied using a peroxidase-antiperoxidase immunocytochemical procedure. Morpho-metry was used to evaluate the changes in cell and nuclear volumes (microm3) and volume densities (%) of ACTH-immunoreactive cells. The adrenal cortex was analyzed by histological and morphometric methods. A significant (p<0.05) decrease in body weight and in the absolute weights of the pituitary and adrenal glands was observed in both treated groups. Morphometric parameters of ACTH cells in both treated groups were not significantly (p>0.05) different than in control rats. The absolute volumes of the adrenal gland and adrenal cortex were significantly (p<0.05) decreased in both treated groups. The absolute and relative volumes of the zona glomerulosa (ZG), as well as the cellular and nuclear volumes of the ZG were significantly (p<0.05) decreased in the both treated groups. In rats treated with SRIH-14 and octreotide, the absolute and relative volumes of the zona fasciculata (ZF) and zona reticularis (ZR), as well as their stereological parameters, did not change significantly (p>0.05). The aldosterone levels in the SRIH-14 and ocreotide-treated groups were significantly (p<0.05) decreased - by 13% and 19%, respectively. The concentration of ACTH and corticosterone did not change significantly. Together, these findings show that SRIH-14 and octreotide administration affected the morphological characteristics of the adrenal ZG in a similar manner, and brought about a decrease in plasma aldosterone concentration. These treatments did not affect pituitary ACTH cells or adrenal ZF and ZR functioning.     

Hypertrophy and altered activity of the adrenal cortex in Homer 1 knockout mice

Homer 1 gene products are involved in synaptic transmission and plasticity, and hence, distinct behavioral abnormalities, including anxiety- and depression-like behaviors, have been observed in Homer 1 knockout (KO) mice. Here we report that Homer 1 KO mice additionally exhibit a pronounced endocrine phenotype, displaying a profoundly increased adrenal gland weight and increased adrenal/body weight ratio. Histological examinations of Homer 1 deficient adrenal glands revealed an increased size of the adrenal cortex, especially the sizes of the zona fasciculata and zona glomerulosa. Moreover, the plasma corticosterone and aldosterone were higher in Homer 1 KO than wild-type (WT) mice while the plasma ACTH levels were not different between the genotypes. The in vivo ACTH test revealed that corticosterone and aldosterone plasma levels were higher in saline injected Homer 1 KO mice than in WT mice (saline injected mice served as controls for the respective groups of ACTH-injected animals), but the magnitude of steroid responses to ACTH was similar in both genotypes. In contrast, an in vitro experiment performed on isolated cells of adrenal cortex clearly showed increased production of both steroids in response to ACTH in Homer 1 KO cells, which is in line with an ~8-fold increase in the expression of ACTH receptor mRNA in the adrenal cortex of these mutants. These results, together with the detection of Homer 1 mRNA and protein in the adrenal cortex of WT mice, indicate that Homer 1 directly affects the steroidogenic function of the adrenal glands.     

On the uptake of exogenous catecholamines by adrenal chromaffin cells and nerve endings

Summary Light-microscopic autoradiography has revealed characteristic labelling patterns in adrenal medullary cells following the intravenous administration of different catecholamines. The uptake patterns for [³H] dopa, [³H] dopamine, [³H] noradrenaline and [³H] adrenaline have been compared. In all cases A cells were more active than NA cells and cells situated in the zone nearest the cortex demonstrated a markedly higher rate of uptake than central cells. It was concluded that adjacent chromaffin cells with very similar morphology may differ as much as 50 fold in their capacities to incorporate exogenous amines. The adrenergic nature of the innervation of the vessels of the adrenal cortex and capsule in the mouse was confirmed.     

Effects of tropic hormones on ultrastructure and synthesis of androgens in adrenals of male pseudohermaphrodite rats

Summary Ultrastructural and biochemical study of the adrenals in the pseudohermaphrodite (tfm) rat reveals hypertrophic adrenocortical cells. The cytoplasm of the cells in the zonae fasciculata and reticularis contains an exceptionally well developed smooth endoplasmic reticulum closely applied to mitochondria and lipid droplets. Mitochondria are more numerous than in normals and have especially abundant tubular cristae. More lipid droplets (appearing as empty vacuoles) are surrounded by pleomorphic mitochondria.The incubation study indicates that the capacity of rat adrenal cortex of producing androgens is greater in tfm than in normal animals. Hypophysectomy and castration result in a significant decrease in androgen biosynthesis by tfm rat adrenals and cause a reduced concentration of plasma testosterone. Administration of tropic hormones to hypophysectomizedcastrated rats appears to stimulate their adrenal androgenesis. It is suggested that in tfm rats the higher than normal luteinizing hormone (LH) together with adrenocorticotropic hormone (ACTH) stimulates the hypertrophy of cellular organelles in the adrenal cortex and causes an accompanying increase in androgenic steroids which may be responsible, at least in part, for the increased level of plasma androgens.     

The determination of the adrenal medullary cell fate during embryogenesis

One subset of neural crest cells, the sympathoadrenal precursors, undergoes a switch in phenotype expression, when they invade the adrenal anlagen and become associated with adrenocortical cells. To investigate the mechanisms responsible for the conversion of noradrenaline synthesizing precursors to adrenaline producing endocrine chromaffin cells we studied the role of glucocorticoids on the initial induction of adrenaline synthesis in embryonic adrenals and cultures of highly purified chromaffin precursor cells. We could show that in vivo differentiation of rat chromaffin precursors commences between 16.3 and 17.3 days of gestation. While adrenaline and the activity of the enzyme phenylethanolamine N-methyltransferase (PNMT), which converts noradrenaline to adrenaline, were present at Embryonic Day 17.3 (E17.3), they were not detectable in E16.3 adrenals. Small amounts of corticosterone were present in E16.3 adrenals and plasma, but in parallel with the initial induction of adrenaline biosynthesis, a sharp rise in organ and plasma glucocorticoid levels occurred until E17.3. Chromaffin precursor cells, isolated at E16.3 and cultured for 4 days, failed to express PNMT activity and adrenaline. However, 0.1 nM dexamethasone was already sufficient for the initial induction of adrenaline and its synthesizing enzyme. Specific glucocorticoid binding of freshly isolated chromaffin (precursor) cells revealed a developmental increase during embryogenesis, yet no glucocorticoid binding sites were detectable in chromaffin precursor cells at E16.3. They appeared at E17.3 in parallel with the initial induction of adrenaline biosynthesis and the enormous rise of adrenal and plasma corticosterone levels. We therefore conclude that glucocorticoids are essential and sufficient to trigger the differentiation of noradrenergic sympathoadrenal precursors to adrenergic chromaffin cells after a functional glucocorticoid receptor system has been established.