Behavioral and histochemical observation in rotational rat with adrenal medulla transplantation

It is widely known that rotation rat is prepared by destroying substantia nigra pars compacta in the rat. Adrenal medulla was transplanted to the rat. The transplantation was judged to be effective in the rat to which adrenal medullary cell was transplanted as the number of rotation decreased evidently in the rat with transplanted adrenal medulla even in the change in the rotating motion after actual transplantation. On the other hand, many transplanted cells transformed into nerve cell which was considered to have produced dopamine were observed in the rat in which transplantation was judged as effective in the histochemical assessment made of the group undergone transplantation of the adrenal medulla. However, in the rat in which no decrease in the number of rotation was observed, only the transplanted cells which were presumed to be the primary adrenal medullary cells were existent. The fact suggests that take of dopamine-producing cells is necessary to see decrease in rotating motion which is found to be the judging criterion for clinical effect.     

Autologous transplantation of adrenal medulla in Parkinson's disease. 18-month results

Eighteen of 19 patients who underwent autologous adrenal medullary transplantation to the right caudate nucleus have been followed up for 18 months. During the course of this study, a statistically significant improvement was noted in percent "on" time, percent "on" time without dyskinesia, activity of daily living (ADL) scores during the "on" stages, and ADL, motor, and Schwab-England scores during the "off" stages. Benefits tended to be maximal at 6 months and to gradually lessen thereafter, although statistically significant improvement in comparison with baseline was still present at 18 months for ADL, motor, and Hoehn-Yahr scores during the "off" stages. Almost all parameters had deteriorated by 18 months compared with 12 months, including those remaining significantly improved in comparison with baseline. These patterns were similar for each of the three participating centers. Complications were largely restricted to the perioperative period.     

肾移植后西罗莫司致不良反应的文献分析

背景：西罗莫司是一新型免疫抑制剂,肾毒性小,已在中国用于防治肾移植后排斥反应。 目的：分析肾移植后应用免疫抑制剂西罗莫司所致不良反应的临床特征和相关因素。 方法：计算机检索2002/2009中国知网数据库、万方数据资源的中国期刊全文数据库国内文献收载的西罗莫司不良反应报道资料,检索词“西罗莫司、肾移植”,采用文献计量学方法进行统计、分析。纳入近年中国国内报道的西罗莫司临床应用及安全性方面的研究论文,要求其中不良反应报告的病例在性别、年龄、不良反应表现等方面详细具体。排除综述类文献及不良反应报告过于简单的病例资料。 结果与结论：共选入文献15篇,患者500例,其中发生不良反应病例合计468例次。468例不良反应中高脂血症发生最多(182例,占38.9%),其次为感染和发热 (62例,占13.2%),肝功能异常 (54例,占11.5%)等。提示西罗莫司主要不良反应为高脂血症、感染和肝功能异常等,不良反应发生与血药浓度等因素相关,定期监测血药浓度,合理调整用量,可增加西罗莫司应用的有效性及安全性。 关键词：西罗莫司;不良反应;文献分析;肾移植;免疫抑制剂     

Clinical, biochemical, and neuropathologic findings following transplantation of adrenal medulla to the caudate nucleus for treatment of Parkinson's disease

We transplanted autologous adrenal medullary tissue into the caudate nucleus of 3 patients with advanced Parkinson's disease. The 1st patient, a 59-year-old man with parkinsonian symptoms for 15 years, had mild improvement in his motor functioning after the operation. However, his postoperative course was characterized by prolonged drowsiness and complex visual hallucinations. The patient died suddenly 8 months after the transplant, and an autopsy revealed coronary atherosclerosis. Examination of the graft site showed necrotic adrenal medullary tissue surrounded by inflammatory cells. The 2nd patient, a 50-year-old man with a 21-year history of parkinsonian symptoms, improved the most after the procedure. The 3rd patient, a 43-year-old man with 12 years of parkinsonian symptoms, had mild improvement in his motor functioning. CSF homovanillic acid increased postoperatively in the 3 patients, but then returned to preoperative levels in all except the 2nd patient. The anatomic, neurochemical, and physiologic basis for the modest clinical improvement shown in these patients is not yet understood.     

低温保存同种异体带瓣主动脉移植的免疫反应

背景：前期研究证实,液氮低温保存同种异体主动脉能保持其细胞的生物活性和组织结构的完整性,但有关同种异体带瓣主动脉移植的排斥反应研究较少。 目的：观察同种异体带瓣主动脉移植后移植物的免疫指标变化特点。 方法：将低温保存的SD大鼠带瓣主动脉异位植入Wistar大鼠腹主动脉,分组干预：同种异体移植组不进行干预;免疫干预组移植后给予环孢素A;对照组为假移植,将腹主动脉切断后直接吻合。移植后1,2,3,4,5周收集移植动物血清标本,免疫组织化学染色测量MHCⅡ阳性表达,淋巴细胞涂片免疫组织化学测定CD4+、CD8+百分率;应用酶联法测定血清中白细胞介素2,肿瘤坏死因子α水平。 结果与结论：同种异体主动脉移植后1~4周可见MHC显著阳性,CD4+、CD4+/CD8+比值明显增高,血清中白细胞介素2和肿瘤坏死因子α水平明显增高,并在移植后5周内维持较高水平,与对照组比较差异有显著性意义(P < 0.05)。环孢素A对同种异体主动脉移植物内膜增厚、淋巴细胞浸润、MHCⅡ表达、T淋巴细胞激活、白细胞介素2和肿瘤坏死因子α水平的增高具有明显抑制作用(P < 0.05)。说明低温保存同种异体主动脉移植后有明确的免疫排斥反应发生,环孢素A对同种异体移植后的免疫排斥反应有明显抑制作用。     

Cell adhesion molecules in adrenal medulla grafts: enhancement of chromaffin cell L1/Ng-CAM expression and reorganization of extracellular matrix following transplantation

Intracerebral adrenal medulla grafts have been used in human patients as an experimental treatment for Parkinson's disease, based on studies in animal models of this disorder. However, alterations in chromaffin cell properties after transplantation and the factors controlling graft survival are poorly understood. Since cell adhesion molecules (CAMs) are involved in regeneration and development of neural tissue in vivo and in vitro, the present study was undertaken to determine the expression of CAMs in adrenal medulla isografts. Fragments of rat adrenal medulla were implanted into the right lateral ventricle. The majority of grafts survived quite well, for up to 2 months (the longest studied period). The implanted chromaffin cells did not develop extensive processes. The cells retained tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) immunoreactivity, while phenylethanolamine N-methyltransferase (PNMT) expression was decreased. Surviving transplanted chromaffin cells showed enhancement and spreading of surface L1/Ng-CAM expression as compared to normal chromaffin cells in adrenal medulla. The implanted chromaffin cells demonstrated only partial conversion to neuronal phenotypes. These chromaffin cells did not develop extensive processes, but showed an enhancement of L1/Ng-CAM expression. Surviving chromaffin cells were accompanied by reorganization of their closely associated extracellular matrix (ECM). As compared to normal in situ adrenal medulla, graft ECM demonstrated a substantial increase of L1/Ng-CAM and laminin immunoreactivities and a distinct decrease in J1/tenascin expression. Some adrenal medulla grafts degenerated, particularly when misplaced within the host brain parenchyma. In these cases the grafts showed fragmentation of ECM and gradual disappearance of CAMs. These results suggest that surviving adrenal medulla grafts exhibit increased synthesis of certain CAMs by chromaffin cells, which may be involved in interactions between chromaffin cells and the surrounding ECM. It is speculated that both surviving and degenerating adrenal medulla grafts could provide CAMs and ECM components including laminin to host brain and this way contribute to functional effects of grafts.     

Catecholamine content of intracerebral adrenal medulla grafts

The rotational behavior which is produced by substantia nigra lesions can be decreased by adrenal medulla grafts adjacent to the denervated striatum. Perhaps these grafts secrete dopamine that diffuses into the striatum. In the present study, we measured concentrations of catecholamines in adrenal medulla grafts as compared with the normal adrenal medulla. The grafts were found to have high but extremely variable concentrations of dopamine. In hosts with substantia nigra lesions, concentrations of dopamine in the adrenal medulla grafts were decreased. Substantia nigra lesions, however, tended to increase concentrations of epinephrine in the grafts, while norepinephrine and total catecholamine concentrations were not significantly affected. It is concluded that at least some adrenal medulla grafts contain concentrations of dopamine sufficient to account for their behavioral effects.     

Altered reactivity of the rat adrenal medulla

Previous studies have suggested that experimental alterations in adrenomedullary reactivity, i.e., changes in catecholamine release in response to a standard dose of acetylcholine, may be partially accounted for by changes occurring at the level of the adrenal medulla itself, independent of both the central nervous system and the innervation of the adrenal gland. The present study was designed to investigate the morphology of adrenal chromaffin cells in rats subjected to chronic hypoglycemia induced by long acting insulin, and to assess this morphology in terms of associated changes in catecholamine content and release. Surgically isolated, perfused adrenal gland preparations were utilized to characterize the functional release of catecholamines from the adrenal medulla. Pretreatment with long acting insulin resulted in a selective depletion of epinephrine stores and acetylcholine-mediated epinephrine release, but did not appear to significantly affect either the levels or the release of norepinephrine. The biochemical effects of long acting insulin persisted for several days after termination of the treatment, exhibiting a gradual recovery over a period of approximately 5 days. Electron microscopic examination of the adrenal chromaffin cells revealed a progressive degranulation and vacuolization of numerous chromaffin cells followed by a compensatory biosynthetic response and a gradual recovery toward the morphology of control cells. The functional release of catecholamines from adrenal chromaffin cells was further examined in preparations of perfused adrenal slices. Acetylcholine-mediated catecholamine release was significantly decreased in slices of adrenal glands prepared from insulin treated rats when compared with that of control animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Intracerebral adrenal medulla grafts: a review

This review summarizes basic and clinical research on intracerebral adrenal medulla grafts, emphasizing potential applications to Parkinson's disease. Properties of intraventricular and intraparenchymal grafts are described, and cell survival and functional effects are compared. It is clear that adrenal medulla allografts survive poorly in the parenchyma of the corpus striatum and better in the lateral ventricle. Nerve growth factor (NGF) may improve the survival of adrenal medulla grafts. In the absence of added NGF even adrenal medulla grafts in the ventricle survive irregularly, and the factors required for graft survival in the ventricle are not well understood. In the 6-hydroxydopamine-lesioned rat model most evidence suggests, not surprisingly, that adrenal medulla grafts produce functional effects only when they survive. These effects may be related to production of catecholamines by the transplanted cells. In addition, adrenal medulla grafts may produce trophic effects on host brain. These effects are most evident in animals with MPTP-induced damage to dopaminergic systems and may be nonspecific, possibly related in part to the brain injury that is induced by graft implantation. Trophic effects may contribute to the functional effects of adrenal medulla grafts: For intraparenchymal grafts, trophic effects that do not require cell survival may contribute small functional changes, while additional behavioral effects may require substantial chromaffin cell survival. The evidence for direct dopamine-mediated effects as compared to trophic mechanisms of action for these grafts in animal models for Parkinson's disease is presented. Clinical studies of adrenal medulla grafts in human patients are examined and compared in detail. When inspected closely, the various clinical studies are in general agreement on most points, although there are differences in the degree of improvement found, both across different studies and individual patients. It is concluded that some beneficial clinical effects occur, with small to modest changes in most patients and substantial improvement in a minority of patients. There also seem to be larger or more consistent changes in durations of "on" and "off" times in L-dihydroxyphenylalanine-treated patients. There are substantial side effects, and it is not clear that the clinical changes are sufficient to justify performing adrenal medulla transplantation in human patients as a routine procedure.(ABSTRACT TRUNCATED AT 400 WORDS)     

Organization, fine structure, and viability of the human adrenal medulla: considerations for neural transplantation

Recent reports of adrenal medullary autografts in patients with Parkinson's disease raise several important questions with respect to the cell types actually being transplanted as well as the potential for chromaffin cell banking prior to neural transplantation. In this study, we determined the general morphological characteristics of the human adrenal medulla and assessed factors important for the maintenance of cultured chromaffin cells for later use as transplants. The human adrenal medulla contained islands of cortical cells scattered throughout the gland as well as Schwann cells, nerve endings, endothelial cells, pericytes, isolated ganglionic neurons, and connective tissue elements such as fibroblasts and smooth muscle cells. Because many of these cell types are mitotically active, transplantation of medullary fragments that contain these cells could have far-reaching consequences. One approach that could circumvent the problems arising from multiple cell types in the medulla is differential plating of chromaffin cells prior to transplantation. Differential plating yielded relatively pure populations of chromaffin cells that demonstrated excellent viability if processed within 2 hours after cessation of the gland's circulation. Chromaffin cells cultured in the presence of nerve growth factor exhibited a neuronal phenotype, possessed catecholamine histofluorescence, and displayed tyrosine hydroxylase- and dopamine beta-hydroxylase-like immunoreactivity. The sex and age of the donor did not affect cell viability or morphological characteristics.     

A new focus on interoceptive properties of adrenal medulla

The adrenal medulla is an important part of the sympathoadrenal system. Chromaffin cells of the adrenal medulla respond to a broad spectrum of stressful situations by releasing epinephrine and norepinephrine. Originally, it was accepted that this response is controlled exclusively by central nervous system structures. However, it was also demonstrated that a surgically denervated adrenal medulla can respond directly by secreting epinephrine and norepinephrine during an imbalance of internal environment (hypoglycemia, asphyxia). Published data had documented the innervation of the adrenal medulla by sensory neurons of spinal dorsal root ganglia. In addition, recent data showed that ganglion cells of the adrenal medulla project ascending axons. These data suggested potential transmission of information from the adrenal medulla to the central nervous system regarding metabolic changes in the blood. This paper presents an overview of possible involvement of adrenal medullary chromaffin cells in the detection of changes in the internal environment and in the transmission of this information to the central nervous system.     

Brainstem activation of the adrenal medulla in the cat

Stimulation of the locus coeruleus in 40 cats leads to generalized activation of the sympathetic nervous system characterized by an immediate pressor response which was followed in the post-stimulus period by an increase of 49 ± 10and44 ± 6% in common carotid arterial resistance ipsilateral and contralateral to stimulation, respectively. This later response was not affected by vagotomy or bilateral cervical sympathectomy but was blocked by high spinal cord section. The post-stimulus carotid vasoconstriction response could be entirely eliminated by acute bilateral physiological adrenalectomy in the form of adrenal hilar clamping, an effect which was reversible if the clamps were removed. The carotid vasoconstrictor response was associated with a rise in the circulating level of noradrenaline (260%) and adrenaline (196%), which was prevented by clamping the adrenal hilum. This response was not mediated via the hypothalamus because it persisted in the decerebrate animal, nor was it merely excitation of fibers of passage since it was reproduced by microinjection of glutamate into the locus coeruleus. The response was blocked by phentolamine suggesting it is mediated by α-adrenoceptors. These data represent the first conclusive demonstration that cell bodies in the brainstem are capable of activating the adrenal medulla. This fact is central to our present concept of the organization of the sympatho-adrenal axis.     

Selectivity in the reinnervation of iris and adrenal medulla by superior cervical ganglion after transplantation under the kidney capsule

A previous gross evoked potential study revealed that although multisensory convergence characterizes the entire claustrum, it is a complex, regionally differentiated formation that receives sensory afferents via primary and non-specific pathways. In the present study the responses of individual claustral neurons to somatic stimulation, clicks and flashes were investigated in immobilized cats with and without chloralose anesthesia.Of 397 neurons, 31% were unresponsive to any of the sensory stimuli that were used. The 275 responsive neurons formed a heterogeneous population and were distributed throughout the structure with no apparent order. Ninety percent were activated by somatic stimulation, 48% by click and 5% by flash. Among somatic cells 18% had ‘restricted’ or ‘highly restricted’ fields, 38% had widespread fields and 44% responded to more than one sensory modality. All somatic cell types were driven by mechanical stimulation of the skin or of deep tissues, and no correlation as found between the cell type and a given stimulus type. Latency distribution studies of somatic cells confirmed the macrophysiological subdivision of the structure into anterior, intermediate and posterior regions.Among auditory neurons, 17% responded only to a click and 83% were multisensory. In contrast to the somatic response, click-evoked response properties were determined by the cell type irrespective of the recording site. Though flash was not tested for in all convergent units, only two units were encountered which were activated by flash alone. In total, 75% of the responsive cells showed heterotopic or heterosensory convergence properties, while 25% were specific. Interaction between pairs of different stimuli varied from neuron to neuron and complex interaction patterns were observed. These data confirm the functional non-homogeneity of the claustrum and indicate that sensory afferents from various origins are distributed differently inside the structure and terminate in different combinations upon individual neurons.Comparison of animals with and without chloralose showed that the neuronal populations were similar in regard to overall responsiveness and the respective proportions of convergent and specific neurons. Response types, however, were markedly different in the two preparations. Under chloralose excitatory responses were rather stereotypical and inhibitory effects were difficult to assess due to the silence of the majority of cells. Without chloralose, excitatory responses were long and complex and inhibitory responses (either initially or following excitation) were predominant. In both preparations, spontaneous firing rates were low and rhythmic responses, as well as the driving up of unresponsive neurons by repetitive stimulation, were observed.The data presented are discussed with regard to the functional significance of the claustrum in sensory perception and in regulating non-sensory functions.     

Expression of Fos protein in adrenal preganglionic neurons following chemical stimulation of the rostral ventrolateral medulla of the rat

The ventrolateral medulla is known to be involved in the regulation of arterial blood pressure, especially via its connections with sympathetic preganglionic neurons (SPNs) mainly located in the intermediolateral nucleus of the spinal cord. It has been shown that stimulation of the rostral part of the ventrolateral medulla (RVLM) elicits a release of catecholamines from the adrenal medulla. The aim of the present study was to demonstrate the existence of a functional pathway between the RVLM and adrenal SPNs using the combination of a retrograde tract tracing technique (cholera toxin B subunit) with the immunohistochemical detection of Fos protein following the chemical stimulation of RVLM. The data obtained showed that: (1) chemical stimulation of the RVLM induced Fos immunoreactivity in the intermediolateral nucleus and particularly in SPNs projecting to the adrenal medulla; (2) along the thoracic segments T2-T12, 26.1% of retrogradely identified adrenal SPNs were Fos-immunoreactive with the greatest percentage (30.9%) in the T8 segment. These results favored a functional control of the RVLM on adrenal SPNs which may contribute to a substantial activation of the cardiovascular system via the release of adrenal catecholamines.     

Chromaffin cell proliferation in the adult rat adrenal medulla

Epinephrine and norepinephrine-containing chromaffin cells proliferate in the adrenal glands of normal adult rats throughout life. Moreover, their rate of proliferation is markedly increased by short-term administration of reserpine, one of many agents which in long-term experiments are associated with the development of adrenal medullary tumors. Current data suggest that chromaffin cell proliferation in the adult rat adrenal is mediated by the interaction of neurogenic and hormonal signals. Reserpine is known to directly deplete catecholamine stores, and to reflexively increase the activity of the splanchnic nerve endings innervating the adrenal medulla to stimulate both secretion and synthesis of catecholamines and other secretory granule constituents. Its effect on chromaffin cell proliferation suggests that the same signals may regulate chromaffin cell number to meet physiological needs. The reserpine model might shed light on signal transduction mechanisms which normally promote or prevent proliferation of chromaffin cells and of other neuroendocrine cells during development or in adult life, and on ways in which such mechanisms are altered in the course of the development and progression of tumors. It also suggests the possibility that chromaffin cells might be propagated in vitro for use in basic biological studies or in transplants for the treatment of Parkinson's disease.     

GABAA and GABAB sites in bovine adrenal medulla membranes

The effect of several ligands and Ca2+ ions on [3H]GABA binding to bovine adrenal medulla membranes was investigated. Without any blockade, the [3H]GABA binding showed two components, one of low affinity (Kd = 139 +/- 22 nM and Bmax = 3.2 +/- 0.4 pmol/mg protein) and the other of high affinity (Kd = 41 +/- 6 nM and Bmax = 0.35 +/- 0.26 pmol/mg protein). Muscimol specifically blocked low-affinity sites, and (-)baclofen blocked high-affinity components. Ca2+ ions were strictly necessary for maximum binding to high-affinity sites, whereas they did not significantly affect sites of the lower affinity. These results show that the bovine adrenal medulla has a GABAA receptor population of low affinity together with a GABAB receptor of high affinity.     

Involvement of adrenal medulla grafts in the open field behavior

Immunohistochemical and behavioral techniques were used to study the effects of adrenal medulla grafts, implanted in striatum after bilateral kainic acid (KA) lesions of this structure, on the open field behavior of mice. KA-induced behavioral changes in leaning, grooming and locomotor activity of the open field test were significantly improved after grafting of the adrenal medulla, and in some respects, fully restored. Immunohistochemical identification showed that grafts contained neuron-like cells with a tyrosine hydroxylase (TH), phenylethanolamine N-methyltransferase, gamma-aminobutyric acid (GABA), choline acetyltransferase (ChAT), and enkephalin-like immunostainings. A likely interpretation of this complex pattern of results is that adrenal medullary grafts may restore the deficits of GABAergic neurons which in turn reverse the abnormalities in emotionality and locomotion. Neurobiologically, these behavioral improvements probably involve GABAergic and catecholaminergic factors of adrenal medulla grafts, although other neuroactive substances, such as acetylcholine and enkephalins, cannot be excluded.     

Histamine-immunoreactive endocrine cells in the adrenal medulla of the rat

Histamine is widely distributed in various mammalian tissues^5,13,14 and it has been shown that histamine is located in mast cells as well as in other structures^2,5,25,31,36. Biochemical evidence has been presented that histamine acts as a neurotransmitter in the central nervous system^16,31. Immunohistochemical studies have demonstrated the location of histamine-immunoreactive neuronal cells in both the central^25,36 and peripheral^18,26 nervous system. Biochemical studies have shown that histamine is present in the adrenal gland¹⁰, while the location of histamine in the adrenal medulla is not known. There is pharmacological and biochemical evidence that exogenous histamine affects the catecholamine secretion of the adrenal medulla^32,39.The present study was undertaken to examine the location of histamine in the rat adrenal medulla by an indirect immunofluorescence method⁶ using a specific histamine antiserum. We now report the presence of histamine-immunoreactive endocrine cells in the adrenal medulla of the rat and suggest that histamine is located in the noradrenaline-secreting cells.