Sequence and functional expression of the GABA A receptor shows a ligand-gated receptor super-family

Amino-acid sequences derived from complementary DNAs encoding the alpha- and beta-subunits of the GABA/benzodiazepine receptor from bovine brain show homology with other ligand-gated receptor subunits, suggesting that there is a super-family of ion-channel-containing receptors. Co-expression of the in vitro-generated alpha-subunit and beta-subunit RNAs in Xenopus oocytes produces a functional receptor and ion channel with the pharmacological properties characteristic of the GABAA receptor.     

Intracellular calcium stores regulate activity-dependent neuropeptide release from dendrites

Information in neurons flows from synapses, through the dendrites and cell body (soma), and, finally, along the axon as spikes of electrical activity that will ultimately release neurotransmitters from the nerve terminals. However, the dendrites of many neurons also have a secretory role, transmitting information back to afferent nerve terminals. In some central nervous system neurons, spikes that originate at the soma can travel along dendrites as well as axons, and may thus elicit secretion from both compartments. Here, we show that in hypothalamic oxytocin neurons, agents that mobilize intracellular Ca(2+) induce oxytocin release from dendrites without increasing the electrical activity of the cell body, and without inducing secretion from the nerve terminals. Conversely, electrical activity in the cell bodies can cause the secretion of oxytocin from nerve terminals with little or no release from the dendrites. Finally, mobilization of intracellular Ca(2+) can also prime the releasable pool of oxytocin in the dendrites. This priming action makes dendritic oxytocin available for release in response to subsequent spike activity. Priming persists for a prolonged period, changing the nature of interactions between oxytocin neurons and their neighbours.     

Intracellular free calcium rise triggers nuclear envelope breakdown in the sea urchin embryo

Cytosolic free calcium has recently been implicated in the regulation of mitosis in plant and animal cells. We have previously found correlations between increases in the levels of intracellular free calcium [Ca2+]i and visible transitions of structure at nuclear envelope breakdown (NEBD) and the onset of anaphase during mitosis in sea urchin embryos and tissue culture cells. To go beyond correlations it is necessary to manipulate [Ca2+]i, and in sea urchin embryos this requires the injection of calcium-chelator buffer solutions as the changes in free calcium in the cell cycle are dependent on intracellular stores. We report here that blocking the increase in [Ca2+]i which just precedes NEBD prevents this from taking place and halts mitosis. Subsequent injections which momentarily increase [Ca2+]i, or a natural recovery of the higher calcium levels, result in NEBD and the successful continuation of mitosis. Similarly, artificially increasing calcium by early injections results in early NEBD. We conclude that the increase in [Ca2+]i preceding NEBD is an essential regulatory step required for entry into mitosis.     

Intracellular calcium dependence of transmitter release rates at a fast central synapse

Calcium-triggered fusion of synaptic vesicles and neurotransmitter release are fundamental signalling steps in the central nervous system. It is generally assumed that fast transmitter release is triggered by elevations in intracellular calcium concentration ([Ca2+]i) to at least 100 microM near the sites of vesicle fusion. For synapses in the central nervous system, however, there are no experimental estimates of this local [Ca2+]i signal. Here we show, by using calcium ion uncaging in the large synaptic terminals of the calyx of Held, that step-like elevations to only 10 microM [Ca2+]i induce fast transmitter release, which depletes around 80% of a pool of available vesicles in less than 3 ms. Kinetic analysis of transmitter release rates after [Ca2+]i steps revealed the rate constants for calcium binding and vesicle fusion. These show that transient (around 0.5 ms) local elevations of [Ca2+]i to peak values as low as 25 microM can account for transmitter release during single presynaptic action potentials. The calcium sensors for vesicle fusion are far from saturation at normal release probability. This non-saturation, and the high intracellular calcium cooperativity in triggering vesicle fusion, make fast synaptic transmission very sensitive to modulation by changes in local [Ca2+]i.     

钙离子的浓度对X80钢腐蚀行为的影响

采用失重法、电化学测试、扫描电镜、X射线衍射等方法研究了钙离子浓度对X80钢在哈密土壤模拟溶液中的腐蚀行为影响.在60 d浸泡期内,X80钢在不同钙离子浓度模拟溶液中的腐蚀形态均为全面腐蚀,腐蚀产物都为β-FeOOH;X80钢在模拟溶液中的腐蚀速率随钙离子浓度的降低而呈逐渐增大的趋势.在180 d浸泡期内,在钙离子浓度为63.5 mmol·L-1的模拟溶液中,钙盐随时间的增加在X80钢基体表面不断结晶析出;钙盐层有效阻碍了溶解氧的迁移,并促进其覆盖区域下形成氧浓差电池,最终导致基体表面点蚀的萌生.同时,在内层腐蚀产物表面连续析出的钙盐层的致密性也随时间不断得到改善,在一定程度上起到了抑制氯离子和溶解氧对基体的侵蚀作用,X80钢的全面腐蚀逐渐减缓.     

Coupling of agonist binding to channel gating in the GABA(A) receptor

Neurotransmitters such as acetylcholine and GABA (gamma-aminobutyric acid) mediate rapid synaptic transmission by activating receptors belonging to the gene superfamily of ligand-gated ion channels (LGICs). These channels are pentameric proteins that function as signal transducers, converting chemical messages into electrical signals. Neurotransmitters activate LGICs by interacting with a ligand-binding site, triggering a conformational change in the protein that results in the opening of an ion channel. This process, which is known as 'gating', occurs rapidly and reversibly, but the molecular rearrangements involved are not well understood. Here we show that optimal gating in the GABA(A) receptor, a member of the LGIC superfamily, is dependent on electrostatic interactions between the negatively charged Asp 57 and Asp 149 residues in extracellular loops 2 and 7, and the positively charged Lys 279 residue in the transmembrane 2-3 linker region of the alpha1-subunit. During gating, Asp 149 and Lys 279 seem to move closer to one another, providing a potential mechanism for the coupling of ligand binding to opening of the ion channel.     

Activation of mitochondrial oxidative metabolism by calcium ions in Limulus ventral photoreceptor

Cells regulate their metabolic energy production to meet the requirements of their energy consuming activities. For most animal cells the prime site of energy production, in the form of ATP, is the mitochondrion. Extensive in vitro studies of isolated mitochondria have provided detailed information about the specific biochemical reactions involved in energy production. At present there is a debate about whether respiration in excitable cells is controlled by the availability of ADP to the mitochondrion and/or by calcium ions. Using the large ventral photoreceptor of the horseshoe crab (Limulus polyphemus) we describe a method for measuring the transient increase in the mitochondrial O2 consumption (delta QO2) following a flash of light of a single photoreceptor. We then show that this delta QO2 results in part from a rise in the intracellular concentration of calcium (Cai).     

包载铜离子无定形磷酸钙的制备及表征

采用共沉淀法并加入有机双膦酸作为改性剂,制备出了能够包载铜离子的稳定的无定形磷酸钙.通过傅立叶红外吸收光谱分析(FTIR),X射线衍射(XRD),热重分析(TGA)对材料的化学结构和组成进行了表征.FTIR和TGA表明有机双膦酸能够稳定的接到磷酸钙上使磷酸钙的结晶度降低.XRD表明材料能够稳定的包载铜离子并具有无定形结构.     

Polycystin-L is a calcium-regulated cation channel permeable to calcium ions.

Polycystic kidney diseases are genetic disorders in which the renal parenchyma is progressively replaced by fluid-filled cysts. Two members of the polycystin family (polycystin-1 and -2) are mutated in autosomal dominant polycystic kidney disease (ADPKD), and polycystin-L is deleted in mice with renal and retinal defects. Polycystins are membrane proteins that share significant sequence homology, especially polycystin-2 and -L (50% identity and 71% similarity). The functions of the polycystins remain unknown. Here we show that polycystin-L is a calcium-modulated nonselective cation channel that is permeable to sodium, potassium and calcium ions. Patch-clamp experiments revealed single-channel activity with a unitary conductance of 137 pS. Channel activity was substantially increased when either the extracellular or intracellular calcium-ion concentration was raised, indicating that polycystin-L may act as a transducer of calcium-mediated signalling in vivo. Its large single-channel conductance and regulation by calcium ions distinguish it from other structurally related cation channels.     

Highly cooperative feedback control of retinal rod guanylate cyclase by calcium ions

Visual excitation in retinal rod cells is mediated by a cascade that leads to the amplified hydrolysis of cyclic GMP (cGMP) and the consequent closure of cGMP-activated cation-specific channels in the plasma membrane. Recovery of the dark state requires the resynthesis of cGMP, which is catalysed by guanylate cyclase, an axoneme-associated enzyme. The lowering of the cytosolic calcium concentration (Cai) following illumination is thought to be important in stimulating cyclase activity. This hypothesis is supported by the finding that the cGMP content of rod outer segments increases several-fold when Cai is lowered to less than 10 nM. It is evident that cGMP and Cai levels are reciprocally controlled by negative feedback. Guanylate cyclase from toad ROS is strongly stimulated when the calcium level is lowered from 10 microM to 10 nM, but only if they are excited by light. We show here that the guanylate cyclase activity of unilluminated bovine rod outer segments increases markedly (5 to 20-fold) when the calcium level is lowered from 200 nM to 50 nM. This steep dependence of guanylate cyclase activity on the calcium level in the physiological range has a Hill coefficient of 3.9. Stimulation at low calcium levels is mediated by a protein that can be released from the outer segment membranes by washing with a low salt buffer. Calcium sensitivity is partially restored by adding the soluble extract back to the washed membranes. The highly cooperative activation of guanylate cyclase by the light-induced lowering of Cai is likely to be a key event in restoring the dark current after excitation.     

GABA转运蛋白研究进展

GABA在逆境条件下能影响植物的生长,提高植物的耐性对农业生产具有重大的意义.对于GABA在植物逆境中的积累机制及代谢功能现已经有了大量的研究,GABA在植物体内积累与代谢机制与质膜上的转运蛋白密切相关.本文就GABA转运蛋白的结构与功能,以及在动植物中的发现,和自身的磷酸化与去磷酸化机制进行了归纳与总结.