Urocortin expression in the human central nervous system

OBJECTIVE AND STUDY DESIGN: Urocortin is a recently identified neuropeptide of the corticotrophin-releasing factor (CRF) family in the mammalian brain and has been demonstrated to stimulate ACTH secretion from pituitary cells, but its expression in human brain tissue including the hypothalamus has not been examined. In this study, we first examined urocortin expression in the hypothalamus (20 cases) and pituitary stalks (17 cases) of human brain obtained from autopsy using immunohistochemistry and mRNA in situ hybridization. RESULTS: Neither urocortin immunoreactivity nor mRNA hybridization signals were detected in the hypothalami and pituitary stalks while CRF immunoreactivity was detected in the paraventricular nuclei of the hypothalami in 10/20 cases and in nerve fibres of the stalks in 17/17 cases. These results indicate that urocortin does not act on the hypothalamo-pituitary-adrenal axis, at least not in the same manner as CRF in humans. We then examined urocortin expression in various portions of the brain in 7 cases. Both urocortin immunoreactivity and mRNA hybridization were detected in Purkinje cells of the cerebellum and anterior horn cells of the spinal cord in specimens examined. Urocortin expression was, however, variably seen in superior olivary nuclei (two out of six cases examined) and in the Edingar-Westphal nuclei (one out of three cases examined). CONCLUSIONS: The distribution of urocortin in the human central nervous system suggests that urocortin may work as a neurotransmitter like other neuropeptides in the human.     

Monoclonal antibodies distinguish antigenically discrete neuronal types in the vertebrate central nervous system.

Eight hundred hybridoma lines were generated from mice immunized with the fixed gray matter of cat spinal cord. Of these lines, 47 were positive when screened immunohistochemically against sections of the cat spinal cord. Twenty-nine lines secreted antibodies that bound to neuronal antigens. Of these, 16 bound to axons only, 8 bound to axons and cell bodies, and 5 bound to cell bodies only. Eighteen lines secreted antibodies that bound to glial cells. Five lines that secreted antibodies that intensely stained spinal cord sections were cloned and screened against other parts of the central nervous system. Each of these five antibodies bound to specific subsets of neurons. For example, in the spinal cord, one antibody (Cat-301) recognized a surface determinant on the dendrites and cell bodies of neurons that, in morphology and location, resemble long-distance projection neurons. A second antibody (Cat-201) recognized an antigen in axons and in the cytoplasm of neuronal cell bodies that may be a subset of those recognized by Cat-301. A third antibody (Cat-101) recognized only axons. The subcellular localization of the antigen recognized by each antibody is the same in all areas of the central nervous system we have examined. The fact that each of the antibodies described here has a restricted distribution in the central nervous system shows that there is a high degree of molecular diversity among vertebrate neurons and that hybridoma technology can be used to explore this diversity. This class of reagents should be a useful addition to the many established techniques for studying the organization of the vertebrate central nervous system.     

Neuroantibodies: molecular cloning of a monoclonal antibody against substance P for expression in the central nervous system.

We present a strategy to study functional and/or developmental processes occurring in the nervous system, as well as in other systems, of mice. This strategy is based on the local expression of specific monoclonal antibodies (mAbs) by cells of the nervous system. As an application of this strategy, we report the cloning of the anti-substance P rat mAb NC1/34HL. Functional substance P-binding antibodies were reconstituted from the cloned variable domains by using vectors for expression in myeloma cells. With these and other vectors a general system for the cloning and expression of mAbs under a series of promoters (of the rat VGF8a gene, the neurofilament light-chain gene, and the methallothionein gene) has been created. The activity of these plasmids was confirmed by expressing the recombinant NC1/34HL mAb in GH3 pituitary cells, PC12 pheochromocytoma cells, and COS cells. DNA from the described constructs can be used to target the expression of the NC1/34HL mAb to the central nervous system of transgenic mice. This procedure will allow us to perturb substance P activity in a controlled way in order to dissect its multiple roles.     

Pathway analysis of primary central nervous system lymphoma

Primary central nervous system (CNS) lymphoma (PCNSL) is a diffuse large B-cell lymphoma (DLBCL) confined to the CNS. A genome-wide gene expression comparison between PCNSL and non-CNS DLBCL was performed, the latter consisting of both nodal and extranodal DLBCL (nDLBCL and enDLBCL), to identify a "CNS signature." Pathway analysis with the program SigPathway revealed that PCNSL is characterized notably by significant differential expression of multiple extracellular matrix (ECM) and adhesion-related pathways. The most significantly up-regulated gene is the ECM-related osteopontin (SPP1). Expression at the protein level of ECM-related SPP1 and CHI3L1 in PCNSL cells was demonstrated by immunohistochemistry. The alterations in gene expression can be interpreted within several biologic contexts with implications for PCNSL, including CNS tropism (ECM and adhesion-related pathways, SPP1, DDR1), B-cell migration (CXCL13, SPP1), activated B-cell subtype (MUM1), lymphoproliferation (SPP1, TCL1A, CHI3L1), aggressive clinical behavior (SPP1, CHI3L1, MUM1), and aggressive metastatic cancer phenotype (SPP1, CHI3L1). The gene expression signature discovered in our study may represent a true "CNS signature" because we contrasted PCNSL with wide-spectrum non-CNS DLBCL on a genomic scale and performed an in-depth bioinformatic analysis.     

Detection of substance P in the central nervous system by a monoclonal antibody.

Peptides with transmitter-like characteristics are being found in many brain areas. The application of immunocytochemical and radioimmunoassay methods has contributed much to the clarification of these neuronal systems. Here we report the development of a rat monoclonal antibody produced by a hybrid myeloma and its application to the study of one of these peptides, substance P. The hybrid clone, isolated after fusion of mouse myeloma cells with hyperimmune rat spleen cells, allowed us to obtain a standardized and permanent source of monoclonal substance P antibodies in a culture cell system. This antibody recognizes the COOH-terminal part of substance P in radioimmunoassay down to 10-20 fmol. It does not crossreact with other known mammalian brain peptides tested. By immunofluorescence the antibody was shown to bind specifically and with a remarkably low background to nerve terminals and cell bodies located in clearly defined nuclear organizations of the central nervous system.     

Cortistatin--functions in the central nervous system

Cortistatin (CST) is a neuropeptide from the somatostatin (SRIF)/urotensin (UII) family named after its predominantly cortical expression and ability to depress cortical activity, which was discovered a decade ago. In vitro assays show CST is able to bind all five cloned somatostatin receptors and shares many pharmacological and functional properties with SRIF. However, distinct from SRIF, CST has been shown to induce slow-wave sleep, reduce locomotor activity, and activate cation selective currents not responsive to somatostatin. Different lines of evidence also indicate that CST, like SRIF, is involved in learning and memory processes. CST-14 may also function as an endogenous anti-convulsant. In addition to its role in cortical synchronization, CST-14 has emerged as an important mediator of immunity and inflammation. This review will cover some of the basic properties of CST in the brain, and will discuss new data on the role of CST in cortical activity.     

Regeneration in the central nervous system

Unlike neonatal axons, mammalian adult axons of the CNS do not regenerate after injury. This developmental loss of regenerative capacity, is correlated with the onset of myelination. Likewise, myelin, or myelin-associated components such as Nogo-A and myelin-associated glycoprotein (MAG) inhibit regeneration from older but not younger neurons. Identification of the molecular events responsible for this developmental loss of regenerative capacity is central to devise strategies to encourage regeneration in adults after injury. Endogenous levels of the cyclic nucleotides cAMP and cGMP have been suggested to determine the neuronal responsiveness to various axonal guidance factors. Elevating cAMP concentrations block Nogo-A or MAG induced inhibition of neurite outgrowth in older neurons, whereas suppressing cAMP levels in young neurons renders them susceptible to Nogo-A and MAG. Interestingly, elevated cAMP levels abrogated the Nogo-A and MAG mediated activation of RhoA and down regulation of Rac1 in adult neurons. In contrast, elevation of cAMP leads to the inactivation of RhoA and prevents activation of downstream effector proteins, while Rac is activated. We therefore conclude that the endogenous neuronal cAMP levels determine the neuronal responsiveness to myelin-associated neurite growth inhibitors by regulating rho GTPase activities.     

31例成人中枢神经系统白血病临床分析

目的：探讨成人非淋巴细胞白血病(ANLL)患者并发中枢神经系统白血病(CNSL)的临床特征。方法：回顾性分析3l例成人CNSL患者的临床资料。结果：3l例成人CNSL患者中，ANLL20例，分别为M2一M5患者。其中3例确诊于发病初期，l7例确诊于巩固期；40岁以下者27例，占87．0％，平均年龄27．48岁，目前死亡15例，失访2例，生存14例。结论：成人ANLL患者并发CNSL发生率高，病情危笃，预后差；临床应重视其预防治疗。     

Intraventricular treatment of relapsed central nervous system lymphoma with the anti-CD20 antibody rituximab

Most patients with primary central nervous system (CNS) lymphoma or systemic non-Hodgkin's lymphoma (NHL) involving the CNS relapse after an initial response to treatment, often presenting with leptomeningeal disease. Since the majority of these lymphomas are B-cell neoplasms expressing the CD20 surface antigen treatment with the chimeric monoclonal antibody (Mab) rituximab might be a new therapeutic option. Here, we report on 6 patients with relapsed CNS B-cell lymphoma who were treated with intraventricular or intrathecal applications of rituximab. One of these cases has already been reported.     

Vasculitis of the central nervous system

Central nervous system (CNS) vasculitis occurs in a variety of clinical settings. Some exhibit a distinct age preference; others a tissue tropism. Most frequently encountered are giant cell arteritis (temporal arteritis) and vasculitis secondary to infections. The CNS may be involved in the systemic vasculitides, and neurologic abnormalities occasionally appear as a presenting manifestation of disease. Isolated angiitis of the CNS, a rare form of vasculitis restricted to the CNS, must be distinguished from other causes of CNS inflammation and from noninflammatory vascular disease. We are learning a great deal about the cellular mechanisms of vascular inflammation in the brain. Some manifestations of the clinical disease result from histologic features of the infiltrate and the size of affected vessel. However, the local consequences of inflammation such as increased coagulation and altered vasomotor tone, as well as the systemic consequences such as activation of the central noradrenergic systems, trigeminovascular system, and hypothalamic pituitary adrenal axis contribute to both pathogenesis of disease and recovery. Two central issues that confront us now are improving the accuracy of the diagnosis (including identifying any underlying infectious causes) and limiting the long-term damage both from disease and its therapies.     

Actinomycosis of the central nervous system

Actinomyces species are rare but treatable causes of CNS infection. Differentiation of actinomycosis from nocardiosis is crucial to the selection of appropriate antimicrobial therapy. A review of 70 cases of CNS actinomycosis was conducted in an effort to characterize clinicopathologic features and identify patients with a high risk of death from infection. Types of lesions included brain abscess (67%), meningitis or meningoencephalitis (13%), actinomycoma (7%), subdural empyema (6%), and epidural abscess (6%). Most infections developed from distant sites (lung, 19 cases; abdomen, four; pelvis, three) or contiguous foci (ear, sinus, and cervicofacial region, 21 cases). For nonmeningitic infection, signs and symptoms were generally those of a space-occupying lesion and were indistinguishable from the manifestations of other pyogenic infections except for a longer interval before diagnosis. Risk factors included dental caries; dental infection; recent tooth extraction; head trauma; gastrointestinal tract surgery; chronic otitis, mastoiditis, or sinusitis; chronic osteomyelitis; tetralogy of Fallot; and actinomyces infection of an intrauterine device. Optimal management combined adequate surgical drainage with prolonged antibiotic therapy (mean duration, 5 months). Overall mortality from treated infection was 28%; 54% of survivors had neurologic sequelae. Features correlated with a poor prognosis were disease onset greater than 2 months before diagnosis and treatment, no antibiotic treatment, no surgery, and needle aspiration drainage of abscess lesions.     

Angiitis of the central nervous system

Angiitis signifies an inflammation of the blood vessels that can cause damage to vessel walls, vascular occlusion, and ischemic injury to organs served by these vessels, and it can occur in the central nervous system. All central nervous system vasculitides but one have been linked to the presence of various underlying disorders. This review covers primary and spinal cord granulomatous angiitis of the central nervous system, as well as secondary angiitis of the central nervous system and its associated underlying disorders.     

Monoclonal antibodies against the Drosophila nervous system.

A panel of 148 monoclonal antibodies directed against Drosophila neural antigens has been prepared by using mice immunized with homogenates of Drosophila tissue. Antibodies were screened immunohistochemically on cryostat sections of fly heads. A large diversity of staining patterns was observed. Some antigens were broadly distributed among tissues; others were highly specific to nerve fibers, neuropil, muscle, the tracheal system, cell nuclei, photoreceptors, or other structures. The antigens for many of the antibodies have been identified on immunoblots. Monoclonal antibodies that identify specific molecules within the nervous system should prove useful in the study of the molecular genetics of neural development.     

Hormone-driven mechanisms in the central nervous system facilitate the analysis of mammalian behaviours

Hormonal effects on behaviours in animals and humans are now well enough understood for general statements about causal steps to be proposed. Facilitation or repression of a given behaviour by a given hormone can depend on the person's genetic and developmental history, on the temporal and spatial parameters of the hormone's administration, on the hormone's metabolism and on the specific receptor isoform available in a given neuron. The gene for oestrogen receptor-alpha is required for an entire chain of behaviours essential for reproduction, from courtship through maternal behaviours. In order to show that it is possible to use endocrine tools to explain a mammalian behaviour, we analysed lordosis behaviour neuronal circuitry as well as the molecular mechanisms of its facilitation by oestrogens. The functional genomics of oestrogenic effects on lordosis arrange themselves in modules for neuronal Growth, Amplification (by progestins), Preparatory behaviours, Permissive actions by hypothalamic neurons, and Synchronization of mating behaviour with ovulation (GAPPS). A related four-gene micronet involving the amygdala and the paraventricular nucleus of the hypothalamus supports social recognition. Underlying all sociosexual behaviour is the fundamental arousal of brain and behaviour. Elementary arousal depends on a bilateral, bidirectional system universal among mammalian brains, and it can be altered by null deletion of the gene for oestrogen receptor-alpha. Future molecular and biophysical studies will specify how hormone effects in the brain change central nervous system state in such a manner as to alter the frequencies of entire sets of behavioural responses.