Ret protein expression in adrenal medullary hyperplasia and pheochromocytoma

Ret is a developmentally regulated tyrosine kinase involved in formation and maintenance of the nervous system. Ret mutations predisposing to pheochromocytomas and medullary thyroid carcinomas occur in multiple endocrine neoplasia (MEN) syndromes 2A and 2B. Biochemical studies have demonstrated overexpression of Ret mRNA and protein in pheochromocytomas compared to normal adrenal medulla. However, the cellular distribution of Ret in the normal human adrenal and in hyperplastic lesions that antecede pheochromocytomas are unclear. The present investigation was undertaken to resolve the histological distribution of Ret in the normal human adrenal, in pheochromocytomas evolving from adrenal medullary hyperplasia in MEN2A and in sporadic pheochromocytomas. Ret expression was studied by immunohistochemistry using both a polyclonal and a monoclonal antibody, with confirmation by immunoblotting of representative cases. Only occasional cells stained for Ret in the normal adrenal, consistent with the distribution in adult adrenals of other species. Heterogeneous, progressively increased Ret expression was observed during the evolution of pheochromocytomas. In both normal and neoplastic adrenal, the most intense immunoreactivity was observed in cells with neuron-like features. Our finding that Ret is not expressed at high levels in the early stages of disease suggests that elucidation of mechanisms that regulate Ret expression is required for understanding the pathobiology of MEN2A. The association of high-level Ret expression with neuronal morphology suggests that the variable overexpression of Ret in pheochromocytomas might in part be an epiphenomenon, reflecting the known phenotypic plasticity of these tumors.     

Innervation of human adrenal gland and adrenal cortical lesions

The innervation of the human adrenal gland and of cortical lesions was studied in sections of cortical tissue (n=10), hyperplastic cortical tissue (n=3), and tissue from cortical adenomas (n=5) and carcinomas (n=6). The presence and distribution of nerve structures containing neuronal markers indicating sympathetic and parasympathetic innervation were studied by immunohistochemistry and the co-existence and co-localization patterns of the different markers by immunofluorescence. The cortex and hyperplastic cortical tissue had a moderate to rich supply of nerve structures containing the typical neuronal markers: protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), small vesicle synaptic protein type 2 (SV2), and nerves showing immunoreactivity to the adrenergic marker tyrosine hydroxylase (TH). All these immunoreactive nerves were located predominantly adjacent to blood vessels, but also among parenchymal cells. The cortex showed numerous nerve structures containing the neuropeptide substance P (SP), neuropeptide Y (NPY) and vasoactive intestinal protein (VIP), but few nerves containing these peptides were seen in hyperplastic cortical tissue. Typical markers were occasionally observed in cortical adenomas but were not found in carcinomas, except in a few cases where PGP 9.5 and NSE were present, but only adjacent to necrotic areas. Nerves containing NPY and VIP occurred in varying numbers in both adenomas and carcinomas. NPY- and VIP-immunoreactive nerve structures were seen mostly alongside blood vessels. There were several types of co-existence. For instance, NSE/VIP-, TH/VIP- and TH/NPY-immunoreactive nerve structures were often seen in the same trunk, but were only partly co-localized. Received: 19 January 1999 / Accepted: 18 May 1999     

Lack of effect of opioid compounds on angiotensin II responses of bovine adrenal medullary cells

Angiotensin II (10 nM) increased basal adrenaline and noradrenaline secretion from cultured bovine adrenal chromaffin cells by 2.5- to 3-fold and 4- to 6-fold, respectively, and stimulated basal accumulation of inositol phosphates more than 2-fold. Etorphine and diprenorphine in the range 10⁻⁹ to 10⁻⁵ M had no effect on the catecholamine secretion induced by angiotensin II, and, at 10⁻⁸ and 10⁻⁵ M, had no effect on angiotensin II-induced inositol phosphate accumulation. The functions of adrenal medullary opioid receptors remain to be determined.     

大鼠双侧肾切除后肾上腺皮质球状带细胞苯二氮受体的量变

用选择性作用于外周型苯二氮(艹卓)受体的标记配体[~3H]PK11195检测了Wistar大鼠分离的肾上腺皮质球状带细胞上苯二氮(艹卓)受体的结合动力学参数,结果表明Wistar大鼠在双侧肾切除后36小时、其球状带细胞苯二氮(艹卓)受体结合[~3H]PK11195的最大容量(B_(max))显著高于假手术组和正常对照组的相应数值,三组的B_(max)分别为12.1±1.1、6.6±1.3和5.6±1.8(pmol/10~6cells),三组的K_D值则无显著差异。     

Fine structure of adrenal medullary grafts in the pain modulatory regions of the rat periaqueductal gray

Summary Recent findings in our laboratory indicate that adrenal medullary grafts produce significant alterations in pain sensitivity. Electron microscopic studies were undertaken to correlate these behavioral changes with the neural interactions of the host and graft tissue in the periaqueductal gray. A striking change found 8 weeks after transplantation is that pronounced myelination has taken place both in the graft and in the host tissue. The new myelin formation in the graft has the typical appearance of PNS myelination and, in the host the appearance of CNS myelination. The endothelial cells of the capillaries in the grafted tissue are attenuated and fenestrated in contrast to those of the surrounding parenchymal tissue of the host. By 8 weeks, the graft becomes heavily encapsulated with collagen, while the host CNS tissue develops layers of glial processes outlining the graft. However, collagen and glial layers apparently do not form an absolute barrier to either cellular or humoral interaction between the host and graft tissue. Chromaffin cells can be found protruding into the host CNS tissue and sometimes forming synapses with presumably the host neuronal processes. Grafted chromaffin cells may participate as both postsynaptic and, less often, as presynaptic components of synaptic junctions. The behavioral relevance of these synaptic contacts is unclear, since similar implants of adrenal medullary tissue into the dorsal spinal cord subarachnoid space, which also induce potent analgesia, do not contain synapses. Thus, it is more likely that behavioral changes are brought about by diffusion of neuroactive substances from grafted chromaffin cells to host receptors.     

Interaction between dendritic cells and nerve fibres in lymphoid organs after oral scrapie exposure

In transmissible spongiform encephalopathies (TSEs), the infectious agent, called PrPsc, an abnormal isoform of the cellular prion protein, accumulates and replicates in lymphoid organs before affecting the nervous system. To clarify the cellular requirements for the neuroinvasion of the scrapie agent from the lymphoid organs to the central nervous system, we have studied, by confocal microscopy, the innervations within Peyer’s patches, mesenteric lymph nodes and the spleen of mice in physiological conditions and after oral exposure to prion. Contacts between nerve fibres and PrPsc-associated cells, dendritic cells (DCs) and follicular dendritic cells (FDCs), were evaluated in preclinical prion-infected mice. Using a double immunolabelling strategy, we demonstrated the lack of innervation of PrPsc-accumulating cells (FDCs). Contacts between nerve fibers and PrPsc-propagating cells (DCs) were detected in T-cell zones and cell-trafficking areas. This supports, for the first time, the possible implication of dendritic cells in the prion neuroinvasion process.     

Reinvestigation of the effect of orexin A on catecholamine release from adrenal chromaffin cells

Orexins have been shown to be implicated in the regulation of adrenal medulla functions. However, there are still inconsistent investigations on the effects of orexins on catecholamine release from chromaffin cells in varying species. In the present study, using the carbon-fiber amperometry, we investigated whether orexin A would stimulate catecholamine release from rat and mouse adrenal chromffin cells. Puff application of orexin A dose-dependently induced amperometric currents in the cultured rat chromaffin cells, which was completely blocked by the selective OX1R antagonist SB-334867 or by the removal of extracellular calcium. Likewise, in the mouse adrenal medulla slices, orexin A also induced catecholamine release mainly through the activation of OX1R. These results gain insight into our understanding of the pharmacological relevance of orexin system in modulating neuroendocrine functions.     

Characterization of catecholamine release from deer adrenal medullary chromaffin cells

Isolated adrenal medullary chromaffin cells maintained in culture have been widely used to study neurosecretory events. Many of these studies have been conducted using cells obtained from the bovine adrenal. In this study we have cultured chromaffin cells from an alternative large animal model, the deer, and have conducted the first characterization of secretion from this preparation. Cervine chromaffin cells, preloaded with [3H]noradrenalin, displayed a strong secretory response to the cholinergic agonist carbachol, with a maximal secretion of approximately 28% cell content over 15 min. This response was reproduced by nicotinic but not muscarinic agonists and was similarly inhibited by nicotinic but not muscarinic antagonists. Nicotine-evoked secretion measured over a 15 min time period was inhibited approximately 50% by the L-type Ca2+-channel antagonist nifedipine and approximately 20% by N-type (omega-conotoxin GVIA) or N, P/Q-type (omega-conotoxin MVIIC) antagonists. In contrast the response was unaffected by omega-agatoxin IVA, a P/Q-type antagonist. In addition to nicotinic receptor stimulation, activation of PACAP or histamine H1 receptors resulted in a concentration-dependent increase in secretion. PACAP was approximately two-fold more effective than histamine although both were weaker secretagogues than nicotine. In contrast, cervine chromaffin cells did not respond to angiotensin II or bradykinin, two agents known to stimulate secretion from bovine chromaffin cells. These data provide an initial characterization of the secretory response from cervine adrenal medullary chromaffin cells indicating that there are marked similarities but also potentially significant differences between them and their far more extensively described bovine counterparts.     

Weight and shape of the human adrenal medulla in various age groups

Summary The weight and shape of the adrenal medulla were studied in 118 adrenal glands obtained at autopsies of 62 patients (38 males and 24 females) between 0 and 52 years of age. In adolescents 15 years and older and adults cases of sudden death only were entered in this study. The weight was calculated using morphometric measurements done on serial sections of the glands. In the new-born, the medulla accounts for less than 1% of the total volume of the adrenal gland. Though there is a rapid growth of the adrenal medulla after birth, the percentage of adrenal medullary volume at all age levels during childhood and adolescence is lower than in adults where it constitutes 9% of the total adrenal volume on the average. This corresponds to an average medullary weight of 0.43 g. In the new-born, the medulla consists of a thin plate made up of immature medulloblasts. Within a few months these are transformed into mature medullary cells; the shape of the medulla soon approaches that of the adult gland: an increase in thickness around the central vein and flat processes into the alae. With increasing age the cortico-medullary border becomes irregular and ragged. Especially in the vicinity of the central vein an intermingling of medullary and cortical cell complexes is found. An unequivocal diagnosis of genuine adrenal medullary hyperplasia can be established only by weighing the dissected medulla or by applying morphometric methods.     

Assessment of malignancy in adrenal cortical tumors

The roles of the pathologist in assessing adrenal cortical tumors are, first, to differentiate adenoma from carcinoma, and, second, to assess prognosis when the diagnosis of malignancy is made. How easy is it to achieve these goals with standard histology and immunohistochemistry, and to what extent can molecular analysis help?     

Innervation distribution pattern,nerve ending structure,and fiber types in pigeon skeletal muscle

Four fiber types have been characterized in different pigeon skeletal muscles according to their innervation pattern (nerve ending structure and innervation distribution) and histochemical properties (SDH and m-ATPase activities). All fast fibers, types IIA and IIB, present aggregated distribution of their nerve endings with “en plaque” structures and very low innervation frequencies. The two kinds of slow fibers recognized are multiple innervated and present higher innervation frequencies. However, type I fibers have nerve terminals in small knobs with uniform localization, whereas type III fibers present “en grappe” nerve endings, which tend to be randomly distributed. Fiber type composition of skeletal muscles has been found closely related to their biomechanical function. Fast fibers are predominant in muscles with an active role in locomotive movements, whereas slow fibers are mainly or exclusively located in postural muscles. © 1993 Wiley-Liss Inc.     

γ－干扰素样物质在肾上腺皮质腺肿瘤中的表达

为了检测γ－干扰素（ＩＦＮ－γ）是否在人肾上腺组织和肾上腺皮质腺肿瘤中有表达，我们对包括瘤旁肾上腺组织１４例；肾上腺皮质腺瘤１７例；皮质腺癌７例的组织标本，用抗大鼠ＩＦＮ－γ单克隆抗体ＤＢ－１做为一抗，采用ＡＢＣ法做免疫组化染色。结果：ＩＦＮ－γ样免疫反应阳性物可在１００％（７／７例）肾上腺皮质腺癌，４７％（８／１７例）皮质腺瘤细胞内和３０％腺癌（２／７例）和腺瘤（５／１７例）组织内毛细血管内皮细胞中检出，瘤旁肾上腺组织内未见阳性反应细胞。结论：（１）ＩＦＮ－γ的存在与肾上腺皮质肿瘤的分化有关，分化越低，ＩＦＮ－γ的阳性率越高。（２）肾上腺皮质肿瘤内的毛细血管内皮细胞可能具有ＩＦＮ－γ受体或能产生ＩＦＮ－γ。另外，在肿瘤组织内还发现一些树突状ＩＦＮ－γ阳性细胞，其存在意义尚待探讨。     

Diagnostic and molecular aspects of adrenal cortical tumors

Adrenal cortical diseases are relatively rare but tumors are the most common in diagnostic practice. This is reflected by the content of this review. By studying familial syndromes in which they occur more frequently and the pathways involved in steroidogenesis and cortical growth, the molecular genetics of these tumors is being unraveled. Genome-wide approaches have also been helpful. The emerging data may complement standard histological investigation in diagnosis and prognosis.     

Melatonin and the adrenal cortex: relationship to territorial aggression in mice

The behaviour of male TO mice on confrontation after a period of group housing (3 mice/group) was recorded, both in terms of group and individual aggression measures. A series of treatments was carried out aimed at identifying the relative contributions of corticosteroids (CS) and melatonin (MEL) to control of fighting. Adrenalectomy (ADX) and aminoglutethimide injections (AMG) were used to reduce CS both chronically and acutely, and repeated restraint stress (RES) to increase CS levels chronically. MEL was administered daily (5 days), alone or in combination with other treatments. Control group treatments included sham operation for ADX (SO), and the appropriate saline injections. Untreated groups were also tested. AMG and ADX reduced aggression to a similar extent on all measures. MEL potentiated fighting, but this effect was reversed when given in combination with ADX or AMG. RES was associated with a significant shift towards submission on individual measures, but no significant change in group measures. RES+MEL treatment did not cause any significant changes. SO animals behaved normally, but again, the aggressogenic MEL effect was not seen after SO + MEL. The results are interpreted in terms of partial inhibition of CS output by MEL.     

Human fetal tissues grafted to rodent hosts: structural and functional observations of brain,adrenal and heart tissues in oculo

Summary The potential for growth and development of human tissue grafts was explored by transplantation to the anterior chamber of the eye of rats and mice. Tissues were obtained from therapeutic abortions, performed in the eighth to twelfth week of gestation, using a slight modification of routine vacuum aspirations. Recipients were either adult rats immunosuppressed with cyclosporin A and protected with antibiotics, or nude immunodeficient Balb C mice. Catecholamine-rich tissues such as chromaffin cells from the adrenal medulla, sympathetic ganglia, central dopamine neuroblasts from the substantia nigra, and noradrenaline neuroblasts from the locus coeruleus all survived grafting, and in many cases formed nerve fibers that invaded the host iris. Similarly, central serotonin neurons from developing raphe nuclei grafts were able to innervate host irides. Human fetal cerebellar and cerebral cortical transplants continued their development in rat host eyes. Extracellular recordings from such cerebellar and cortical grafts revealed spontaneously active cells with immature action potential waveforms. Spinal cord grafts also survived and contained substance P-immunoreactive neurons. Dorsal root ganglia were able to form nerve fibers invading the host iris, as evidenced by neurofilament immunohistochemistry. Heart tissue survived and manifested spontaneous rhythmic contractions in oculo. Both human cortex cerebri and heart tissue grafts became innervated by sympathetic adrenergic nerve fibers from the rat host iris. Thus both graft-to-host and host-to-graft neuronal connections may be established between man and rat. Taken together, these data suggest that transplantation of human fetal nervous tissues to the anterior chamber of immunosuppressed or immunodeficient rodent hosts yields a unique model system for studies of human brain development, developmental disturbances, connectivity, and the action of drugs.     

Presence of immunoreactive atrial natriuretic peptide in nerve fibres and conduction cells in the conduction system of the bovine heart

Previous findings of atrial (A-type) natriuretic peptide (ANP) in nervous tissue, such as the brain and the superior cervical ganglia, led us to investigate the possible occurrence of ANP in nervous tissue in the heart. The distribution of ANP in the bovine heart, particularly its conduction system, was examined by the use of immunohistochemical methods and an antiserum against -hANP. ANP immunoreactivity was frequently detected in atrial myocytes and in the Purkinje fibres of the AV-bundle, and was sometimes seen in the Purkinje fibres of the bundle branches and their ramifications. On the other hand, ANP immunoreactivity was never seen in the conduction cells of SA- and AV-nodes. ANP immunoreactivity was also detected in small nerve-fibre varicosities, mainly in the AV-node and AV-bundle. Most of these varicosities were located in the proximity of the conduction cells, but some occurred close to fine blood vessels or in the walls of arterioles. These observations show for the first time that ANP immunoreactivity is present not only in atrial myocytes and conduction cells but also in nerve-fibre varicosities in the conduction system. The observations suggest that ANP may act as a neuromodulator and/or neurotransmitter in the conduction system.     

The nerve distribution in the testis of the cat

The autonomous innervation of the feline testis was investigated by immunohistochemistry and a modified acetylcholinesterase technique. The nerves reach the testis mainly by two routes: (1) with testicular artery and pampiniform plexus to the cranial extremity (funicular contribution), (2) from the epididymal tail to the caudal extremity (caudal contribution). Within the tunica albuginea the funicular contribution supplies the cranial two thirds, whereas the caudal third of the tunica receives its nerves via the ligamentous connection between testis and epididymal tail. The nerve bundles accompanying the testicular artery give branches to the arterial wall and the pampiniform plexus. When reaching the cranial testicular pole the bundles separate; the majority of them pass into the centrally located mediastinum testis, another large portion enters the tunica albuginea, particularly on its epididymal side. The septula testis are innervated from both sides, that is from the mediastinum and from the tunica albuginea. In the cat, contrary to other mammals, all septula are innervated. Furthermore, nerve fibers occur regularly within the testicular lobules. Generally, the testicular nerves of the cat are unmyelinated and mainly vascular nerves, but fibers are also found within the connective tissue compartments of the testis. The vast majority of all autonomous testicular nerves are postjunctional sympathetic fibers. Terminal ramifications of cholinergic fibers are exclusively observed in the wall of medium-sized arterioles within mediastinum, septula and lobuli testis. Neuropeptide Y is the most frequent peptidergic transmitter in feline testicular vascular plexuses. The amount of calcitonin gene-related peptide-positive fibers is also remarkably high in the testis, but prefers a location within the stroma of the tunica albuginea, mediastinum and septula. In the cat, Leydig cells occur not only in intertubular locations, but also as intratunical and mediastinal Leydig cells. In all three localizations solitary nerve fibers are observed between Leydig cell groups. These fibers are generally dopamin-beta-hydroxylase- and tyrosine hydroxylase-positive, some contain calcitonin gene-related peptide and, very few, substance P.