肾上腺素和去甲肾上腺素对离体大鼠颗粒细胞雌二醇生成的影响

本工作采用离体孵育大鼠颗粒细胞的方法,观察了肾上腺素和去甲上腺素对颗粒细胞雌二醇生成的影响,为进一步探讨其作用机制,实验还观察了β－肾上腺素能受体阻断剂心得安,外源性ｃＡＭＰ和腺苷酸环化酶激动剂对肾上腺素和去甲肾上腺素诱导颗粒细胞雌二醇生成的影响,结果发现,肾上腺素和去甲肾上腺素可明显促进离体大鼠颗粒细胞雌二醇的生成,此作用可被心得安阻断,外源性ｃＡＭＰ和Ｆｏｒｓｋｏｌｉｎ可促进肾上腺素和去甲肾上     

Nogo-A及其受体在中枢神经系统发育过程中作用及机制的研究进展

Nogo-A及其受体在成年哺乳动物的中枢神经系统(CNS)中,尤其是在中枢神经系统损伤及修复过程中的作用及机制已经被广泛而深入的研究,但是它们在CNS发育中的扮演的角色却了解甚少。新近研究表明,Nogo-A在CNS发育过程中神经前体细胞分化及迁移,轴突的生长及可塑性的变化以及少突胶质细胞前体细胞分化和成髓鞘化等过程中发挥着重要的作用。     

Nogo-A及其受体在中枢神经系统发育过程中作用及机制的研究进展

Nogo-A及其受体在成年哺乳动物的中枢神经系统(CNS)中,尤其是在中枢神经系统损伤及修复过程中的作用及机制已经被广泛而深入的研究,但是它们在CNS发育中的扮演的角色却了解甚少.新近研究表明,Nogo-A在CNS发育过程中神经前体细胞分化及迁移,轴突的生长及可塑性的变化以及少突胶质细胞前体细胞分化和成髓鞘化等过程中发挥着重要的作用.     

老年大鼠血管α1肾上腺素受体及其亚型的改变

本工作用离体与整体实验方法,研究了老年(18月龄)与年轻(3月龄)大鼠血管中 α_1肾上腺素受体储备和 α_1(?)与 α_(1b)亚型比值的差别。在离体实验中用机械方法分离血管,用 Krebs 溶液灌流,在灌流液中加入 α_2和 β肾上腺素受体拮抗剂,然后用去甲肾上腺素(NE)激动 α_1受体。结果显示;老年大鼠主动脉、肾动脉和肠系膜动脉由去甲肾上腺素(NE)引起的最大收缩反应与年轻大鼠无显著差别,但浓度-效应曲线显著右移,功能性解离常数 K_A 值不变,而 K_A 与EC_(50)的比值减小。此外,在老年大鼠主动脉和肠系膜动脉血管,选择性 α_(1b)亚型拮抗剂 CEC对 NE 引起的缩血管效应的阻断作用显著减弱,硝苯吡啶(选择性阻断 α_(1a) 亚型的效应)对 NE 缩血管效应的阻断作用显著增强。整体实验显示老年大鼠硝苯吡啶的降血压作用比年轻大鼠增强,在用硝苯吡啶的基础上给予苯肾上腺素升血压作用减弱。上述结果提示:与年轻大鼠相比较,老年大鼠 α_(1-) 肾上腺素受体储备减少,α_(1a) 亚型相对 α_(1b)亚型的比率增高。     

β-肾上腺素受体与乳腺癌相关性的研究进展

精神和情绪对于肿瘤的发生、发展具有重要作用。通常,在急性或慢性应激状态下,由交感神经系统介导的正常生理应激机制导致神经递质肾上腺素和去甲肾上腺素释放增多。对于恶性肿瘤患者,大量研究证实交感神经系统主要通过β-肾上腺素能受体途径介导的信号传导通路对肿瘤的进展和转移产生影响。乳腺癌患者多伴有焦虑预后不良而引起肾上腺素及去甲肾上腺素分泌增加,从而可能促进乳腺癌的进展和转移。本文就β-肾上腺素受体及其在其它恶性肿瘤中相关研究,着重探讨β-肾上腺素受体在乳腺癌中表达情况和使用β-肾上腺素受体阻断剂与乳腺癌预后的关系作以下综述,为进一步探究β-肾上腺素受体在乳腺癌中的作用机制提供相关理论依据。     

β-catenin在生后大鼠脑组织的表达及变化

目的观察-βcatenin在成年大鼠脑组织的表达及其在生后发育过程中的变化,探讨其在中枢神经系统(CNS)发育分化中的作用。方法用免疫组织化学定性定位检测-βcatenin在脑组织中时空表达,Western blotting方法半定量检测发育过程中皮层-βcatenin的变化。结果-βcatenin在成年大鼠脑内主要分布于皮层锥体细胞层、海马、室下层及丘脑等区域,阳性细胞多为有突起的神经元样,在密集表达的室下区及丘脑近脑室等处,-βcatenin有明显的细胞核内定位;新生大鼠脑内-βcatenin呈散在分布,P3至P20-βcatenin表达逐步增多,主要存在于新皮层、丘脑及室下层区域,尤其在扣带后皮质、纹状皮质、梨状前皮质等处。老年大鼠脑内-βcatenin的表达分布基本与成年相似,但阳性细胞数量及强度显著低于成年。Western blotting显示-βcatenin在CNS皮层生后发育过程中存在持续表达,表达高峰主要在P3和P21两个时相。结论-βcatenin在CNS生后的时空表达具有相对固定的模式,同时呈现明显的部位和表达量的变化,提示-βcatenin与CNS分化发育、特别是神经发生存在密切关系。     

热应激时大鼠肺中肾上腺素能受体的变化及其调节机理

研究了急性热应激致大鼠肛温达42℃时即刻、持续15min及持续15min并在室温恢复4h后三种状态下肺肾上腺素能受体(α,β)的动态改变及其相应配基去甲肾上腺素(NE),肾上腺素(E)含量的改变。同时测定了肺中磷脂酶A_2(PLA_2)活性的变化。结果表明,急性热应激可使大鼠肺中肾上腺素能受体发生改变,内源性配基NE、E及PLA_2的活性也相应发生改变。NE和/或E含量以及PLA_2活性的改变在肾上腺素能受体变化的过程中可能起重要的调制作用。     

CXCL12/CXCR4轴对NPC调节的研究进展

基质细胞衍生因子1α(SDF-1α/CXCL12)属于趋化因子CXC家族,与其受体CXCR4组成的CXCL12/CXCR4轴,在大脑生理和病理状态下都发挥着重要作用。CXCL12能与神经祖细胞(NPC)表面上的受体CXCR4结合,从而激活CXCR4下游不同的信号通路,参与调节NPC静息、激活、增殖、迁移和分化等活动。在中枢神经系统(CNS)疾病发生后,大脑中CXCL12会激活内源的NPC,促进NPC增殖并迁移至病灶区域,最终分化为神经元并整合入神经系统,促进神经功能恢复。深入理解CNS疾病时期CXCL12/CXCR4轴对NPC调控作用,对内源性和外源性的NPC应用于CNS疾病具有重要意义。现主要对CXCL12/CXCR4轴调控NPC活动的作用机制及相关信号通路进行综述。     

肾上腺素受体的生物进化

肾上腺素受体在生物进化的过程中最先出现的是β－肾上腺素受体,此后才出现了α－肾上腺素受体。肾上腺素受体亚型的多样化分化是伴随大脑神经系统的发育而进行的,提示肾上腺素受体的各亚型的进一步分化很有胆对高级神经系统的精细调节功能不断趋于完善的一种适应,同时也预示可能还有其它肾上腺素受体亚型存在。     

用a1D受体高表达细胞膜色谱研究9种配体的生物亲和作用

为了增加细胞膜色谱法的选择性和特异性,用受体高表达细胞膜色谱法研究9种a1-肾上腺素受体配体与a1D-肾上腺素受体亚型(a1D-AR)的生物亲和作用.培养稳定表达a1D-AR的HEK293细胞株,制备细胞膜固定相,应用受体高表达细胞膜色谱法研究不同配体与a1D-AR的结合情况.结果表明:9种不同的a1-肾上腺素受体配体与大鼠a1D-AR的亲和顺序为:哌唑嗪,BMY7378,酚妥拉明,羟甲唑啉,5-甲基乌拉地尔,去甲肾上腺素,苯肾上腺素,甲氧明,RS-17053.受体高表达细胞膜色谱法是一种特异、可靠的生物亲和色谱方法,可用于a1D-AR亚型选择性药物的高效筛选.     

Neurotransmitters as early signals for central nervous system development

During brain ontogenesis, the temporal and spatial generation of the different types of neuronal and glial cells from precursors occurs as a sequence of successive progenitor stages whose proliferation, survival and cell-fate choice are controlled by environmental and cellular regulatory molecules. Neurotransmitters belong to the chemical microenvironment of neural cells, even at the earliest stages of brain development. It is now established that specific neurotransmitter receptors are present on progenitor cells of the developing central nervous system and could play, during neural development, a role that has remained unsuspected until recently. The present review focuses on the occurrence of neurotransmitters and their corresponding ligand-gated ion channel receptors in immature cells, including neural stem cells of specific embryonic and neonatal brain regions. We summarize in vitro and in vivo data arguing that neurotransmitters could regulate morphogenetic events such as proliferation, growth, migration, differentiation and survival of neural precursor cells. The understanding of neurotransmitter function during early neural maturation could lead to the development of pharmacological tools aimed at improving adult brain repair strategies.     

Immunoblot identification of glial fibrillary acidic protein in rat sciatic nerve,brain, and spinal cord during development

The appearance of the glial fibrillary acidic protein (GFAP) during embryonic and postnatal development of the rat brain and spinal cord and in rat sciatic nerve during postnatal development was examined by the immunoblot technique. Cytoskeletal proteins were isolated from the central and peripheral nervous system and separated by SDS slab gel electrophoresis or two-dimensional gel electrophoresis. Proteins from the acrylamide gels were transferred to nitrocellulose sheets which were treated with anti-bovine GFAP serum and GFAP was identified by the immunoblot technique. GFAP was present in the embryonic rat brain and spinal cord at 14 and 16 days of gestation respectively. The appearance of GFAP at this stage of neural development suggests that the synthesis of GFAP may be related to the proliferation of radial glial cells from which astrocytes are derived. It is also feasible that GFAP provides structural support for the radial glial cell processes analogous to its role in differentiated astrocytes. GFAP was found to be present in rat sciatic nerves at birth and at all subsequent stages of development. These results indicate that some cellular elements in the rat sciatic nerve, such as Schwann cells, are capable of synthesizing GFAP which is immunochemically indistinguishable from its counterpart in the central nervous system. Thus it appears that GFAP is present both in the central and peripheral nervous system of the rat when the glial cells synthesizing GFAP are still undergoing differentiation.     

The 13th J. A. F. Stevenson memorial lecture. Sexual differentiation of the brain: possible mechanisms and implications

The mammalian brain appears to be inherently feminine and the action of testicular hormones during development is necessary for the differentiation of the masculine brain both in terms of functional potential and actual structure. Experimental evidence for this statement is reviewed in this discussion. Recent discoveries of marked structural sex differences in the central nervous system, such as the sexually dimorphic nucleus of the preoptic area in the rat, offer model systems to investigate potential mechanisms by which gonadal hormones permanently modify neuronal differentiation. Although effects of these steroids on neurogenesis and neuronal migration and specification have not been conclusively eliminated, it is currently believed, but not proven, that the principle mechanism of steroid action is to maintain neuronal survival during a period of neuronal death. The structural models of the sexual differentiation of the central nervous system also provide the opportunity to identify sex differences in neurochemical distribution. Two examples in the rat brain are presented: the distribution of serotonin-immunoreactive fibers in the medial preoptic nucleus and of tyrosine hydroxylase-immunoreactive fibers and cells in the anteroventral periventricular nucleus. It is likely that sexual dimorphisms will be found to be characteristic of many neural and neurochemical systems. The final section of this review raises the possibility that the brain of the adult may, in response to steroid action, be morphologically plastic, and considers briefly the likelihood that the brain of the human species is also influenced during development by the hormonal environment.     

F-spondin promotes nerve precursor differentiation

Cells in the developing nervous system secrete a large number of proteins that regulate the migration and differentiation of their neighbors. It is shown here that a clonal central nervous system cell line secretes a protein that causes both a rat hippocampal progenitor cell line and primary cortical neural cells to differentiate into cells with the morphological and biochemical features of neurons. This protein was identified as F-spondin. Analysis of F-spondin isoforms secreted from transfected cells shows that the core protein without the thrombospondin type 1 repeats is sufficient to promote neuronal differentiation when adsorbed to a surface. F-spondin can also inhibit neurite outgrowth while allowing the expression of nerve-specific proteins when present in a soluble form at high concentrations. Therefore, F-spondin can alter cell differentiation in multiple ways, depending upon its concentration and distribution between substrate-attached and soluble forms.     

IKAP/Elp1 is required in vivo for neurogenesis and neuronal survival, but not for neural crest migration

Familial Dysautonomia (FD; Hereditary Sensory Autonomic Neuropathy; HSAN III) manifests from a failure in development of the peripheral sensory and autonomic nervous systems. The disease results from a point mutation in the IKBKAP gene, which encodes the IKAP protein, whose function is still unresolved in the developing nervous system. Since the neurons most severely depleted in the disease derive from the neural crest, and in light of data identifying a role for IKAP in cell motility and migration, it has been suggested that FD results from a disruption in neural crest migration. To determine the function of IKAP during development of the nervous system, we (1) first determined the spatial-temporal pattern of IKAP expression in the developing peripheral nervous system, from the onset of neural crest migration through the period of programmed cell death in the dorsal root ganglia, and (2) using RNAi, reduced expression of IKBKAP mRNA in the neural crest lineage throughout the process of dorsal root ganglia (DRG) development in chick embryos in ovo. Here we demonstrate that IKAP is not expressed by neural crest cells and instead is expressed as neurons differentiate both in the CNS and PNS, thus the devastation of the PNS in FD could not be due to disruptions in neural crest motility or migration. In addition, we show that alterations in the levels of IKAP, through both gain and loss of function studies, perturbs neuronal polarity, neuronal differentiation and survival. Thus IKAP plays pleiotropic roles in both the peripheral and central nervous systems.     

A unified hypothesis on the lineage of neural stem cells

For many years, it was assumed that neurons and glia in the central nervous system were produced from two distinct precursor pools that diverged early during embryonic development. This theory was partially based on the idea that neurogenesis and gliogenesis occurred during different periods of development, and that neurogenesis ceased perinatally. However, there is now abundant evidence that neural stem cells persist in the adult brain and support ongoing neurogenesis in restricted regions of the central nervous system. Surprisingly, these stem cells have the characteristics of fully differentiated glia. Neuroepithelial stem cells in the embryonic neural tube do not show glial characteristics, raising questions about the putative lineage from embryonic to adult stem cells. In the developing brain, radial glia have long been known to produce cortical astrocytes, but recent data indicate that radial glia might also divide asymmetrically to produce cortical neurons. Here we review these new developments and propose that the stem cells in the central nervous system are contained within the neuroepithelial --> radial glia --> astrocyte lineage.     

The distribution of heat shock proteins in the nervous system of the unstressed mouse embryo suggests a role in neuronal and non-neuronal differentiation

Heat shock proteins (Hsps) act as molecular chaperones and are generally constitutively expressed in the absence of stress. Hsps are also inducible by a variety of stressors whose effects could be disastrous on the brain. It has been shown previously that Hsps are differentially expressed in glial and neuronal cells, as well as in the different structures of the brain. This differential expression has been related to specific functions distinct from their general chaperone function, such as intracellular transport. We investigated here the constitutive expression of 5 Hsps (the small Hsp, Hsp25, the constitutive Hsc70 and Hsp90beta, the mainly inducible Hsp70 and Hsp90alpha), and of a molecular chaperone, TCP-1alpha during mouse nervous system development. We analyzed, by immunohistochemistry, their distribution in the central nervous system and in the ganglia of the peripheral nervous system from day 9.5 (E9.5) to day 17.5 (E17.5) of gestation. Hsps are expressed in different cell classes (neuronal, glial, and vascular). The different proteins display different but often overlapping patterns of expression in different regions of the developing nervous system, suggesting unique roles at different stages of neural maturation. Their putative function in cell remodeling during migration or differentiation and in protein transport is discussed. Moreover we consider Hsp90 function in cell signaling and the role of Hsp25 in apoptosis protection.     

Dynamics of expression of apoptosis-regulatory proteins Bid, Bcl-2, Bcl-X, Bax and Bak during development of murine nervous system

We have used immunohistochemistry and immunoblotting to examine the expression of Bid and four other Bcl-2 family proteins (Bcl-2, Bcl-X, Bax and Bak) in the developing and adult murine central nervous system (CNS). Bid protein is widespread in embryonic and postnatal brain, and its expression is maintained at a high level late into the adulthood. Bid is expressed both in the germ disc, early neural tube, proliferating stem cells of ventricular zones, and in postmitotic, differentiated neurons of the developing central and peripheral nervous system. As the differentiation proceeds, the neurons express higher levels of Bid than the stem cells of the paraventricular zone. Both in embryonic and postnatal life, Bid protein is present in the most vital regions of brain, such as the limbic system, basal ganglia, mesencephalic tectum, Purkinje cells in cerebellum, and the ventral columns of spinal cord. The p15 cleaved form of Bid was detectable in the brain specimens at fetal stages of development, consistent with caspase-mediated activation of this pro-apoptotic Bcl-2 family protein. Among the Bcl-2 family proteins only Bid and Bcl-XL continue to be expressed at high levels in the adult brain.