Pigment Migration Research Articles

Pigment‐dispersing hormone‐like peptide in the nervous system of the flies Phormia and Drosophila: Immunocytochemistry and partial characterization

beta-pigment-dispersing hormone (beta-PDH) isolated from the fiddler crab (Rao et al., '85) is a member of an octadecapeptide family of neuropeptides common to arthropods. Whereas earlier studies of these peptides in insects were limited to orthopterans, this investigation focuses on dipteran flies. Extracts of heads from the blowfly Phormia terraenovae were assessed in a fiddler crab bioassay for PDH activity. Immunocytochemistry, dose-response curves, gel filtration chromatography and reversed-phase HPLC, combined with bioassay and enzyme-linked immunosorbent assay (ELISA), indicate the presence of PDH-like peptide in the blowfly. Immunocytochemical mapping of PDH-like immunoreactive (PDHLI) neurons was performed for the entire nervous systems of Phormia and the fruitfly Drosophila with a beta-PDH antiserum. In the cephalic ganglion (brain, optic lobe and subesophageal ganglion) PDHLI cell bodies could be detected (34 in Phormia and 16 in Drosophila). In both species, each hemisphere contains 8 PDHLI cell bodies in the optic lobes. These innervate the optic lobe neuropils bilaterally. In Phormia, another set of 8 cell bodies are located in each of the lateral neurosecretory cell groups in the superior protocerebrum. These neurons send axons to the corpora cardiaca-hypocerebral ganglion complex and to portions of the foregut. In contrast, only the optic lobe neurons display immunoreactivity in Drosophila. Except for the optic lobes, PDHLI processes are distributed only in nonglomerular neurophils of the brain of both species. In the fused thoracico-abdominal ganglia of Phormia, 28 PDHLI cell bodies were found (only six were found in Drosophila). In both species, six abdominal PDHLI neurons are efferents with axons innervating the hindgut. We also found that some of the PDHLI neurons in the Phormia brain and abdominal ganglion contain colocalized FMRFamide-like immunoreactivity. Since the flies studied here do not display hormonally controlled, fast pigment migrations, the PDH-like peptide may have a role as neurotransmitter or neuromodulator in the central nervous system, especially in the visual system, and a regulatory role in the stomatogastric system and the hind-gut.

Neuromodulation of pigment movement in the RPE of normal and 6-OHDA-lesioned goldfish retinas.

The role of dopamine as the endogenous signal-initiating light-dependent changes in the distribution of pigment granules in goldfish retinal pigment epithelium was investigated. In normal retinas, light adaptation resulted in the dispersion of pigment granules. This effect of light was mimicked by the intraocular injection of dopamine or serotonin, which is thought to increase endogenous dopamine release, into dark-adapted eyes. The effect of light, dopamine, or serotonin on dark-adapted retinas was blocked by the dopamine receptor antagonists haloperidol and sulpiride. However, lesioning the endogenous source of retinal dopamine, by prior intraocular injection of 6-hydroxydopamine (6-OHDA), did not block the dispersion of pigment granules in light-adapted retinas. No significant differences in pigment dispersion were noted between unlesioned and lesioned light- or dark-adapted retinas. However, the effect of light on pigment dispersion was no longer blocked by haloperidol or sulpiride in 6-OHDA lesioned animals. Dopamine and serotonin mimicked the effect of light when injected into lesioned dark-adapted eyes, but their effects were also not blocked by haloperidol or sulpiride. These results suggest that dopamine, acting on D2 receptors, is sufficient to induce pigment migration in unlesioned animals. In 6-OHDA-lesioned animals, however, pigment migration is mediated by a receptor mechanism other than D2.

Microwave-Induced Retinal Destruction With Sparing of Sclera and Choriocapillaris

ABSTRACT We studied the effect of short-term hyperthermia on sclera, choroid, and retina by delivering microwave radiation (2.45 GHz) for 1 minute to 12 eyes of Dutch belted pigmented rabbits. Four eyes each were treated with 430C, 450C, and 470C and followed for 4 weeks. The 430C group showed minimal disruption of retinal pigment epithelium and outer retina, with pigment migration; the 450C and 470C groups showed complete retinal and RPE damage, pigment migration, and glial proliferation. At the same time, the sclera and choroid in all of the eyes remained essentially unchanged. We conclude that microwaveinduced hyperthermia can create retinal scarring without significant damage to sclera and choriocapillaris. The next experimental step will be to refine the microwave delivery system to ensure predictable and reproducible lesions.

Camu‐camu Myrciaria dubia (HBK) McVaugh: Chemical composition of fruit

AbstractAbstract: The chemical composition of the Peruvian camu‐camu fruit Myrciaria dubia(HBK) McVaugh was analysed at three stages of maturity (immature, midripe and ripe). As fruit matured, levels of ascorbic and dehydroascorbic acids, reducing sugars (fructose and glucose were the major sugars), amino acids (serine, valine and leucine) and soluble solids increased. Citric acid was the major acid (from 19.8 up to 29.8 g kg−1) and was responsible for the fruit's sour taste. Unlike citric acid, malic acid increased with maturation. Among the macronutrients, potassium was the most abundant mineral (711 mg kg−1) and could be considered, like vitamin C, nutritionally significant. During maturation, the fruit pulp colour turned from yellow‐green to pink, presumably due to the migration of anthocyanin pigments from the peel.

Behavioral selection in crayfish correlates with movement of a screening pigment in the eye

Light induces two contrasting behavioral responses in crayfish: attraction at low intensities and withdrawal at high intensities. The aim of our experiments has been to study whether screening pigments of the eye influence the selection of attraction or withdrawal responses. During illumination, screening pigments mask photoreceptor cells, reducing the gain of the visual system. Comparison of the time and light-intensity functions of pigment migration and of attraction and withdrawal responses suggest that pigment migration might influence the selection as well as the latency of the response.

The use of Q-switched Nd:Yag laser for removal of permanent eyeliner tattoo.

We present the results of a pilot study of the use of Nd:Yag laser in the removal of permanent eyeliner tattoos. Fourteen eyelids were treated, and in all cases a reduction in pigmentation was achieved although responses were variable. The advantages of such therapy over existing techniques are described. The technique offers a useful reduction of permanent eyeliner pigmentation and is particularly suited to treatment of localized areas of imperfect application or pigment migration.

Measurements of light transmission through single melanophores.

A photometrical method has been developed that allows assessment of subcellular pigment migration in melanophores of the fish cockoo wrasse (Labrus ossifagus L.) The pigment migration was studied with local light spot transmission measurements. Depending on where the light beam is placed on the melanophores it is possible to study events within an area of approximately 75 microns 2. Measuring pigment translocation in different parts of a melanophore gives new possibilities to study how cell membrane receptor-mediated signals are spread within a single cell, which will increase our understanding of how receptor activating drugs exert their cellular effect. The technique can be used in pharmacological and biophysical studies and in biosensors, pharmaceutical screens, environmental detectors, etc. The method clearly has the ability to study local and small changes in light transmission due to displacement of melanophore pigment granules. Since one melanophore on the tip of an optical fibre would be enough to obtain a measurable effect, the presented technique provides the basis for future development of biosensors small enough for in vivo applications, e.g., to monitor the catecholamine levels of circulating blood.

Long-term study of patients with congenital pit of the optic nerve and persistent macular detachment.

During the period 1970-87 we evaluated the changes of the optic disc, peripapillary area, detached macula and visual acuity in 16 cases with congenital pit of the optic nerve and macular detachment. The study revealed in 9 of the 16 cases (56%) an increase of the dimension of the pit or changes in its color, findings which were directly related to the duration of the macular detachment. Chorioretinal scarring, pigment migration, or both, were also noted mainly at the temporal margin of optic disc. In 5/16 cases we found during the follow-up an extension of macular elevation. In altogether 10 out of 16 cases the retinal elevation covered the larger portion of the mid-periphery temporally. In 7/16 cases the final visual acuity remained unchanged, in 9/16 cases deteriorated. The difference, however, in the latter 9 cases between initial and final visual acuity was negligible. During the follow-up period deterioration of the visual fields was also noted.

Conditional inhibition of screening-pigment aggregation by lidocaine in crayfish photoreceptors and frog retinal pigment epithelium.

Lidocaine, at concentrations equal to or lower than those that inhibit fast axoplasmic transport, was found to interfere with the dark-adapting migration of the screening pigments along crayfish photoreceptors and within the cells of the frog retinal pigment epithelium (RPE). The effects of the anesthetic on pigment movements were studied in isolated eyes incubated under light or dark conditions in media of different ionic compositions. Treatment of crayfish eyes with 25 mmol l-1 lidocaine in normal Van Harreveld's saline arrested pigment migration to the dark-adapted position or caused migration towards the light-adapted position in the dark. Similar results were obtained with frog eyecups exposed to 5 mmol l-1 lidocaine in Ringer's solution. In each case, the inhibition of dark adaptation was reversible and dependent on the levels of Na+ and Ca2+ in the incubation medium. A dark-adapted position of both pigments was compatible with lidocaine treatment provided that low-Na+, or high-Ca2+ or Co(2+)-containing solutions were used. These results indicate that light-adapted and dark-adapted pigment positions in both types of retinal cells can occur in the absence of local nervous input. Further, the data suggest a direct effect of lidocaine upon the photoreceptors or RPE cells. The inhibition of pigment aggregation is interpreted to be a consequence of an anesthetic-induced increase in the permeability of the plasma membrane, which in turn affects the intracellular ionic balance that controls pigment position.

Effects of steady electric fields on human retinal pigment epithelial cell orientation and migration in culture.

Low-level, steady electric fields of 6-10 volts/cm stimulated directional orientation and translocation of cultured human retinal pigment epithelial cells. The orientative movements (galvanotropism) consisted of somatic elongation of the cells into spindle shapes, followed by pivotal alignment orthogonal to the field. The anodal edges of the cells underwent retraction of their plasmalemmal extensions, while the cathode edges and the longitudinal ends developed lamellipodia and ruffled membranes. These tropic movements were followed by a translocational movement (galvanotaxis) of the cells towards the cathode. Staining of these migrating cells for actin showed the accumulation of stress fibers at the leading (cathodal) edge, as well as at the longitudinal ends of the elongated somata. These results suggest that endogenous, biologically-generated electric fields (eg., injury currents) may play a role in the guidance and migration of retinal pigment epithelial cells after retinal injury.

New Mechanism of Melanosomes Transport into Hair of Lambs of Different Breeds and Genotypes

By light microscopic investigation of skin and wool specimens of newborn lambs, we discovered a previously unknown mechanism for melanosomes transport in the process of dermal papilla melanocytes regular mitosis and migration into the hair shaft. This mechanism plays a great role in hair pigmentation especially in dominant (ED/ED) and recessive (Aa/Aa) black lambs of all investigated breeds. The rate of pigment cell mitosis, proliferation, and migration differs greatly in lambs of investigated color genotypes. In black genotypes the rate of melanocyte mitosis is very high and is approximately the same as in the hair bulb matrix cells, whereas in brown and red genotypes this rate is much lower. Melanocyte mitosis in the light red and tan groups was not found.

Funduscopic morphology of the light- and dark-adapted eye of the toadfish (Opsanus tau) in vivo

Intraocular funduscopy and photography permitted serial examination and documentation of time-related changes in the living toadfish (Opsanus tau) eye as it went from the dark-adapted to the light-adapted state. In the present study, the fully dark-adapted eye showed virtually no choroidal, pigment epithelial, or retinal vascular detail. The optic nerve was chalky white with indistinct borders. Grayish brown to chalky white retinal streaks radiated from the optic nerve into the periphery. As light adaptation began, the retinal streaks faded, with the retinal vasculature and the underlying choroidal and pigment epithelial detail becoming more discernible. Once the eye became fully light adapted, the typical geographic orange and black mottled appearance of the underlying pigment epithelium and choroid was visible, and the optic nerve head developed a pink hue with a distinct border. Even in the fully light-adapted eye, a vague radiation of lines could be seen extending from the optic nerve. This was superimposed on a cross-hatched mosaic of black dots seen throughout the fundus, not visible in the fully dark-adapted eye. Histological examination of the retina indicated forward migration of pigment in the pigment epithelium in the light-adapted state.

The ultrastructure of the compound eye of Munida rugosa (Crustacea: Anomura) and pigment migration during light and dark adaptation.

The galatheid squat lobster, Munida rugosa, has compound eyes of the reflecting superposition type in which a distal cone cell layer and a proximal rhabdom layer are separated by an extensive clear zone. The eye is shown to have certain unique features. In all other reflecting superposition eyes, the clear zone is traversed by crystalline tracts formed by the cone cells. In M. rugosa a thin distal rhabdom thread, formed by the eighth retinula cell, connects the cones to the proximal fusiform rhabdoms. The cytoplasm of the other retinula cells also crosses the clear zone in a complex pattern. Fully light-adapted ommatidia are optically isolated by limited migrations of distal shielding pigments. A reflecting pigment multilayer lines each cone to facilitate the formation of a superposition image. This also shows a light-induced change which may limit the acceptance angle of the eye during light adaptation.

Depth related variation in the structure and functioning of the compound eye of the Norway lobster Nephrops norvegicus

The mobility and quantity of retinula cell proximal screening pigment, and the liability of the eyes to light-induced damage, were investigated in the Norway lobster, Nephrops norvegicus (L.), obtained from three separate populations from depths of 18, 75, and 135 m.During the morning after capture, the migration of the proximal pigment in response to the onset of illumination below the threshold for damage varied between the three populations. In the eyes of deep water N. norvegicus, the proximal screening pigment was located close to or below the basement membrane when dark-adapted and rose to a position midway up the rhabdoms when light-adapted. In the dark-adapted N. norvegicus from shallow water the proximal pigment was located more distally than in eyes of deep water animals. After the onset of illumination, the pigment migrated distally to completely cover the rhabdoms. The amount of retinula cell proximal screening pigment was found to decrease linearly with depth.When dark-adapted individuals from each depth were exposed to light a positive correlation was obtained between the photon fluence rate (PER) and the proportion of the retina damaged. For a given light exposure the amount of damage was highest in animals from deeper water. The PFR causing 25% damage was approximately 1 log unit higher in animals from 18 m compared to those from 135 m.The amount of damage varied with the delay between capture of the animals and exposure to light. When exposed 2 h after capture significant differences between depths were seen but the results were influenced by the incomplete dark adaptation of some specimens.

Dual controls for screening pigment movement in photoreceptors of theLimuluslateral eye: Circadian efferent input and light

The radial and longitudinal distribution of retinular screening pigment in the lateral eye of the horseshoe crab Limulus polyphemus was quantified under a variety of experimental conditions. Pigment position was characterized by the center and width of the radial distribution at four levels in the ommatidium. Under diurnal lighting, intact animals show movement of pigment granules from the periphery of the retinular cell at night towards the junction of the arhabdomeral and rhabdomeral segments of the retinular cell in the day. In constant darkness, intact animals exhibit the same circadian rhythm in pigment migration. Animals with bilaterally cut optic nerves do not receive circadian efferent input from the brain and show little pigment movement in diurnal lighting. In all of these cases, pigment was either aggregated in a band just peripheral to the rays of the rhabdom or dispersed to the periphery of the retinular cell. When dark-adapted animals are exposed to a sudden large light increment, pigment moves inward between the rays of the rhabdom. During the day, this inward response begins immediately and reverses as the ommatidial aperture begins to close. At night, the onset of the inward movement is delayed, but then occurs more rapidly than during the day. No significant longitudinal movement of photoreceptor screening pigment was detected under any of these experimental conditions. Two opposing mechanisms control the movement of screening pigment in these cells. Release of neurotransmitters from the circadian efferents causes outward movement; large increments of light cause inward movement. In the absence of sudden changes in light intensity, circadian efferent input, not cyclic lighting, appears to be the major determinant of screening pigment position. A sudden and large increment of light triggers the rapid inward movement which appears to be a protective mechanism optimized for daytime performance.

Myosin and actin in melanophore-like variants of goldfish erythrophoroma cells: their intracellular distribution and possible association with pigment translocation

The occurrence and intracellular distribution of myosin and actin in melanophore-like cells derived from a goldfish erythrophoroma cell line have been studied by means of sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblot and immunofluorescence using antisera against chick gizzard myosin heavy chain and carp skeletal muscle actin. SDS-PAGE of the cell extracts separates out one band at 200 kDalton; this is conjugated with the anti-myosin antiserum. Immunofluorescence using the anti-myosin antiserum discloses that myosin in these cells occurs in two forms: discrete, minute clusters and thin filaments bearing a resemblance to “stress fibers”. The former is distributed evenly over the entire cytoplasm in the cells with dispersed pigments and, upon pigment aggregation, accumulates densely around collapsed melanosomes. The latter runs as thin bundles either radially along the cell center-to-periphery axis or connecting the corners of cell margins; it gives a similar profile in all states of the motile response. Immunofluorescence using the antiactin antiserum or rhodamine-conjugated phalloidin discloses that actin is similarly distributed to myosin, suggesting its possible existence as actomyosin. Simultaneous translocation of the amorphous forms of myosin and actin with melanosomes indicates that they may be involved in pigment migration.

A comparison of the α-adrenoceptor systems regulating pigment migration in erythrophores and melanophores of Labrus ossifagus

1. The regulation of pigment migration in erythrophores and melanophores of Labrus ossifagus was compared. 2. Both types of chromatophores are regulated by α 2-adrenoceptors, but the receptors of the erythrophores were more sensitive to adrenoceptor agonists than were those of melanophores. 3. The α 2-adrenoceptor of the erythrophores were prazosin-sensitive. 4. Forskolin (10μM) completely blocked pigment aggregation in both types of chromatophores. 5. Li + at a concentration of 100 μM interfered with pigment aggregation in erythrophores but not in melanophores. 6. The Na +/H + blocker amiloride inhibited pigment migration induced by the α 2-selective agonist B-HT 920 in both types of chromatophores. 7. There are differences in the α 2-adrenoceptor systems regulating pigment migration in erythrophores and melanophores of Labrus ossifagus.

Effect of Neurotransmitters on Movement of Screening Pigment in Insect Superposition Eyes

Abstract Com pound Eye, Pigment Migration, Adrenaline, Octopamine, Noradrenaline The light-induced expansion of the screening pigment in the com pound eyes of the sphingid moth Deilephila and the owlfly Ascalaphus is counteracted by local application of noradrenaline or octopamine. Ata critical concentration, similar for both drugs, the expanding effect caused by a light stimulus is completely neutralized, and at higher drug concentrations light stimulation induces a contraction of the screening pigment. The contracting effect of noradrenaline and octopamine is counteracted by adrenaline. None of the other putative neuroactive substances tested (acetylcholine, GABA , histamine, melatonin, serotonin and taurine) has a com parable effect on light-induced pigment movement. It is hypothesized that pigment migration is controlled by a system of antagonistically acting catecholamines, similar to the noradrenaline/adrenaline system present in vertebrates.

Eye Color Changes Due to Pigment Migration in Some Species of Heteroptera and Homoptera

Changes in eye color in response to light and darkness were observed in Trigonotylus doddi (Distant) (Heteroptera: Miridae), Saccharosydne saccharivora Westwood, Perkinsiella saccharicida Kirkaldy (Homoptera: Delphacidae), and Cyarda melichari Van Duzee (Homoptera: Flatidae). Histological examinations of the eyes of P. saccharicida revealed that pigments were condensed in a zone proximal to the crystalline cones when light-adapted, and dispersed distally into the region of the cones when dark-adapted. This adaptation is believed to occur primarily in nocturnal insects or species that are active by day and night. Eye color changes were not observed in 23 additional species of Heteroptera and Homoptera collected in Southern Florida, including Unerus colonus (Uhler) (Homoptera: Cicadellidae), which is reported for the first time from the United States.

Retinomotor movements in the frog retinal pigment epithelium: Dependence of pigment migration on Na + and Ca 2+

The ionic dependence of the screening-pigment migrations in the frog retinal epithelium (RPE) was quantitatively studied with eyecups incubated in media of different compositions. Typical migrations in response to light and darkness, equivalent to those observed in the intact animal, were fully accomplished and maintained for up to 6 hr by the isolated organ bathed in Ringer solution rich with O 2. Pigment migration in either direction was completed under the appropriate illumination conditions at any time during the day, indicating that circadian influences, if present in the intact animal, can be overridden in the isolated organ by light or darkness alone. Pigment aggregation toward the dark-adapted position was inhibited by: (a) low external Ca 2+, (b) high external Na +, and (c) drugs expected to increase the cytoplasmic levels of either Na +, or Ca 2+, like ouabain, caffeine and the ionophore A23187. However, the inhibition caused by low Ca 2+ did not occur if Na + was also reduced in the incubation medium. On the other hand, an increase in the concentration of external Ca 2+ or the addition of Co 2+ to the normal Ringer facilitated pigment aggregation in the dark. Pigment dispersion to the light-adapted position was unaffected by any of the above conditions. This is the first report of full and stable pigment responses in the RPE of vertebrate eyes incubated under simple physiological conditions. The results seem to conciliate a discrepancy of previous reports on the Ca 2+ dependence of RPE movements, and are compatible with current views on ionic mechanisms in analogous systems of intracellular transport.