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.     

Spectral sensitivity of a novel photoreceptive system mediating entrainment of mammalian circadian rhythms

Environmental light cycles are the dominant synchronizers of circadian rhythms in the field, and artificial light cycles and pulses are the major tools used in the laboratory to analyse properties of circadian systems. It is therefore surprising that few studies have analysed the physical parameters of light stimuli that affect circadian rhythms. There have previously been no spectral sensitivity measurements for phase shifting the circadian rhythms of mammals and only two preliminary reports on the wavelength dependence of this response exist. Using the magnitude of phase shift caused by a single 15-min pulse of monochromatic light given 6 h after activity onset, we have now characterized the spectral sensitivity of the photoreceptors responsible for phase shifting the locomotor rhythm of the hamster (Mesocricetus auratus). The sensitivity curve for this response has a maximum near 500 nm and is similar to the absorption spectrum for rhodopsin. Although the spectral sensitivity is consistent with a rhodopsin-based photopigment, two features of the photoreceptive system that mediates entrainment are unusual: the threshold of the response is high, especially for a predominantly rod retina like that of the hamster, and the reciprocal relationship between intensity and duration holds for extremely long durations (up to 45 min). These results suggest that the photoreceptive system mediating entrainment is markedly different from that involved in visual image formation.     

蟾蜍松果体的细胞学观察及其冬眠期与非冬眠期形态学的比较研究

为了研究蟾蜍松果体的形态特征,并了解松果体在生物节律中所起的作用,运用常规的光镜样品制备技术,对松果体的细胞形态及其在冬眠期、非冬眠期的变化进行了观察.研究结果显示,松果体的额器及脑上体都具有与视网膜感光细胞相似的细胞,称之为类光感细胞,因而它们具有感受光的能力;同时,根据细胞结构的观察以及两者所处位置的不同,认为额器具有比脑上体更强的感光性能.通过冬眠期与非冬眠期的细胞形态学比较,可以看到额器及脑上体的类光感细胞外节均有较明显的变化.冬眠期可见额器的类光感细胞外节排列规则,但数量减少,而脑上体的类光感细胞外节排列明显不规则,内有很多小泡,呈退化状态.额器及脑上体支持细胞的细胞质和类光感细胞线粒体的形态在冬眠扨亦有一定的变化.实验结果表明,额器及脑上体感光性能在冬眠期与非冬眠期是有差异的,脑上体在冬眠斯进入退化状态。     

Melanopsin-expressing ganglion cells in primate retina signal colour and irradiance and project to the LGN

Human vision starts with the activation of rod photoreceptors in dim light and short (S)-, medium (M)-, and long (L)- wavelength-sensitive cone photoreceptors in daylight. Recently a parallel, non-rod, non-cone photoreceptive pathway, arising from a population of retinal ganglion cells, was discovered in nocturnal rodents. These ganglion cells express the putative photopigment melanopsin and by signalling gross changes in light intensity serve the subconscious, 'non-image-forming' functions of circadian photoentrainment and pupil constriction. Here we show an anatomically distinct population of 'giant', melanopsin-expressing ganglion cells in the primate retina that, in addition to being intrinsically photosensitive, are strongly activated by rods and cones, and display a rare, S-Off, (L + M)-On type of colour-opponent receptive field. The intrinsic, rod and (L + M) cone-derived light responses combine in these giant cells to signal irradiance over the full dynamic range of human vision. In accordance with cone-based colour opponency, the giant cells project to the lateral geniculate nucleus, the thalamic relay to primary visual cortex. Thus, in the diurnal trichromatic primate, 'non-image-forming' and conventional 'image-forming' retinal pathways are merged, and the melanopsin-based signal might contribute to conscious visual perception.     

A new role for cryptochrome in a Drosophila circadian oscillator

Cryptochromes are flavin/pterin-containing proteins that are involved in circadian clock function in Drosophila and mice. In mice, the cryptochromes Cry1 and Cry2 are integral components of the circadian oscillator within the brain and contribute to circadian photoreception in the retina. In Drosophila, cryptochrome (CRY) acts as a photoreceptor that mediates light input to circadian oscillators in both brain and peripheral tissue. A Drosophila cry mutant, cryb, leaves circadian oscillator function intact in central circadian pacemaker neurons but renders peripheral circadian oscillators largely arrhythmic. Although this arrhythmicity could be caused by a loss of light entrainment, it is also consistent with a role for CRY in the oscillator. A peripheral oscillator drives circadian olfactory responses in Drosophila antennae. Here we show that CRY contributes to oscillator function and physiological output rhythms in the antenna during and after entrainment to light-dark cycles and after photic input is eliminated by entraining flies to temperature cycles. These results demonstrate a photoreceptor-independent role for CRY in the periphery and imply fundamental differences between central and peripheral oscillator mechanisms in Drosophila.     

金鱼视觉系统内中、高分子量神经丝免疫组织化学定位

RT97是一种特异性识别中、高分子量神经丝蛋白抗原簇的抗体.采用组织化学技术对金鱼(Carassiusauratus)视觉系统内RT97识别神经丝的表达分布进行了观察.结果表明,RT97在金鱼视网膜、视神经和视顶盖中均有阳性反应:①在视网膜内视纤维层、内网层、内核层的水平细胞、光感受器的外节有RT97阳性反应;②在视乳突部和视神经有少量生长纤维为RT97阳性;③在视顶盖中RT97阳性反应集中在顶盖的边缘层(SM),同时观察到有RT97阳性纤维从中央纵突区(TL)通过顶盖腹面延伸至顶盖边缘区.实验结果提示,RT97不仅可作为金鱼视觉系统视觉生长纤维的一种标记蛋白,同时在视顶盖由RT97识别的蛋白也可能是一种吸引视觉纤维生长的靶蛋白.     

匈牙利田鼠视交叉上核的研究

哺乳动物下丘脑的视交叉上核控制着多种生理节奏的发生,田鼠是一种营地下生活的独特的啮齿动物,用免疫组织化学,辣根过氧化物酶法和Ｎｉｓｓｌｅ染色法做了其ＳＣＮ形态上的一些研究。     

A cyclic GMP-activated channel in dissociated cells of the chick pineal gland.

Phototransduction in the vertebrate retina is dependent in part on a cyclic GMP-activated ionic channel in the plasma membrane of rods and cones. But other vertebrate cells are also photosensitive. Cells of the chick pineal gland have a photosensitive circadian rhythm in melatonin secretion that persists in dissociated cell culture. Exposure to light causes inhibition of melatonin secretion, and entrainment of the intrinsic circadian oscillator. Chick pinealocytes express several 'retinal' proteins, including arrestin, transducin and a protein similar to the visual pigment rhodopsin. Pinealocytes of lower vertebrates display hyperpolarizing responses to brief pulses of light. Thus it is possible that some of the mechanisms of phototransduction are similar in retinal and pineal photoreceptors. We report here the first recordings of cyclic GMP-activated channels in an extraretinal photoreceptor. Application of GMP, but not cyclic AMP, to excised inside-out patches caused activation of a 15-25 pS cationic channel. These channels may be essential for phototransduction in the chick pineal gland.     

动物感光器中的G蛋白及其偶联的光信号转导

G蛋白偶联的信号转导系统是细胞跨膜信号转导的重要途径，在动物的光感觉生理中，G蛋白在信号的转导和放大过程中起了重要的作用，有关这方面的研究已是动物光感觉研究中的一个热点，作者介绍了国内外这方面的研究进展，以及研究的方法，并对动物感光器中G蛋白的类型、性质和功能作了简要的概括。     

刀额新对虾复眼的外部形态结构及光、暗适应对其超微结构的影响

应用电镜技术观察了刀额新对虾复眼的形态结构以及光、暗适应对其超微结构的影响．结果表明，刀额新对虾复眼外形呈圆球形，约由93116个小眼组成，组成复眼的小眼面均为正方形．每个小眼由折光系统和感光系统组成．光、暗适应时折光系统的结构基本相同，但感光系统之间有较大的差异．暗适应时，组成感杆束的微纤毛排列整齐，膜下储泡囊数量多，体积大，色素颗粒仅分布在细胞体远端；而光适应时，组成感杆束的微纤毛排列凌乱，并有部分溶解，多囊体多，各个层面上有板膜体的出现，色素颗粒分布于整个细胞．     

Light-induced suppression of endogenous circadian amplitude in humans

Winfree reported 20 years ago the intriguing finding that a light stimulus of a critical strength applied at a critical circadian phase could essentially stop the circadian clock in Drosophila pseudo-obscura by resetting the circadian oscillator close to its singularity (a phaseless position at which the amplitude of circadian oscillation is zero). Since then, similar observations of attenuated circadian amplitude in response to critical stimuli have been limited to unicells, insects and plants. Our recent demonstration that the phase of the human circadian pacemaker could be inverted using an unconventional three-cycle stimulus led us to investigate whether critically timed exposure to a more moderate stimulus could drive that oscillator towards its singularity. Here we report that exposure of humans to fewer cycles of bright light, centred around the time at which the human circadian pacemaker is most sensitive to light-induced phase shifts, can markedly attenuate endogenous circadian amplitude. In some cases this results in an apparent loss of rhythmicity, as expected to occur in the region of singularity.     

Addition of human melanopsin renders mammalian cells photoresponsive

A small number of mammalian retinal ganglion cells act as photoreceptors for regulating certain non-image forming photoresponses. These intrinsically photosensitive retinal ganglion cells express the putative photopigment melanopsin. Ablation of the melanopsin gene renders these cells insensitive to light; however, the precise role of melanopsin in supporting cellular photosensitivity is unconfirmed. Here we show that heterologous expression of human melanopsin in a mouse paraneuronal cell line (Neuro-2a) is sufficient to render these cells photoreceptive. Under such conditions, melanopsin acts as a sensory photopigment, coupled to a native ion channel via a G-protein signalling cascade, to drive physiological light detection. The melanopsin photoresponse relies on the presence of cis-isoforms of retinaldehyde and is selectively sensitive to short-wavelength light. We also present evidence to show that melanopsin functions as a bistable pigment in this system, having an intrinsic photoisomerase regeneration function that is chromatically shifted to longer wavelengths.     

The mPer2 gene encodes a functional component of the mammalian circadian clock.

Circadian rhythms are driven by endogenous biological clocks that regulate many biochemical, physiological and behavioural processes in a wide range of life forms. In mammals, there is a master circadian clock in the suprachiasmatic nucleus of the anterior hypothalamus. Three putative mammalian homologues (mPer1, mPer2 and mPer3) of the Drosophila circadian clock gene period (per) have been identified. The mPer genes share a conserved PAS domain (a dimerization domain found in Per, Arnt and Sim) and show a circadian expression pattern in the suprachiasmatic nucleus. To assess the in vivo function of mPer2, we generated and characterized a deletion mutation in the PAS domain of the mouse mPer2 gene. Here we show that mice homozygous for this mutation display a shorter circadian period followed by a loss of circadian rhythmicity in constant darkness. The mutation also diminishes the oscillating expression of both mPer1 and mPer2 in the suprachiasmatic nucleus, indicating that mPer2 may regulate mPer1 in vivo. These data provide evidence that an mPer gene functions in the circadian clock, and define mPer2 as a component of the mammalian circadian oscillator.     

CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis

Flowering is often triggered by exposing plants to appropriate day lengths. This response requires an endogenous timer called the circadian clock to measure the duration of the day or night. This timer also controls daily rhythms in gene expression and behavioural patterns such as leaf movements. Several Arabidopsis mutations affect both circadian processes and flowering time; but how the effect of these mutations on the circadian clock is related to their influence on flowering remains unknown. Here we show that expression of CONSTANS (CO), a gene that accelerates flowering in response to long days, is modulated by the circadian clock and day length. Expression of a CO target gene, called FLOWERING LOCUS T (FT), is restricted to a similar time of day as expression of CO. Three mutations that affect circadian rhythms and flowering time alter CO and FT expression in ways that are consistent with their effects on flowering. In addition, the late flowering phenotype of such mutants is corrected by overexpressing CO. Thus, CO acts between the circadian clock and the control of flowering, suggesting mechanisms by which day length regulates flowering time.     

谷氨酸、生物节律与运动

生物节律是指生物在自然选择、长期进化过程中保存下来的适应性表象。谷氨酸(Glu)是中枢神经系统(CNS)中主要的兴奋性氨基酸类神经递质,已经证实谷氨酸含量有近似昼夜节律现象,同时谷氨酸在调节人体生物节律的过程中扮演重要角色。最近的研究发现,择时运动对谷氨酸的含量及近似昼夜节律会产生明显影响,对运动队异地训练和比赛时快速地调整时差、延缓疲劳有重要意义。     

Regulation of circadian behaviour and metabolism by synthetic REV-ERB agonists

Synchronizing rhythms of behaviour and metabolic processes is important for cardiovascular health and preventing metabolic diseases. The nuclear receptors REV-ERB-α and REV-ERB-β have an integral role in regulating the expression of core clock proteins driving rhythms in activity and metabolism. Here we describe the identification of potent synthetic REV-ERB agonists with in vivo activity. Administration of synthetic REV-ERB ligands alters circadian behaviour and the circadian pattern of core clock gene expression in the hypothalami of mice. The circadian pattern of expression of an array of metabolic genes in the liver, skeletal muscle and adipose tissue was also altered, resulting in increased energy expenditure. Treatment of diet-induced obese mice with a REV-ERB agonist decreased obesity by reducing fat mass and markedly improving dyslipidaemia and hyperglycaemia. These results indicate that synthetic REV-ERB ligands that pharmacologically target the circadian rhythm may be beneficial in the treatment of sleep disorders as well as metabolic diseases.     

Feedback of the Drosophila period gene product on circadian cycling of its messenger RNA levels

Mutations in the period (per) gene of Drosophila melanogaster affect both circadian and ultradian rhythms. Levels of per gene product undergo circadian oscillation, and it is now shown that there is an underlying oscillation in the level of per RNA. The observations indicate that the cycling of per-encoded protein could result from per RNA cycling, and that there is a feedback loop through which the activity of per-encoded protein causes cycling of its own RNA.