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DARPP-32 involvement in the photic pathway of the circadian system.

The multifunctional regulator of protein kinases and phosphatases dopamine- and cAMP-regulated phosphoprotein of 32 kDa (DARPP-32) is an important molecular target of the dopamine signaling pathway. In the present study, we investigated the possible involvement of DARPP-32 regulation in the circadian system using DARPP-32 knock-out (KO) mice. These mice showed normal entrainment to a 12 h light/dark cycle and free run in constant darkness with a period similar to that of wild-type controls. After light exposure, however, the behavioral phase-delay response and the expression of light-induced clock gene mPer2 were attenuated in the DARPP-32 KO mice. Attenuated phase delays were also seen in animals bearing a point mutation in DARPP-32 at the PKA (Thr34) but not at the casein kinase I (Ser130) phosphorylation site. We next examined DARPP-32 expression in the retina and intergeniculate leaflet (IGL), both of which convey light information to the suprachiasmatic nucleus (SCN), the locus of a master circadian clock, and in the SCN itself. DARPP-32 was expressed in the retina but not in the IGL or the SCN. The results indicate that DARPP-32 is involved in the retinal pathway transmitting photic information that resets the circadian clock.

Animals↗

Spectral organization of ommatidia in flower-visiting insects.

This article reviews recent advances of studies on the spectral organization of the compound eye in lepidopteran and hymenopteran insects. The compound eyes consist of ommatidia, which contain a set of photoreceptor cells. The common feature is that the ommatidia can be divided into three types, according to the combination of spectral classes of photoreceptors. Honeybees and nymphalid butterflies provide the simplest case with three photoreceptor classes having peak sensitivities in the ultraviolet (UV), blue (B), or green (G) wavelength region. These receptors populate the ommatidia in fixed combinations. In type I ommatidia, the main eight photoreceptors consist of one UV, one B, and six G receptors. Type II has two UV and six G receptors, and type III has two B and six G receptors. The organization is basically retained in all insect species studied so far, although some butterflies have more than six classes of spectral receptors, including those sensitive in the violet and red wavelength regions. To acquire these additional receptors, two distinct strategies are applied: the multiple opsin strategy, taken by the Japanese yellow swallowtail butterfly Papilio xuthus, and the filter strategy, used by the small white butterfly Pieris rapae.

Animals↗

High light inhibits chlorophyll biosynthesis at the level of 5-aminolevulinate synthesis during de-etiolation in cucumber (Cucumis sativus) cotyledons.

Using the vascular plant Cucumis sativus (cucumber) as a model, we studied the effects of high (intense and excess) light upon chlorophyll biosynthesis during de-etiolation. When illuminated with high light (1500-1600 microE/m2/s), etiolated cucumber cotyledons failed to synthesize chlorophyll entirely. However, upon transfer to low light conditions (40-45 microE/m2/s), chlorophyll biosynthesis and subsequent accumulation resumed following an initial 2-12 h delay. Duration of high light treatment negatively correlated with chlorophyll biosynthetic activity. Specifically, we found that high light severely inhibited 5-aminolevulinic acid (ALA) synthesis. This effect partly could be because of the decrease in protein level of glutamyl-tRNA reductase (GluTR) observed. Protein level of glutamate-1-semialdehyde (GSA-AT) remained unchanged. It was also found that high light did not suppress HEMA 1 expression. Therefore, we speculated that this significant inhibition of ALA synthesis might have occurred mainly because of concomitant inactivation of GluTR and/or inhibition of complex formation between GluTR and GSA-AT. Our further observation that both methyl viologen and rose bengal similarly inhibit ALA synthesis under low light conditions suggested that reactive oxygen species (ROS) could be responsible for the inhibition of ALA synthesis in cotyledons exposed to high light conditions.

Aldehyde Oxidoreductases↗

Additivity in murine circadian phototransduction.

Additivity in the circadian phototransduction system of the mouse has not been tested directly. Because of this, accurate prediction of circadian phase shifts elicited by polychromatic light stimuli cannot be derived from the results of studies using monochromatic light stimuli. This limitation also makes it impossible to deduce the relative contributions of the photoreceptive mechanisms (rods, cones and melanopsin-containing retinal ganglion cells) underlying circadian phototransduction in the mouse. Using nearly monochromatic light stimuli of different spectral composition, and combinations thereof, we demonstrated that murine circadian phototransduction exhibits additivity. Based on the locomotor activity phase shifts elicited by these stimuli, we developed the first quantitative assessment of the relative contributions of conventional and novel photoreceptive mechanisms for circadian functioning in the mouse.

Animals↗

Bilirubin, REM sleep, and phototransduction of environmental time cues. A hypothesis.

The prevailing hypothesis for phototransduction is that visual (rod or cone) pigments mediate light's primary effects on biological clock systems. Common light-responsive chronobiological behavioral properties of plants and animals and some common molecular structures of plants and animals suggest the possibility that heme moieties and bile pigments in animals mediate some nonvisual influences of light on neuroactive gases and biological rhythms. As plant phytochrome resets the plant biological clock, the similar chromophore in bile pigments is proposed to transduce environmental light zeitgeber signals to endogenous biological clocks. The temporal association of plasma bilirubin and rapid-eye-movement (REM) sleep in populations, the correlation of secretion of biliary bilirubin with REM sleep among 10 different species (Spearman r = 0.89, p < 0.002), and the known responses of bilirubin to light lead to the hypothesis that bilirubin, in particular, plays an evolutionary role in the regulation of REM sleep and in mediating some of light's antidepressant effects.

Animals↗

The insulin receptor: structure and function.

Promising progress in understanding the molecular basis of insulin action has been achieved by demonstrating that the insulin receptor is an insulin-sensitive tyrosine kinase. Here we discuss the structure of this receptor kinase and compare it with receptors for related growth factors. We review the known modes to regulate the receptor kinase activity, either through its autophosphorylation (on tyrosine residues) or through its phosphorylation by other kinases (on serine and threonine residues). We discuss the role of the receptor kinase activity in hormone signal transduction in light of results indicating a reduced kinase activity in insulin-resistant states. Finally, studies to identify natural substrates for the insulin receptor kinase are presented. The possible physiological role of these phosphorylated substrates in mediating insulin action is evaluated.

Animals↗

New techniques in fast time-resolved structure determination.

New techniques in fast time-resolved X-ray crystallography provide a different approach to understanding the structural basis of protein function. Two biological systems have been studied as part of the refinement of these techniques, and have actually spurred new ideas in time-resolved structural studies. The dissociation of carbon monoxide from carbon-monoxy myoglobin has earlier been investigated over a time range spanning 18 orders of magnitude (femtoseconds to hours) using spectroscopic methods. Rapid time-resolved determination of the entire myoglobin structure made it possible to determine both the position of the CO after photodissociation and the entire globin structure, over a time range from nanoseconds to milliseconds, during which the heme and globin relax and the carbon monoxide rebinds. Photoactive yellow protein, a relative newcomer to biophysical research, has a fully-reversible photocycle containing several spectrally distinct intermediates. Identifying and solving the structures of each intermediate is the initial goal in time-resolved studies on this protein and will contribute to a greater understanding of the biological process of light driven signal transduction.

Animals↗

Opsin-G11-mediated signaling pathway for photic entrainment of the chicken pineal circadian clock.

Light is a major environmental signal for entrainment of the circadian clock, but little is known about the intracellular phototransduction pathway triggered by light activation of the photoreceptive molecule(s) responsible for the phase shift of the clock in vertebrates. The chicken pineal gland and retina contain the autonomous circadian oscillators together with the photic entrainment pathway, and hence they represent useful experimental models for the clock system. Here we show the expression of G11alpha, an alpha subunit of heterotrimeric G-protein, in both tissues by cDNA cloning, Northern blot, and Western blot analyses. G11alpha immunoreactivity was colocalized with pinopsin in the chicken pineal cells and also with rhodopsin in the outer segments of retinal photoreceptor cells, suggesting functional coupling of G11alpha with opsins in the clock-containing photosensitive tissues. The physical interaction was examined by coimmunoprecipitation experiments, the results of which provided evidence for light- and GTP-dependent coupling between rhodopsin and G11alpha. To examine whether activation of endogenous G11 leads to a phase shift of the oscillator, Gq/11-coupled m1-type muscarinic acetylcholine receptor (mAChR) was ectopically expressed in the cultured pineal cells. Subsequent treatment of the cells with carbamylcholine (CCh), an agonist of mAChR, induced phase-dependent phase shifts of the melatonin rhythm in a manner very similar to the effect of light. In contrast, CCh treatment induced no measurable effect on the rhythm of nontransfected (control) cells or cells expressing G(i/o)-coupled m2-type mAChR, indicating selectivity of the G-protein activation. Together, our results demonstrate the existence of a G11-mediated opsin-signaling pathway contributing to the photic entrainment of the circadian clock.

Animals↗

Mouse cone arrestin expression pattern: light induced translocation in cone photoreceptors.

PURPOSE: Arrestins are a superfamily of regulatory proteins that down-regulate activated and phosphorylated G-protein coupled receptors (GPCRs). Cone arrestin (CAR) is expressed in cone photoreceptors and pinealocytes and may contribute to the shutoff mechanisms associtated with high acuity color vision. To initiate a study of CAR's function in cone phototransduction, the mouse CAR (mCAR) transcript and protein expression patterns are examined and in vitro binding assays are also presented. METHODS: Tissue distribution of mCAR was determined by Northern and immunoblot analyses and its cellular localization identified by In situ hybridization and immunohistochemistry. The protein expression pattern of mCAR in the postnatal developmental and adult mouse retina was analyzed by immunoblotting in normal C57 and rd/rd mouse retinas. In vitro binding assays with in vitro translated arrestins were used to study the interaction of mCAR and mouse S-antigen (mSAG) with embryonic chicken outer segment (OS) membranes containing both rod and cone opsins. RESULTS: MCAR has a high level of amino acid sequence identity with orthologous sequences reported for other species except the C-terminal region, which is highly conserved between mouse and rat but divergent in other species. MCAR is expressed exclusively in the retina and the pineal gland, and unique isoforms are expressed during postnatal development of the retina and the pineal gland. The postnatal developmental expression pattern of mCAR and mSAG in the rd/rd mouse retina parallels the generation and degeneration of the cone and rod photoreceptors in these mice. In situ and immunohistochemistry both reveal cone-specific expression of mCAR in the retina. Immunofluorescent staining of retinal sections from dark-adapted or light-exposed mice suggests a light-dependent translocation of mCAR immunoreactivity from the cone inner segments (CIS) and other parts of the cell body to the cone outer segments (COS), similar to but not as dramatic as rod arrestin. In vitro binding assays show a small yet significant increase in binding of the full-length mCAR (mCARFL) to embryonic chicken OS membranes following light activation and phosphorylation of the opsins in the membranes. CONCLUSIONS: MCAR is expressed in retinal cone photoreceptors and the pineal gland. The light-dependent translocation of mCAR immunoreactivity and the increase of mCAR binding to light-activated, phosphorylated embryonic chicken OS membranes, compared to its binding to dark, unphosphorylated membranes, suggest the possibility that mCAR is involved in shutting off the phototransduction cascade in cone photoreceptors as rod arrestin does in rod photoreceptors. However, prominent differences exist between rod arrestin and CAR, suggesting other functions for CAR.

Animals↗

A broad role for melanopsin in nonvisual photoreception.

The rod and cone photoreceptors that mediate visual phototransduction in mammals are not required for light-induced circadian entrainment, negative masking of locomotor activity, suppression of pineal melatonin, or the pupillary light reflex. The photopigment melanopsin has recently been identified in intrinsically photosensitive retinal ganglion cells (RGCs) that project to the suprachiasmatic nucleus (SCN), intergeniculate leaflet (IGL), and olivary pretectal nucleus, suggesting that melanopsin might influence a variety of irradiance-driven responses. We have found novel projections from RGCs that express melanopsin mRNA to the ventral subparaventricular zone (vSPZ), a region involved in circadian regulation and negative masking, and the sleep-active ventrolateral preoptic nucleus (VLPO) and determined the subsets of melanopsin-expressing RGCs that project to the SCN, the pretectal area (PTA), and the IGL division of the lateral geniculate nucleus (LGN). Melanopsin was expressed in the majority of RGCs that project to the SCN, vSPZ, and VLPO and in a subpopulation of RGCs that innervate the PTA and the IGL but not in RGCs projecting to the dorsal LGN or superior colliculus. Two-thirds of RGCs containing melanopsin transcript projected to each of the SCN and contralateral PTA, and one-fifth projected to the ipsilateral IGL. Double-retrograde tracing from the SCN and PTA demonstrated a subpopulation of RGCs projecting to both sites, most of which contained melanopsin mRNA. Our results suggest that melanopsin expression defines a subset of RGCs that play a broad role in the regulation of nonvisual photoreception, providing collateralized projections that contribute to circadian entrainment, negative masking, the regulation of sleep-wake states, and the pupillary light reflex.

Animals↗

The possible role of electromagnetic fields in bacterial communication.

This paper presents a general review of the current knowledge regarding bacterial communication via electromagnetic fields. The possible role of ultraviolet, visible and infrared light, extremely high frequency and low frequency electromagnetic fields as well as sound waves is discussed. The probable mechanisms of remote microbial interactions are raised. Future trends in the area of microbiology are also viewed.

Acoustics↗

Role of the pineal organ in the photoregulated hatching of the Atlantic halibut.

The timing of hatching in the Atlantic halibut (Hippoglossus hippoglossus) has been suggested to be regulated by environmental light conditions. However, the photosensory organ that perceives the triggering light has not been identified. In the present study, we investigated the morphogenesis of the pineal organ and the neurochemical differentiation of photoreceptors in the pineal organ and the retina of the Atlantic halibut during embryonic development. Immunocytochemical techniques were used for detection of integral protein components of the phototransduction process: opsins, arrestin (S-antigen) and alpha-transducin. We also studied the expression of serotonin (5-HT), a precursor of the neurohormone melatonin known to be synthesized by pineal photoreceptors. In the pineal anlage, opsin immunoreactive (ir) cells appear at 11 days post-fertilization (pf), arrestin, alpha-transducin and serotonin ir cells appear at 14 days pf; hatching took place 15 days pf. The retina contained no immunoreactive cells in embryos or in newly hatched larva. During this period, the pineal anlage is morphologically discernible only as a wedge-shaped region in the diencephalic roof, where elongated cells are aligned with their long axes converging toward a centrally located presumptive pineal lumen. The results show that the pineal photoreceptors contain serotonin and molecules involved in the phototransduction cascade before hatching. We suggest that the pineal organ has the capacity to perceive and mediate photic information before hatching in halibut embryos, and may thereby influence the timing of hatching.

Animals↗

Photic regulation of pineal function. Analogies between retinal and pineal photoreception.

Light absorbed by a photopigment in a photoreceptor cell causes a photochemical reaction converting the 11-cis retinal chromophore into the all-trans configuration. These changes lead to a series of events that causes cGMP hydrolysis, a following decrease of cGMP in the cytoplasm of the photoreceptor outer segment and a closure of cGMP-gated cationic channels. As a consequence of these processes the membrane hyperpolarizes. In pineal photoreceptor cells of lower vertebrates these processes are only partly investigated. Molecules involved in the phototransduction process and the desensitization, like opsin, vitamin A, alpha-transducin and arrestin, have been immunocytochemically localized in pineal photoreceptors and also electrophysiological studies have shown that phototransduction mechanisms in pineal photoreceptors might be very similar to those found in retinal photoreceptors. This review will summarize some of the current knowledge on pineal photoreception and compare it with retinal processes.

Animals↗

Requirement for the NINAC kinase/myosin for stable termination of the visual cascade.

Activation of the Drosophila photoresponse is a rapid process that results in plasma membrane Ca2+ and Na+ conductances. Ca2+ functions in negative feedback regulation of Drosophila vision including deactivation. Protein kinase C (PKC) binds directly to Ca2+ and is required for deactivation. However, the consequences of disrupting phosphorylation of any individual PKC substrate in the Drosophila retina have not been addressed. In the current work, we show that NINAC p174, which consists of a protein kinase domain joined to the head region of myosin heavy chain, is a phosphoprotein and is phosphorylated in vitro by PKC. Mutation of either of two PKC sites in the p174 tail resulted in an unusual defect in deactivation that had not been detected previously for other ninaC alleles or other loci. After cessation of the light stimulus, there appeared to be a transient reactivation of the visual cascade. This phenotype suggests that a mechanism exists to prevent reactivation of the visual cascade and that p174 participates in this process.

Animals↗