Retinal light adaptation--evidence for a feedback mechanism

Light adaptation is the adjustment of retinal response properties to variations in ambient illumination. It enables the encoding of visual information over a millionfold intensity range, from moonlight to broad daylight, despite the relatively small dynamic range of response of visual neurones. We have studied the effects of light adaptation on the dynamics and sensitivity of visual responses of neurones in the turtle retina, by measuring the responses of horizontal cells in the retina to light which was modulated with a sinusoidal time course around various mean levels. As a quantitative measure of the transduction from light to neural signals, we calculated the gain of response at each frequency. Gain is defined as the amplitude of the modulated response component divided by the amplitude of light modulation. We report here that the gain (mV photon-1) at low temporal frequencies decreased as the mean light level increased. Over a 2 log-unit range of mean light levels, low-frequency gain was inversely proportional to the mean light level, as in Weber's law. However, at high temporal frequencies, the gain was almost independent of mean light level. Our results are reminiscent of Kelly's results on human temporal-frequency sensitivity in various states of light adaptation. We found that a family of horizontal-cell temporal frequency responses, measured at various mean light levels, could be accounted for by a negative feedback model in which the feedback strength is proportional to mean light level.     

A presynaptic action of glutamate at the cone output synapse

Neurotransmitter release from many central nervous system synapses is regulated by 'autoreceptors' at the synaptic terminal, which bind the released transmitter and alter release accordingly. The photoreceptors of lower vertebrates are thought to use glutamate as a neurotransmitter. Glutamate conveys the visual signal to postsynaptic bipolar and horizontal cells, but has been reported not to act on the photoreceptors themselves. We show here that glutamate evokes a current, carried largely by chloride ions, in cones isolated from the tiger salamander retina. This response is localized to the synaptic terminal of the cone. Removing external sodium blocks this action of glutamate. These results suggest the existence of a positive feedback loop at the cone output synapse: over most of the light-response range, glutamate released by depolarization of the cone will cause further depolarization, increasing the gain of phototransduction. Glutamate released from rods may also polarize cones, modulating the gain of the cone output synapse. This system is surprisingly different from the autoreceptor systems for most other transmitters, which act in a negative feedback way.     

Role of visual pigment properties in rod and cone phototransduction

Retinal rods and cones share a phototransduction pathway involving cyclic GMP. Cones are typically 100 times less photosensitive than rods and their response kinetics are several times faster, but the underlying mechanisms remain largely unknown. Almost all proteins involved in phototransduction have distinct rod and cone variants. Differences in properties between rod and cone pigments have been described, such as a 10-fold shorter lifetime of the meta-II state (active conformation) of cone pigment and its higher rate of spontaneous isomerization, but their contributions to the functional differences between rods and cones remain speculative. We have addressed this question by expressing human or salamander red cone pigment in Xenopus rods, and human rod pigment in Xenopus cones. Here we show that rod and cone pigments when present in the same cell produce light responses with identical amplification and kinetics, thereby ruling out any difference in their signalling properties. However, red cone pigment isomerizes spontaneously 10,000 times more frequently than rod pigment. This high spontaneous activity adapts the native cones even in darkness, making them less sensitive and kinetically faster than rods. Nevertheless, additional factors are probably involved in these differences.     

Cyclic GMP-sensitive conductance in outer segment membrane of catfish cones

A cyclic GMP-sensitive conductance has recently been observed with patch-clamp recording in excised inside-out patches of plasma membrane from frog and toad rod outer segments. This conductance has properties suggesting that it is probably the light-sensitive conductance involved in visual transduction. We now report a similar conductance in the outer segment membrane of catfish cones. Cyclic GMP showed positive cooperativity in opening this conductance, with a Hill coefficient of 1.6-3.0 and a half-saturating cGMP concentration of 35-70 microM. Cyclic AMP at 1 mM, or changing Ca concentration (in the presence of Mg), had little effect on the conductance. In physiological solutions the cGMP-induced current had a reversal potential near +10 mV; the current amplitude increased roughly exponentially with membrane potential in both depolarizing and hyperpolarizing directions. Our results suggest that cGMP is also the internal transmitter for phototransduction in cones.     

Restoration of vision after transplantation of photoreceptors

Cell transplantation is a potential strategy for treating blindness caused by the loss of photoreceptors. Although transplanted rod-precursor cells are able to migrate into the adult retina and differentiate to acquire the specialized morphological features of mature photoreceptor cells, the fundamental question remains whether transplantation of photoreceptor cells can actually improve vision. Here we provide evidence of functional rod-mediated vision after photoreceptor transplantation in adult Gnat1?/? mice, which lack rod function and are a model of congenital stationary night blindness. We show that transplanted rod precursors form classic triad synaptic connections with second-order bipolar and horizontal cells in the recipient retina. The newly integrated photoreceptor cells are light-responsive with dim-flash kinetics similar to adult wild-type photoreceptors. By using intrinsic imaging under scotopic conditions we demonstrate that visual signals generated by transplanted rods are projected to higher visual areas, including V1. Moreover, these cells are capable of driving optokinetic head tracking and visually guided behaviour in the Gnat1?/? mouse under scotopic conditions. Together, these results demonstrate the feasibility of photoreceptor transplantation as a therapeutic strategy for restoring vision after retinal degeneration.     

Cyclic GMP is involved in the excitation of invertebrate photoreceptors

The hyperpolarizing receptor potential in vertebrate rod photoreceptors appears to be mediated by the second messenger, cyclic GMP. Injection of cGMP into rods or application of cGMP to inside-out membrane patches activates a conductance resembling that produced by light. Light produces a rapid reduction of cGMP in living rods, leading to closure of sodium channels and membrane hyperpolarization. In most invertebrate photoreceptors the response to light is depolarizing. We have investigated whether cGMP is involved in controlling the increase in sodium conductance that underlies this depolarization. We show here that injection of cGMP into Limulus photoreceptors produces a depolarization that mimics the receptor potential. We also show that the cGMP concentration of the squid retina increases rapidly during exposure to light. These results support the hypothesis that cGMP mediates the light-induced depolarization in invertebrate photoreceptors and suggests that vertebrate and invertebrate phototransduction may be more similar than previously thought.     

Effect of bleached rhodopsin on signal amplification in rod visual receptors

Bleaching of rhodopsin markedly desensitizes the vertebrate visual system during a subsequent period of dark adaptation. Previous studies have indicated an origin of bleaching desensitization in the visual pigment itself, but have not identified the mechanism of action. A candidate for the site at which densensitization is initially expressed is the activation of transducin (formation of T*) on the rod disk membranes; this reaction directly involves rhodopsin in its photoactivated (R*) form and mediates initial amplification of the visual signal (reviewed in refs 7-9). We have analysed the effect of bleaching on the sensitivity of a flash-induced light-scattering signal known to monitor the disk-based amplifier, and which has been established as specifically monitoring transducin activation. We have recorded this signal from functioning retinal rods in situ ('ATR' signal) and find that bleaches inducing a pronounced, sustained loss in rod electrophysiological sensitivity do not alter the sensitivity of the ATR response after correction for reduced quantum catch. Our results indicate that the biochemical gain of the R*----T* transduction stage remains unchanged in the presence of bleached pigment and implicate a subsequent reaction as the first to show a sustained, bleaching-dependent gain reduction.     

偏磨抽油杆接箍力学性能及承载能力分析——以塔河油田抽油杆接箍断脱为例

针对塔河油田抽油杆接箍在采油生产过程中,易发生磨损现象,而导致抽油杆接箍断脱失效的问题。以直径为25mm抽油杆,基体材料为35CrMo接箍为研究对象,考虑接箍壁厚受磨损程度的影响,应用有限元方法,开展抽油杆接箍在拉压载荷作用下受力分析和承载范围分析。通过拉压试验测试和数值模拟结果对比分析并验证数值方法可靠性基础上,开展了拉压载荷在20KN-250KN下不同磨损量抽油杆接箍的力学性能分析。研究表明,等效应力和接触应力均随着载荷增大而增大,无磨损接箍在受力范围内均能满足应用要求。但由于接箍出现偏磨,其等效应力与接触应力分布对称性变差,且磨损严重时,其承载能力严重下降,断脱风险加大。     

cGMP-dependent cation channel of retinal rod outer segments

Light-modulated cytoplasmic cGMP simultaneously controls plasma membrane Na+ conductance in visual excitation and Ca2+ entry into rods by direct interaction with the cation channel. Cytoplasmic Ca2+ in turn may set operating points and contribute to the dynamics of several enzymes that regulate cGMP levels in the dark, recovery from excitation and receptor adaptation or down regulation. Similar channels may couple electrical activity to internal nucleotide metabolism in other tissues. We here report the identification, partial purification and behaviour after reconstitution of a protein of relative molecular mass 39,000 (Mr 39K) present in both disk and plasma membranes from bovine rod outer segments that mediates these cGMP-dependent cation fluxes. Its cGMP agonist specificity, kinetic cooperativity, ionic selectivity, membrane density and other features closely match the properties of the visual cGMP-dependent conductance inferred from electrophysiological measurements.     

神经元轴突电刺激响应的有限元数值模拟

为了研究神经元轴突的微观生理电传导特性,建立了神经元轴突的三维有限元模型,通过数值计算模拟神经元轴突对电刺激的动态响应.建立海马神经元轴突的三维几何模型并指定其生物物理参数,根据Hodgkin-Huxley方程和Maxwell方程,建立偏微分方程组,对神经元轴突有限元模型施加不同持续时间和不同脉冲幅度的电流脉冲并求解,获得神经元轴突的三维电势分布和动作电位曲线.数值模拟结果显示,该神经元轴突的静息电位约为-65 mV;对模型施加持续时间为2 ms,强度0.01 A/m2的电流脉冲刺激未产生动作电位,施以(2 ms、0.2 A/m2),(20ms、0.01 A/m2),(20ms、0.2 A/m2)脉冲刺激均产生动作电位,峰值分别出现在0.012 s、0.017 s和0.012 s,动作电位幅度约为100mV,持续时间约为2 ms.神经元轴突电刺激响应的有限元模拟结果与实验结果吻合,表明所建立的神经元轴突有限元模型及数值模拟方法合理、可靠,为深入研究神经电生理特性提供了基础模型和仿真分析方法.     

离散杆战斗部杆条飞行性能模拟试验研究

通过离散杆战斗部靶场静爆试验,对离散杆式破片的飞行性能进行了试验研究。试验结果表明：采用波形控制器可以较好地控制战斗部爆炸后杆形破片的飞散方向,以及保证杆务的完整率;杆务的飞行姿态与放置角有密切关系,主要通过杆条旋转速度表现出来。     

Novel preparation for studying excitation-secretion coupling in the isolated single cell

Cellular mechanisms of secretion in exocrine and endocrine glands are technically difficult to study. We present here a model which offers fundamental advantages for studying excitation-secretion coupling at the level of the isolated single cell. The salivary gland of the leech Haementeria ghilianii possesses a unique combination of unusual properties which greatly facilitate research in this area. Its cells are exceptionally large (up to 1 mm in diameter), clearly visible and easy to penetrate with microelectrodes. They do not form a homogeneous population but consist of five distinct histochemical types, secreting a number of identified products such as the fibrinolytic enzyme hementin (for which there is a sensitive assay). The cells generate overshooting calcium-dependent action potentials up to 90 mV in amplitude and with a duration of 200-1,000 ms. One of their most useful and unusual features is a lack of electrical coupling which means that individual cells can be studied in an intact gland without interference from neighbouring cells.     

Induction by cyclic GMP of cationic conductance in plasma membrane of retinal rod outer segment

Vertebrate rod photoreceptors hyperpolarize when illuminated, due to the closing of cation-selective channels in the plasma membrane. The mechanism controlling the opening and closing of these channels is still unclear, however. Both 3',5'-cyclic GMP and Ca2+ ions have been proposed as intracellular messengers for coupling the light activation of the photopigment rhodopsin to channel activity and thus modulating light-sensitive conductance. We have now studied the effects of possible conductance modulators on excised 'inside-out' patches from the plasma membrane of the rod outer segment (ROS), and have found that cyclic GMP acting from the inner side of the membrane markedly increases the cationic conductance of such patches (EC50 30 microM cyclic GMP) in a reversible manner, while Ca2+ is ineffective. The cyclic GMP-induced conductance increase occurs in the absence of nucleoside triphosphates and, hence, is not mediated by protein phosphorylation, but seems rather to result from a direct action of cyclic GMP on the membrane. The effect of cyclic GMP is highly specific; cyclic AMP and 2',3'-cyclic GMP are completely ineffective when applied in millimolar concentrations. We were unable to recognize discrete current steps that might represent single-channel openings and closings modulated by cyclic GMP. Analysis of membrane current noise shows the elementary event to be 3 fA with 110 mM Na+ on both sides of the membrane at a membrane potential of -30 mV. If the initial event is assumed to be the closure of a single cyclic GMP-sensitive channel, this value corresponds to a single-channel conductance of 100 fS. It seems probable that the cyclic GMP-sensitive conductance is responsible for the generation of the rod photoresponse in vivo.     

Preferential electrical coupling regulates neocortical lineage-dependent microcircuit assembly

Radial glial cells are the primary neural progenitor cells in the developing neocortex. Consecutive asymmetric divisions of individual radial glial progenitor cells produce a number of sister excitatory neurons that migrate along the elongated radial glial fibre, resulting in the formation of ontogenetic columns. Moreover, sister excitatory neurons in ontogenetic columns preferentially develop specific chemical synapses with each other rather than with nearby non-siblings. Although these findings provide crucial insight into the emergence of functional columns in the neocortex, little is known about the basis of this lineage-dependent assembly of excitatory neuron microcircuits at single-cell resolution. Here we show that transient electrical coupling between radially aligned sister excitatory neurons regulates the subsequent formation of specific chemical synapses in the neocortex. Multiple-electrode whole-cell recordings showed that sister excitatory neurons preferentially form strong electrical coupling with each other rather than with adjacent non-sister excitatory neurons during early postnatal stages. This preferential coupling allows selective electrical communication between sister excitatory neurons, promoting their action potential generation and synchronous firing. Interestingly, although this electrical communication largely disappears before the appearance of chemical synapses, blockade of the electrical communication impairs the subsequent formation of specific chemical synapses between sister excitatory neurons in ontogenetic columns. These results suggest a strong link between lineage-dependent transient electrical coupling and the assembly of precise excitatory neuron microcircuits in the neocortex.     

三维油藏流动电位数值模拟及油水前缘预测

根据流动电位产生的基本原理,建立三维油水两相流动条件下电场和渗流场的耦合数值模型并采用有限差分方法进行求解;以四分之一"五点法"井网为对象,模拟计算一注一采生产过程中井底的电位变化。结果表明:油水前缘的位置会出现电位的峰值,该峰值随注入水向生产井一侧移动,生产井底的电位会连续上升,当油水前缘距离生产井75 m时开始剧烈变化,因此对该效应的监测及分析能够实现油水前缘的远距离预测;注入水沿高渗透层的快速前进会引起电位异常,使高渗透层比低渗透层要高出5~15 mV,通过对该信号的强弱和随时间的变化规律的分析能够识别地层非均质性和注入水的不均匀推进,该方法具有成为新型动态监测手段的潜力。     

内插中心斜杆换热管的换热性能

在热传递技术的被动改进中,在管中使用插入物是当今一种非常普遍并且具有很大实际作用的改进技术。内插中心斜杆换热管可以使管内实现类似于优化流场的多纵向涡旋流,使换热管在流动阻力增幅不大的情况下换热性能得到有效提升。利用数值模拟方法对内插中心斜杆的换热管进行研究,探求斜杆数目、节距、直径等参数的变化对换热、阻力特性的影响。结果发现:内插中心斜杆换热管的换热性能远优于光管的综合换热性能;内插中心斜杆换热管的努塞尔数随着斜杆数量的增多而在一定范围提高,压降随着斜杆数量的增多而增大,斜杆数量为3时,内插中心斜杆换热管的综合换热性能较好;努塞尔数和压降随着节距的增大而减小,斜杆节距为20 mm时,内插中心斜杆换热管的综合换热性能较好;努塞尔数和压降随着斜杆直径的增大而增加,斜杆直径为2.0 mm时,内插中心斜杆换热管的综合换热性能较好。     

引进及国产大截面塑料注射模脱模顶杆的失效行为

研究了引进日本及国产大截面塑料注射模脱模顶杆在相近锁模力下的失效行为.分析结果表明,日本顶杆在注塑50万次后以磨损的形式失效,国产顶杆在注塑10万次后以弯曲断裂的形式失效.前者比后者具有高的使用寿命,这是因为日本顶杆选用的是含铬的结构钢,并采取了调质处理后再表面淬火的热处理工艺,使之获得了表层高硬度及心部综合机械性能好的特性,从而阻止了早期失效行为的出现.     

Melanopsin and rod-cone photoreceptive systems account for all major accessory visual functions in mice

In the mammalian retina, besides the conventional rod-cone system, a melanopsin-associated photoreceptive system exists that conveys photic information for accessory visual functions such as pupillary light reflex and circadian photo-entrainment. On ablation of the melanopsin gene, retinal ganglion cells that normally express melanopsin are no longer intrinsically photosensitive. Furthermore, pupil reflex, light-induced phase delays of the circadian clock and period lengthening of the circadian rhythm in constant light are all partially impaired. Here, we investigated whether additional photoreceptive systems participate in these responses. Using mice lacking rods and cones, we measured the action spectrum for phase-shifting the circadian rhythm of locomotor behaviour. This spectrum matches that for the pupillary light reflex in mice of the same genotype, and that for the intrinsic photosensitivity of the melanopsin-expressing retinal ganglion cells. We have also generated mice lacking melanopsin coupled with disabled rod and cone phototransduction mechanisms. These animals have an intact retina but fail to show any significant pupil reflex, to entrain to light/dark cycles, and to show any masking response to light. Thus, the rod-cone and melanopsin systems together seem to provide all of the photic input for these accessory visual functions.     

Metarhodopsin I/metarhodopsin II transition triggers light-induced change in calcium binding at rod disk membranes

The hypothesis of Yoshikami and Hagins that calcium ions act as diffusible transmitter molecules between the photochemistry of rhodopsin and the subsequent electrical events at the outer plasma membrane of rods initiated many investigations on light-stimulated calcium release in vertebrate photoreceptor cells (see refs 2, 3). Although it not seems firmly established that light has some effect on the redistribution of calcium in various disk preparations, reconstituted systems and intact rod outer segments, the physiological significance remained unclear. We previously reported a rapid, light-triggered calcium release from binding sites at the disk membrane in the presence of calcium ionophore A23187 (refs 3, 8). However, there is no evidence for rapid calcium release into the cytosol in the absence of ionophore. On fragmentation of intact rod outer segments, calcium release due to a light-requlated change of calcium binding appeared almost completely abolished. We describe here experiments with sonicated rod outer segments in which the previously observed loss of the calcium release capacity has been prevented. Calcium release in sonicated disks in the presence of A23187 kinetically follows the metarhodopsin I/metarhodopsin II transition (tau 1/2 = 10 ms, activation energy EA = 34 kcal mol-1), suggesting that calcium release is triggered by this photochemical transition.     

Cyclic GMP can increase rod outer-segment light-sensitive current 10-fold without delay of excitation

To test the hypothesis that cyclic GMP is the internal messenger coupling rhodopsin activation to membrane excitation in vertebrate rod photoreceptors, we used a novel technique combining measurement of membrane currents of isolated salamander rods with a suction electrode and the introduction of cyclic GMP through a whole-cell recording patch pipette. Rupture of an attached patch was followed by a rapid (approximately 10 s), approximately 10-fold increase in outer-segment membrane current, all of which was light-sensitive. There was little change in the rising phase of the response to a saturating flash, but the duration of the saturated phase of the response increased approximately 10-fold. The effects reversed completely within 3-4 min after withdrawal of the cyclic GMP-containing patch pipette. A formal kinetic analysis shows that the first two observations are inconsistent with the postulate that cyclic GMP opens the light-sensitive conductance by simple binding to channels, unless free cyclic GMP in the outer segment is assumed to be much lower than published estimates, and most of the outer-segment cyclic GMP is bound and inexchangeable on the timescale of 200 ms. Furthermore, our results suggest that rod cyclic GMP is not involved solely in keeping the light-sensitive conductance open, but may also affect the activity of the phosphodiesterase that mediates cyclic GMP hydrolysis.