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Manifestation of circadian rhythm under constant light depends on lighting conditions during lactation.

Adult rats transferred to continuous illumination (LL) show a disruption of circadian rhythms, although the mechanisms underlying this effect are not yet well known. In previous experiments, we found that when rats were born and raised under LL they showed an ultradian pattern during the first 10 days after weaning, but afterward they generated a circadian rhythm that was maintained until adulthood. It was not clear whether this evolution was attributable to the influence of the rhythm of the mother or to the effect of constant light. Here, we have studied the motor activity rhythm of young rats maintained under LL after weaning, taking into account the conditions to which they were exposed during lactation [LL or continuous darkness (DD)]. To check the possible effect of the rhythm of the dam, on the day of delivery some of the dams were blinded, others were subjected to a restricted feeding schedule of 3 h/day, and the others were used as controls. For each rat, the period of the circadian rhythm and the percentage of variance explained by this rhythm were calculated. Results show that all rats maintained under LL during lactation expressed a circadian rhythm in their motor activity. However, rats maintained under DD during lactation did not. This effect did not seem to be dependent on the type of dam. These results suggest that the rhythm of the dams does not affect the manifestation of the rhythm of the pups and that the expression of circadian rhythmicity under constant bright light depends on the lighting conditions under which the animals were maintained during lactation, which could affect the development of the circadian pacemaker or the retina.

Activity Cycles↗

Transiently expressed early light-inducible thylakoid proteins share transmembrane domains with light-harvesting chlorophyll binding proteins.

The early light-inducible proteins (ELIPs) of barley chloroplasts are encoded in two multigene families yielding end products of different molecular mass. Sequencing of complete cDNA clones showed that the low and high molecular mass proteins differ by the presence or absence of a 65 amino acid peptide in the amino-terminal part of the mature proteins. Two domains of the ELIPs reveal striking similarity in amino acid sequence with two transmembrane domains of all known light-harvesting chlorophyll a/b-binding proteins from photosystem I and II and may be of importance in anchoring the polypeptides in the membrane. The cDNA sequences of two low molecular mass ELIPs differ by an insert of 5 codons in the putative transit peptide. By in vitro transcription and translation of the cloned DNA and subsequent transport of the products into chloroplasts it could be established that the precursors are processed into products of identical apparent molecular mass. In vitro translated ELIPs were incorporated into thylakoid membranes both as precursors and mature polypeptides. It is suggested that ELIPs are pigment-free substitutes for light-harvesting polypeptides in the assembly of photosynthetic units during early development of thylakoids.

Amino Acid Sequence↗

Luminance detectors in the olivary pretectal nucleus and their relationship to the pupillary light reflex in the rat. II. Studies using sinusoidal light.

The luminance detectors in the olivary pretectal nucleus, which are likely candidates mediating the pupillary light reflex, responded to all frequencies of sinusoidally modulated light up to 12-25 Hz. At low frequencies (0.05-4.0 Hz) the luminance detectors responded with modulated firing to different stimulation rates. The modulation depth of the cell response increased with the increase in stimulation frequency up to 20 Hz, then rapidly fell. There was a delay between the peak intensity of the stimulus and the peak firing of cell response of about 30-40 ms. The amplitudes of the consensual pupil responses to the same sinusoidal stimulus, on the other hand, decreased with an increase in frequency and no discernible response was recorded above 2 Hz. The pupil responses were little affected by sympathectomy. The differences in the frequency response characteristics of luminance detectors and the pupil were attributed to the sluggish dynamic properties of the pupil muscles. This was demonstrated using an electronic model of the iris muscle which modified the responses of the luminance detectors giving output waveforms broadly resembling pupil responses to square and sinusoidally modulated lights.

Animals↗

The influence of P-light chain phosphorylation by myosin light chain kinase on the calcium sensitivity of chemically skinned heart fibres.

Phosphorylation of the P-light chain of myosin might be involved in the regulation of cardiac contractility. Thus an enhanced phosphorylation level of the P-light chain catalyzed by Ca2+-calmodulin-dependent myosin light chain kinase (MLCK) increased significantly the Ca2+ sensitivity of chemically skinned ventricular fibre bundles of the pig. This effect was reversible. Whereas force development at submaximal Ca2+ concentration (pCa 5.5) increased by approximately 50% in the presence of MLCK, maximum tension achieved at maximum Ca2+-concentration (pCa 4.3) was not affected.

Animals↗

Ultrasound-induced light transmission change within platelet suspensions as an indirect measure of platelet morphology: preliminary correlation with stirring-induced light transmission change.

Discoid platelets were directionally orientated using stirring or ultrasound, and the change from random orientation was measured by light transmission. Stirring-induced light transmission change (SILTC), already well established as a good indirect indicator of platelet morphology, was compared with ultrasound-induced light transmission change (USILTC) in order to determine whether USILTC may be a similarly good indicator of platelet morphology. Numerous direct comparisons were made between SILTC and USILTC at various static temperatures, and during the dynamic thermal transitions of warming and cooling. Good correlation exists between quantitated SILTC and USILTC values, but the relationship is not exact. Furthermore, single stirring events may considerably distort the relationship under certain conditions. At the ultrasound frequency of 5 MHz and at an estimated peak positive pressure of 2.03 X 10(4) Pa, the USILTC response patterns exhibit a qualitative "spike" pattern for unfixed platelets, which disappears following glutaraldehyde fixation. These preliminary findings are described and discussed in detail, since they suggest that USILTC is a better index of platelet viability than SILTC, although much research is still required in this area.

Biophysical Phenomena↗

Analytical performance of serum free light-chain assay during monitoring of patients with monoclonal light-chain diseases.

BACKGROUND: Measurement of serum free light chains (FLC) is useful for the diagnosis and monitoring of monoclonal light-chain diseases. It has been suggested that there will be widespread replacement of urine Bence Jones protein measurement by serum FLC assay. We report on our experience with the assay during monitoring of light-chain myeloma (LCMM) and AL amyloidosis (AL). METHODS: Serum FLC immunoassay, serum and/or urine protein electrophoresis and immunofixation were performed on serial samples during monitoring of LCMM. Recovery and immunoreactivity of FLC were tested by sample dilution. Assay precision was determined by repeat assay of samples over several reagent lots. RESULTS: In one of 23 patients with LCMM there was non-reaction of a monoclonal kappa FLC with some reagent lots and the assay did not indicate disease relapse. Samples showed non-linear, non-parallel immunoreactivity on dilution. Several tested monoclonal FLC gave lower values at the assay starting dilution compared with higher sample dilution and non-parallel dose-response curves. The median between-reagent lot variation for FLC measurement was 19-20% CV. CONCLUSIONS: Laboratory staff and clinicians need to be aware of the potential for non-reactivity of individual monoclonal FLC, and the effects of dilution and precision on FLC values and their interpretation.

Aged↗

Ion transport across model lipid membranes containing light-harvesting complex II: an effect of light.

The effect of light on proton transport across lipid membranes of small unilamellar liposomes containing incorporated major light-harvesting pigment-protein complex of Photosystem II (LHCII) has been studied with the application of pH-sensitive dyes entrapped inside vesicles. Proton permeability coefficient for LHCII-modified membranes was found to be about twice as high as in the case of the control pure lipid vesicles. Illumination of the samples with light absorbed by the LHCII-bound photosynthetic pigments considerably affects the kinetics of proton transport: it increases the rate and decreases the steady-state level of proton gradient across the membranes. The effect was interpreted in terms of heat-induced conformational changes of LHCII molecular structures that affect proton buffering capacity of this protein. Both the control and the LHCII-modified lipid membranes have been found to be practically impermeable to Ca(++) ions, as demonstrated by fluorescence of liposome-entrapped calcium-sensitive probe calcium crimson. The slight differences in the proton transport across the LHCII-containing membranes under the presence of Ca(++) suggest calcium binding to this antenna protein.

Animals↗

Shedding light on plant competition: modelling the influence of plant morphology on light capture (and vice versa).

A plant's morphology is both strongly influenced by local light availability and, simultaneously, strongly influences this local light availability. This reciprocal relationship is complex, but lies at the heart of understanding plant growth and competition. Here, we develop a sub-individual-based simulation model, cast at the level of interacting plant components. The model explicitly simulates growth, development and competition for light at the level of leaves, branches, etc., located in 3D space. In this way, we are able to explore the manner in which the low-level processes governing plant growth and development give rise to individual-, cohort-, and community-level phenomena. In particular, we show that individual-level trade-offs between growing up and growing out arise naturally in the model, and robustly give rise to cohort-level phenomena such as self-thinning, and community processes such as the effect of ecological disturbance on the maintenance of biodiversity. We conclude with a note on our methodology and how to interpret the results of simulation models such as this one.

Algorithms↗

Blue light regeneration of bacteriorhodopsin bleached by continuous light.

Photobleaching of bacteriorhodopsin (BR) by continuous light has recently been demonstrated. This bleaching consists of at least two subsequent product states. One of them is absorbing maximally in the blue spectral region. Our present study shows that upon illumination of the bleached sample with blue light a back photoprocess appears, resulting in regeneration of the original BR state. From a technical point of view, the observed phenomenon is similar to the reverting effect of blue light on the photocycle. An important difference is that the photobleached state of BR is much more stable than any of the photocycle intermediates, and may provide an advantage for several technical applications.

Bacteriorhodopsins↗

Light perception in plants: cytokinins and red light join forces to keep phytochrome B active.

Plant growth and development is modulated by internal cues such as rhe hormonal balance and external factors. Plants are particularly sensitive to their light environment, which they scrutinize with at least three classes of photoreceptors. In recent years, it has become increasingly clear that light and hormonal signaling interact at several levels. A cytokinin receptor was recently identified together with several elements acting in this signaling pathway. ARR4, a response regulator working downstream of a cytokinin receptor, has been shown to regulate phytochrome B-mediated light signaling.

Arabidopsis Proteins↗

Light deprivation suppresses the light response of inner retina in both young and adult mouse.

The retinal synaptic network continues its development after birth in mammals. Recent studies show that postnatal development of retinal circuitry depends on visual stimulation. We sought to determine whether there is a time period during which the retina shows evidence of increased plasticity. We examined the effects of light deprivation on the retinal light response of mouse retina using electroretinogram (ERG) measurements. Our results showed that dark rearing mice from birth to postnatal day (P) 30, 60, and 90 suppressed the amplitudes of oscillatory potentials (OPs) and the magnitudes of suppression were age independent. In addition, dark-rearing-produced suppression of OP amplitudes can be completely reversed in both young and adult mice by returning them to cyclic light/dark conditions for 1 to 2 weeks. However, the recovery time course was age dependent with younger animals needing a longer time to achieve a full recovery. Furthermore, dark rearing of P60 mice raised under cyclic light/dark conditions for 30 days resulted in a similar magnitude of suppression of OP amplitudes as in age-matched mice dark reared from birth. These findings demonstrate that both the normal developmental changes and the maintenance of mature inner retinal light response in adult animals require visual stimulation. These results indicate a degree of activity-dependent plasticity in mouse retina that has not been previously described.

Aging↗

Light-regulated biochemical events in invertebrate photoreceptors. 1. Light-activated guanosinetriphosphatase, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes.

The occurrence of a guanine nucleotide binding protein activated by squid rhodopsin was established by examination of GTPase activity, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes. Purified squid (Loligo opalescens) photoreceptors exhibited GTPase activity that increased 3-4-fold by illumination. Half-maximal GTPase activity was observed when 2% of the rhodopsin was photoconverted to metarhodopsin. The Km of the light-regulated activity was 1 microM GTP. Binding of the hydrolysis-resistant GTP analogue guanosine 5'-(beta, gamma-imidotriphosphate) [Gpp(NH)p] was enhanced greater than 10 times by illumination. A protein, Mr 44 000, was identified as a component of the light-activated guanine nucleotide binding protein/GTPase through its specific labeling with [32P]NAD catalyzed by cholera toxin: light increased the extent of 32P incorporation 7-fold. The addition of ATP to the membrane suspension enhanced labeling, while guanine nucleotides inhibited labeling with the relative potency GTP gamma S much greater than GDP greater than GTP greater than Gpp(NH)p. The 44 000-dalton protein was membrane bound irrespective of variations in ionic strength and divalent ion concentration over a wide range. These results suggest that a G protein, which incorporates both GTP binding and hydrolysis functions, is intimately involved in the visual process of invertebrate photoreceptors.

Animals↗

Light-induced glutamate transport in Halobacterium halobium envelope vesicles. I. Kinetics of the light-dependent and the sodium-gradient-dependent uptake.

During illumination Halobacterium halobium cell envelope vesicles accumulate [3H]glutamate by an apparently unidirectional transport system. The driving force for the active transport originates from the light-dependent translocation of protons by bacteriorhodopsin and is due to a transmembrane electrical potential rather than a pH difference. Transport of glutamate against high concentration gradients is also achieved in the dark, with high outside/inside Na+ gradients. Transport in both cases proceeds with similar kinetics and shows a requirement for Na+ on the outside and for K+ on the inside of the vesicles. The unidirectional nature of glutamate transport seems to be due to the low permeability of the membranes to the anionic glutamate, and to the differential cation requirement of the carrier on the two sides of the membrane for substrate translocation. Thus, glutamate gradients can be collapsed in the dark either by lowering the intravesicle pH (with nigericin, or carbonyl cyanide p-trifluoromethoxyphenylhydrazone plus valinomycin), or by reversing the cation balance across the membranes, i.e., providing NaCl on the inside and KCl on the outside of the vesicles. In contrast to the case of light-dependent glutamate transport, the initial rates of Na+-gradient-dependent transport are not depressed when an opposing diffusion potential is introduced by adding the membrane-permeant cation, triphenylmethylphosphonium bromide. Therefore, it appears that, although the electrical potential must be the primary source of energy for the light-dependent transport, the translocation step itself is electrically neutral.

Biological Transport, Active↗

Vibration spectroscopy reveals light-induced chromophore and protein structural changes in the LOV2 domain of the plant blue-light receptor phototropin 1.

Phototropins (phot1 and phot2), the plant blue-light receptors for phototropism, chloroplast movement, and stomatal opening, are flavoproteins that contain two approximately 12 kDa FMN-binding domains, LOV1 and LOV2, at their N-terminus, and a serine/threonine protein kinase domain at their C-terminus. The light-activated LOV2 domain forms a metastable intermediate which has been shown to be a protein-chromophore cysteinyl adduct (Cys39) at C(4a) of FMN. This species thermally relaxes back to the ground state in the dark. We measured the light-minus-dark FTIR difference spectra for the LOV2 domain of oat phot1. These spectra show the disappearance of bands at 1580, 1550, and 1350 cm(-1) that originate from, or are strongly coupled to, the N5=C(4a) stretching vibrations, consistent with the perturbations expected upon C(4a) adduct formation. Assignment of these negative difference FTIR bands to native chromophore vibrations is based on the alignment with resonance Raman bands of FMN. Prominent positive bands include a doublet at 1516 and 1536 cm(-1) and one at 1375 and 1298 cm(-1). Normal-mode vibrational-frequency calculations for both lumiflavin and lumiflavin with a sulfur attached at the C(4a) position agree with many of the positive and negative bands observed in the difference spectra. Both calculated and experimental difference FTIR spectra for deuterium isotope substitutions at exchangeable positions in the flavin chromophore are consistent with the assignment of the above positive bands to vibrational modes involving both the newly formed tetrahedral geometry of C(4a) and the N5-H bond in the long-lived LOV2(S)(390) cysteinyl species.

Amides↗

Xanthophylls in light-harvesting complex II of higher plants: light harvesting and triplet quenching.

A spectral and functional assignment of the xanthophylls in monomeric and trimeric light-harvesting complex II of green plants has been obtained using HPLC analysis of the pigment composition, laser-flash induced triplet-minus-singlet, fluorescence excitation, and absorption spectra. It is shown that violaxanthin is not present in monomeric preparations, that it has most likely a red-most absorption maximum at 510 nm in the trimeric complex, and that it is involved in both light-harvesting and Chl-triplet quenching. Two xanthophylls (per monomer) have an absorption maximum at 494 nm. These play a major role in both singlet and triplet transfer. These two are most probably the two xanthophylls resolved in the crystal structure, tentatively assigned to lutein, that are close to several chlorophyll molecules [Kühlbrandt, W., Wang, N. D., & Fujiyoshi, Y. (1994) Nature 367, 614-621]. A last xanthophyll contribution, with an absorption maximum at 486 nm, does not seem to play a significant role in light-harvesting or in Chl-triplet quenching. On the basis of the assumption that the two structurally resolved xanthophylls are lutein, this 486 nm absorbing xanthophyll should be neoxanthin. The measurements demonstrate that violaxanthin is connected to at least one chlorophyll a with an absorption maximum near 670 nm, whereas the xanthophylls absorbing at 494 nm are connected to at least one chlorophyll a with a peak near 675 nm.

Chromatography, High Pressure Liquid↗

AL-amyloidosis and light-chain deposition disease light chains induce divergent phenotypic transformations of human mesangial cells.

Human mesangial cells (HMCs) are injured by either excessive amounts or abnormal light chains (LCs), or a combination of both in patients with plasma cell dyscrasias. Consequently, these HMCs undergo phenotypic transformations. HMCs were incubated with eight different light-chains (LCs) for 96 h. These cells, in addition to 51 patient samples from patients with AL-amyloidosis (AL-Am), light-chain deposition disease (LCDD), myeloma cast nephropathy (MCN) and controls were analyzed by immunohistochemistry for CD68, muscle-specific actin (MSA), smooth muscle actin (SMA), CD14, and Ham56 protein expressions. All samples were also studied using electron microscopy. Greater staining (four- and three-fold) expressions of CD68 and Ham56, respectively, were observed in the HMCs incubated with AL-Am-LCs compared to those with LCDD-LCs and control. SMA expression levels were five-fold higher in LCDD-LC-treated cells compared to the other categories of LC-treated and control cells. Similar results were obtained in the renal specimens, however, CD68 levels were 12-fold higher in the AL-Am cases compared to the LCDD cases, respectively. Conversely, MSA and SMA levels were three fold higher in the LCDD cases than in the AL-Am ones. No CD14 expression was noted in any of the samples and CD-34 staining of HMCs treated with the various LCs only showed rare positive cells. Dynamic real-time studies to visualize the rough endoplasmic reticulum (RER) and lysosomal compartments in HMCs incubated with LCDD and AL-Am-LCs showed striking expansion of each of the above-mentioned compartments, respectively. This indicates the presence of more RER in the LCDD-LC-treated HMCs and a striking increase in lysosomes noticeable in the AL-Am-LC-treated cells. Data obtained in this study highlighted that HMCs incubated with LCDD-LCs undergo a myofibroblastic phenotypic transformation, while AL-Am-LCs induce a macrophage-like phenotype in these cells.

Actins↗

Mutations throughout an Arabidopsis blue-light photoreceptor impair blue-light-responsive anthocyanin accumulation and inhibition of hypocotyl elongation.

This paper reports the characterization of novel mutations within the Arabidopsis thaliana HY4 gene, which has previously been shown to encode a protein (CRY1) with characteristics of a blue-light photoreceptor. Several point mutations were identified within the amino-terminal domain of CRY1--this region of CRY1 has high homology to photolyase and is likely to be involved in blue-light-mediated electron transfer. Mutations were found within the region of homology to the known chromophore binding domains of photolyase. Point mutations within the 200 amino acid carboxy-terminal extension distinguishing CRY1 from photolyase, likewise disrupt function of the protein. CRY1 was originally defined as the photoreceptor responsible for blue-light-mediated inhibition of hypocotyl elongation and we now report that anthocyanin accumulation in germinating seedlings is an additional phenotype under the control of this photoreceptor--this is shown to be mediated in part by modulation of mRNA levels of chalcone synthase, one of the anthocyanin biosynthetic enzymes. The effect of the novel mutations on both inhibition of hypocotyl elongation and anthocyanin biosynthesis have been evaluated, and it is demonstrated that mutations with less severe effects on hypocotyl elongation show a similarly reduced effect on anthocyanin biosynthesis. These results are consistent with the cryptochrome photoreceptor mediating multiple regulatory pathways by the same primary mode of action.

Amino Acid Sequence↗

Structural analysis of photosystem II in far-red-light-adapted thylakoid membranes. New crystal forms provide evidence for a dynamic reorganization of light-harvesting antennae subunits.

We studied two-dimensional crystals of the major pigment-protein complex, photosystem II, in far-red-light-adapted thylakoid membranes of the viridis-zb63 mutant of barley. Significantly larger grana membranes were produced with an increased synthesis of the entire photosystem II complex. These red-light-adapted membranes also contained two-dimensional crystals with a high frequency. Three different crystal forms of photosystem II were observed, providing the following data which further our understanding of the architecture of the native complex. (a) The oligomeric form of photosystem II in the membrane was monomeric in all crystal forms, but with a clear non-crystallographic pseudo-twofold symmetry. This was more apparent on the lumenal face of the complex. (b) The variability of unit cell contacts in different crystal forms implied that the peripheral light-harvesting antenna complex and the core of the complex were loosely connected. These peripheral subunits were predicted to rearrange so that they can either encircle the core complex or associate in parallel channels separated by lines of core complexes. (c) Grana membranes were found to retain a double-layered inside-out character, with a stromal face-to-stromal face packing. However, the presence of a crystal in one membrane did not necessarily impose crystallinity on its pair.

Adaptation, Physiological↗