细胞钙浓度的变化对周期信号的响应及胞间同步

 钙离子是细胞中一种很重要的第二信使,通常它以钙离子浓度振荡的方式转导多种生理学信息,影响细胞分化、成熟和凋亡等各种生理过程,最终导致生物效应。通过实验研究了细胞钙浓度的变化对周期信号的响应和建立在多细胞模型的基础上,从肝细胞内钙离子振荡的动力学模型出发,以胞间耦合因子作为影响因子,数值分析单细胞、耦合的多细胞下的胞内钙振荡形式。实验结果表明：细胞钙振荡对不同频率和强度的周期信号的响应是不同的：对有的参数周期信号能产生强烈响应,有的不能。数值分析结果表明：细胞间的差异性导致钙振荡不同,胞间耦合影响多细胞钙振荡的同步性。     

耦合细胞体系中多重随机共振对噪声的选择效应

以Hǒfer提出的细胞内钙离子振荡模型为研究对象,考察了噪声诱导的钙振荡信号在一维双向耦合细胞体系中的传递行为,发现当单个细胞受到外噪声扰动时,其输出信号的信噪比（signal—to-noise ratio：SNR）会在两个不同的噪声强度处呈现极大值,表明噪声在细胞中可以诱导出双重随机共振现象．特别重要的是,当多个细胞耦合时,通过耦合作用,第一个细胞中的共振行为可以沿着耦合细胞链向下传递,并表现出不同的特征：离第一个细胞较近和较远的细胞表现出双重随机共振现象,而处于中间位置的细胞则表现出多重随机共振现象．并且,产生多重共振的细胞数目会随着耦合强度的增加而增多．这些现象表明细胞体系可以通过调节耦合强度,借助于多重共振机制,对噪声诱导的细胞信息做出有效的选择．     

基于钙响应钟型曲线构建的钙振荡模型

细胞质钙离子(Ca~(2+))常以浓度振荡变化的方式控制许多生理活动,内质网上的三磷酸肌醇受体(inositol 1,4,5-trisphosphate receptor,IP_3R)通道是产生钙振荡的关键因素,因此研究其振荡机制具有重要意义.虽然钙振荡已被广泛建模,但大部分模型细胞质与内质网中钙振荡的范围与实验观测数据差异较大.通过整合最近的实验结果,基于IP_3R通道活性对细胞质Ca~(2+)浓度依赖的钟型曲线构建了一个新的钙振荡模型.模型结果除了可以很好地模拟实验中关于钙振荡范围的数据,还可重复多种实验现象.对模型进行参数敏感性分析后,进一步采用单参数分岔分析分别研究了两个高敏感参数,即细胞质内Ca~(2+)缓冲蛋白的总浓度和IP_3R通道激活Ca~(2+)的解离常数(K_1)对钙振荡的影响,发现它们对钙信号有相反的作用.对三磷酸肌醇(IP_3)浓度和K_1进行双参数分岔分析的结果表明K_1对IP_3能产生钙振荡的区域及其振幅有重要影响,揭示可以通过改变K_1影响细胞对外界刺激的响应及其钙振荡模式.该研究有助于理解钙振荡机制,并可为后续理论研究提供框架.     

改进双钙库模型的分岔分析

钙离子(Ca~(2+))是细胞内最常用的信号分子,它通过钙振荡的形式调节多种不同的细胞活动。双钙库模型是一个经典的钙振荡模型,它描述了细胞内钙振荡发生的详细机制。但是,该模型中的希尔系数高,不利于数学分析。本文在降低希尔系数的基础上提出了改进模型,并应用定性理论和分岔理论对其进行了数学分析。改进双钙库模型的动力学性质与原始双钙库模型相似,但其钙振荡区间更大,振幅变化更明显,更好地体现了Ca~(2+)编码细胞信号的方式。双参数分岔分析结合参数敏感性分析的结果表明:Ca~(2+)流出细胞的速率常数、钙库上钙释放通道和钙泵的激活常数均可对钙振荡产生明显影响,这些参数的改变可能会引发Ca~(2+)相关的疾病。本研究有助于人们更好地从数学层面研究双钙库模型,对理解钙信号及其相关疾病也有一定帮助。     

模块化自重构机器人最优构形拓扑转换

定义了模块化自重构机器人构形拓扑转换的基本概念,分析总结了最优构形拓扑转换的规律,并采用遗传算法对构形拓扑转换进行优化.结果显示,构形拓扑转换过程中的连接变化操作次数被有效地降低,提高了模块化自重构机器人自重构的效率.     

大鼠胰腺β细胞内葡萄糖钙信号调控机制

用fura-2显微荧光法测量细胞内自由钙浓度[Ca2+]i,在单个大鼠胰腺β细胞上探讨了葡萄糖诱发的[Ca2+]i初期下降相(GIDP)、钙振荡和钙库释放的影响因素.实验中发现,非刺激浓度的葡萄糖(5 mmol/L)仍可导致β细胞特有的GIDP.GIDP与去极化无关,也不受外钙的影响,但能被内质网钙泵抑制剂thapsigargin抑制,提示此过程可能缘于钙库对胞浆Ca2+的摄取.在胞内钙振荡过程中移去胞外液中的Ca2+,振荡会逐渐停止;若施加thapsigargin,钙振荡仍能继续维持,说明钙振荡主要缘于外钙内流,钙库在钙振荡维持中的作用不大.无外钙条件下,葡萄糖诱导的钙库释放能被钠通道阻断剂TTX所抑制,而高钾对钙库的释放作用却不受TTX的影响,这提示膜去极化可以直接诱导钙库释放.因此,细胞膜去极化、外钙内流、胞内钙库的摄取和释放共同协调着β细胞内的Ca2+平衡.     

改进的De Young-Keizer钙振荡模型

钙离子(Ca~(2+))是细胞内重要的信使分子,其通过钙振荡的形式控制很多细胞活动.为了研究钙振荡产生的机制,研究者们构建了较多钙振荡模型,由De Young和Keizer构建的模型(DYK模型)是其中的典型代表.但DYK模型有一些不足之处,其最主要的问题是Ca~(2+)振荡的振幅与最新实验数据相差较大.该文依据对DYK模型参数敏感性分析的结果,调整了其中的3个参数,并考虑Ca~(2+)缓冲蛋白的作用,提出改进模型.改进模型的时间序列和以[IP_3]为分岔参数分析的结果显示:细胞质和内质网Ca~(2+)振荡的范围符合实验测量结果.此外,微调改进模型的3个参数可以使钙信号编码方式呈现调幅、调频和调幅加调频混合模式.该研究不仅可使人们从理论方面理解钙振荡产生的机制及钙信号的编码方式,而且可为后续研究提供能真实反映实验结果的数学模型及理论框架.     

2型糖尿病鼠胰腺β细胞中SERCA泵的变化对钙振荡波形影响的建模研究

在原内质网Ca2+-ATP酶(SERCA)模型的基础上建立了2型糖尿病鼠胰腺β细胞的SERCA模型,并将两模型分别应用于钙振荡模型里来模拟SERCA泵的变化对胞质钙浓度([Ca2+])的影响.分析结果发现:(1)病鼠胰腺β细胞中的[Ca2+]升高,并且钙振荡周期和幅值增大;(2)细胞中ATP浓度([ATP])降低,ATP振荡周期增大,但幅值变化不明显;(3)内质网(ER)中钙振荡波形的周期增大, 幅值减小.结论:在2型糖尿病鼠胰腺β细胞中,其[Ca2+]相对于正常大鼠明显升高的现象与细胞中SERCA泵转运钙的能力下降有关.     

内噪声对细胞体系钙振荡的积极作用

通过构建一个细胞内钙振荡的介观随机模型,采用化学朗之万方程,研究了内噪声对细胞内钙振荡的影响.研究结果发现,当体系处于由宏观确定性方程所确定的Hopf分叉点附近,并且此时体系的确定性动力学行为不发生振荡时,如果考虑内噪声的作用,就会有随机的钙振荡发生,并且这种钙振荡的规则度会随着内噪声强度的变化出现一个极大值,表明了内噪声随机共振的发生.     

除草剂乙草胺抑制大鼠肝脏细胞肾上腺素能受体所介导的胞浆钙振荡（英文）

田间除草剂乙草胺施用之后非常容易进入地表水和地下水,河流溪水中乙草胺浓度可达纳摩尔水平,从而对水生生物产生长期影响.因职业关系而产生的皮肤暴露和吸入,可导致人血液乙草胺浓度达到微摩尔水平.对乙草胺体内动力学的研究,发现肝脏是乙草胺毒理作用的主要靶器官.已知在肝脏细胞多种生理功能中,钙离子发挥重要作用.本文研究在新鲜分离的大鼠肝脏细胞,乙草胺对肾上腺素能受体所介导胞浆钙振荡的可能影响.实验发现低浓度乙草胺(1、10μmol·L^-1)对苯丙肾上腺素所诱导钙振荡没有影响,但是高浓度(50、100、200μmol·L^-1)乙草胺在有些肝脏细胞可逆性抑制苯丙肾上腺素所诱导的胞浆钙振荡.在苯丙肾上腺素2次串联刺激之间短暂加入乙草胺(1、10、100μmol·L^-1),乙草胺对肝脏细胞基础钙浓度没有影响,也不影响第2次苯丙肾上腺素刺激所引发胞浆钙振荡.细胞免疫化学研究发现在新鲜分离的大鼠肝脏不同细胞,α1肾上腺素能受体密度存在明显差异.对固定的大鼠肝脏切片进行组织免疫化学检测,发现α1肾上腺素能受体在肝脏小叶的密度梯度分布：α1肾上腺...     

A model of calcium signaling and degranulation dynamics induced by laser irradiation in mast cells

Recent experiments show that calcium signaling and degranulation dynamics induced by low power laser irradiation in mast cells must rely on extracellular Ca^2＋ influx. An analytical expression of Ca^2＋ flux through TRPV4 cation channel in response to interaction of laser photon energy and extracellular Ca^2＋ is deduced, and a model characterizing dynamics of calcium signaling and degranulation activated by laser irradiation in mast cells is established. The model indicates that the characteristics of calcium signaling and degranulation dynamics are determined by interaction between laser photon energy and Ca^2＋ influx. Extracellular Ca^2＋ concentration is so high that even small photon energy can activate mast cells, thus avoiding the possible injury caused by laser irradiation with shorter wavelengths. The model predicts that there exists a narrow parameter domain of photon energy and extracellular Ca^2＋ concentration of which results in cytosolic Ca^2＋ limit cycle oscillations, and shows that PKC activity is in direct proportion to the frequency of Ca^2＋ oscillations. With the model it is found that sustained and stable maximum plateau of cytosolic Ca^2＋ concentration can get optimal degranulation rate. Furthermore, the idea of introducing the realistic physical energy into model is applicable to modeling other physical signal transduction systems.     

A defined range of guard cell calcium oscillation parameters encodes stomatal movements.

Oscillations in cytosolic calcium concentration ([Ca2+]cyt) are central regulators of signal transduction cascades, although the roles of individual [Ca2+]cyt oscillation parameters in regulating downstream physiological responses remain largely unknown. In plants, guard cells integrate environmental and endogenous signals to regulate the aperture of stomatal pores and [Ca2+]cyt oscillations are a fundamental component of stomatal closure. Here we systematically vary [Ca2+]cyt oscillation parameters in Arabidopsis guard cells using a 'calcium clamp' and show that [Ca2+]cyt controls stomatal closure by two mechanisms. Short-term 'calcium-reactive' closure occurred rapidly when [Ca2+]cyt was elevated, whereas the degree of long-term steady-state closure was 'calcium programmed' by [Ca2+]cyt oscillations within a defined range of frequency, transient number, duration and amplitude. Furthermore, in guard cells of the gca2 mutant, [Ca2+]cyt oscillations induced by abscisic acid and extracellular calcium had increased frequencies and reduced transient duration, and steady-state stomatal closure was abolished. Experimentally imposing [Ca2+]cyt oscillations with parameters that elicited closure in the wild type restored long-term closure in gca2 stomata. These data show that a defined window of guard cell [Ca2+]cyt oscillation parameters programs changes in steady-state stomatal aperture.     

Alpha-haemolysin of uropathogenic E. coli induces Ca2+ oscillations in renal epithelial cells

Pyelonephritis is one of the most common febrile diseases in children. If not treated appropriately, it causes irreversible renal damage and accounts for a large proportion of end stage renal failures. Renal scarring can occur in the absence of inflammatory cells, indicating that bacteria may have a direct signalling effect on renal cells. Intracellular calcium ([Ca2+]i) oscillations can protect cells from the cytotoxic effects of prolonged increases in intracellular calcium. However, no pathophysiologically relevant protein that induces such oscillations has been identified. Here we show that infection by uropathogenic Escherichia coli induces a constant, low-frequency oscillatory [Ca2+]i response in target primary rat renal epithelial cells induced by the secreted RTX (repeats-in-toxin) toxin alpha-haemolysin. The response depends on calcium influx through L-type calcium channels as well as from internal stores gated by inositol triphosphate. Internal calcium oscillations induced by alpha-haemolysin in a renal epithelial cell line stimulated production of cytokines interleukin (IL)-6 and IL-8. Our findings indicate a novel role for alpha-haemolysin in pyelonephritis: as an inducer of an oscillating second messenger response in target cells, which fine-tunes gene expression during the inflammatory response.     

稀土对红豆杉细胞内游离钙含量的影响

采用Ｆｕｒａ－２／ＡＭ双波长荧光分光光度计测定了稀土不同作用时间和不同剂量,以及红豆杉细胞不同生长时期和在钙通道阻断剂存在的情况下,红豆杉细胞内游离钙离子浓度的变化,实验结果表明,稀土对红豆杉细胞内钙离子浓度的影响随作用时间和剂理的不同而改变,细胞不同的生长时期对稀土的敏感程度不同,引外发现稀土引起细胞内钙离子浓度的振荡是由于胞内钙库释放和胞外钙内流所致。     

Oscillations of cytosolic Ca2+ in pituitary cells due to action potentials

Electrical activity in non-neuronal cells can be induced by altering the membrane potential and eliciting action potentials. For example, hormones, nutrients and neurotransmitters act on excitable endocrine cells. In an attempt to correlate such electrical activity with regulation of cell activation, we report here direct measurements of cytosolic free Ca2+ changes coincident with action potentials. This was achieved by the powerful and novel combination of two complex techniques, the patch clamp and microfluorimetry using fura 2 methodology. Changes in intracellular calcium concentration were monitored in single cells of the pituitary line GH3B6. We show that a single action potential leads to a marked transient increase in cytosolic free calcium. The size of these short-lived maxima is sufficient to evoke secretory activity. The striking kinetic features of these transients enabled us to identify oscillations in intracellular calcium concentration in unperturbed cells resulting from spontaneous action potentials, and hence provide an explanation for basal secretory activity. Somatostatin, an inhibitor of pituitary function, abolishes the spontaneous spiking of free cytosolic Ca2+ which may explain its inhibitory effect on basal prolactin secretion. Our data therefore demonstrate that electrical activity can stimulate Ca2+-dependent functions in excitable non-neuronal cells.     

Sphingosine-1-phosphate induces Ca2+ mobilization via TRPC6 channels in SH-SY5Y cells and hippocampal neurons

Sphingosine-1-phosphate (S1P) is a widely expressed biologically active sphingolipid that plays an important role in cell differentiation, migration, proliferation, metabolism and apoptosis. S1P activates various signaling pathways, some of which evoke Ca2+ signals in the cytosol. Few studies have focused on the mechanism by which S1P evokes Ca2+ signals in neurons. Here, we show that S1P evokes global Ca2+ signals in SH-SY5Y cells and hippocampal neurons. Removal of extracellular calcium largely abolished the S1P-induced increase in intracellular Ca2+, suggesting that the influx of extracellular Ca2+ is the major contributor to this process. Moreover, we found that S1P-induced Ca2+ mobilization is independent of G protein-coupled S1P receptors. The TRPC6 inhibitor SAR7334 suppressed S1P-induced calcium signals, indicating that the TRPC6 channel acts as the downstream effector of S1P. Using patch-clamp recording, we showed that S1P activates TRPC6 currents. Two Src tyrosine kinase inhibitors, Src-I1 and PP2, dramatically inhibited the activation of TRPC6 by S1P. Taken together, our data suggest that S1P activates TRPC6 channels in a Src-dependent way to induce Ca2+ mobilization in SH-SY5Y cells and hippocampal neurons.     

钙离子信号及细胞调控信号网络动力学

钙离子(Ca²⁺)是细胞内广泛存在的一种重要的第二信使,参与并控制着几乎所有的生命活动过程.细胞信号分子网络对细胞正常和病理生理活动过程进行着精密调控,确保细胞各项生理功能有序地进行.本文综述了近些年本课题组关于细胞内钙信号及细胞信号网络动力学模型方面的研究进展,包括集团化钙离子通道释放局域钙信号、细胞全局钙波信号、内质网和线粒体钙微域调控钙信号和钙信号调控细胞凋亡信号网络动力学,以及细胞信号调控网络动力学等.这些理论工作为研究钙信号和蛋白质信号网络调控细胞复杂生命过程的动力学机制提供了方向和思路.     

环腺苷二磷酸核糖（cADPR）类似物的合成与诱导钙释放活性的研究

Ca²⁺信号传导通路是生物体内重要的胞内信号传导途径之一。局部钙信号主要来源于细胞内钙库释放,而这些钙信号受到各种第二信使的控制和Ca²⁺通道蛋白的调节。环腺苷二磷酸核糖（cADPR）作为烟酰胺腺嘌呤二核苷酸（NAD⁺）的代谢物,发现于1987年,是一种信号传导分子,它广泛存在于各种生物系统中,通过介导兰诺定( RyR) 受体调节钙动员活性。研究cADPR以及具有不同生物活性的类似物之间的构效关系是探究分子内钙释放机制的主要手段,另外,一些结构新颖的拮抗剂和激动剂可以作为研究细胞系统复杂机制的研究工具。作者概括性地介绍了cADPR结构类似物——N¹-乙氧基甲基-环肌苷-5'-二磷酸核糖（cIDPRE）和N¹-[(磷酰基-O-乙氧基)-甲基-N⁹-[(磷酰基-O-乙氧基)-甲基-次黄嘌呤-环磷酸焦酯（cIDPDE）的合成与性质。这两种类似物cIDPDE和cIDPRE可作为研究完整细胞钙信号系统的膜透性激动剂。     

Oscillations of intracellular Ca2+ in mammalian cardiac muscle

Contraction of cardiac muscle depends on a transient rise of intracellular calcium concentration ([Ca2+]i) which is initiated by the action potential. It has, however, also been suggested that [Ca2+]i can fluctuate in the absence of changes in membrane potential. The evidence for this is indirect and comes from observations of (1) fluctuations of contractile force in intact cells, (2) spontaneous cellular movements, and (3) spontaneous contractions in cells which have been skinned to remove the surface membrane. The fluctuations in force are particularly prominent when the cell is Ca2+-loaded, and have been attributed to a Ca2+-induced Ca2+ release from the sarcoplasmic reticulum. In these conditions of Ca2+-loading the normal cardiac contraction is followed by an aftercontraction which has been attributed to the synchronization of the fluctuations. The rise of [Ca2+]i which is thought to underlie the aftercontraction also produces a transient inward current. This current, which probably results from a Ca2+-activated nonspecific cation conductance, has been implicated in the genesis of various cardiac arrhythmias. However, despite the potential importance of such fluctuations of [Ca2+]i their existence has, so far, only been inferred from tension measurements. Here we present direct measurements of such oscillations of [Ca2+]i.