Lateral Eyes Research Articles

Circadian Structural Changes in the Retina of Lycosa tarentula (Araneae: Lycosidae)

Daily structural changes of the rhabdoms are described in the four eye types of a lycosid spider submitted to laboratory conditions: LD 12:12 (light on at 08.00, 100 lux). From a study of animals submitted to constant darkness during fourteen days, rhabdom turnover was shown to be circadian. Each type of retina exhibits its own rhythmic turnover of rhabdoms. Anterior-lateral, posterior-median and lateral eyes are mostly diurnal; anterior-median retinae comprise 300 receptors with mainly nocturnal functioning and 150 receptors with mainly diurnal functioning. A correlation was found between rhythms of locomotor activity, other activity, and rhabdom turnover in L. tarentula suggesting that the same pacemaker controls these different rhythms.

A Myosin III fromLimulusEyes Is a Clock-Regulated Phosphoprotein

The lateral eyes of the horseshoe crab Limulus polyphemus undergo dramatic daily changes in structure and function that lead to enhanced retinal sensitivity and responsiveness to light at night. These changes are controlled by a circadian neural input that alters photoreceptor and pigment cell shape, pigment migration, and phototransduction. Clock input to the eyes also regulates photomechanical movements within photoreceptors, including membrane shedding. The biochemical mechanisms underlying these diverse effects of the clock on the retina are unknown, but a major biochemical consequence of activating clock input to the eyes is a rise in the concentration of cAMP in photoreceptors and the phosphorylation of a 122 kDa visual system-specific protein. We have cloned and sequenced cDNA encoding the clock-regulated 122 kDa phosphoprotein and show here that it is a new member of the myosin III family. We report that Limulus myosin III is similar to other unconventional myosins in that it binds to calmodulin in the absence of Ca2+; it is novel in that it is phosphorylated within its myosin globular head, probably by cAMP-dependent protein kinase. The protein is present throughout the photoreceptor, including the region occupied by the photosensitive rhabdom. We propose that the phosphorylation of Limulus myosin III is involved in one or more of the structural and functional changes that occur in Limulus eyes in response to clock input.

Chromatographic isolation of PP2A from Limulus lateral eyes. Conventional and small scale methods.

Chromatographic isolation of PP2A from Limulus lateral eyes. Conventional and small scale methods.

A cGMP-gated cation channel in depolarizing photoreceptors of the lizard parietal eye.

Rods and cones of the two vertebrate lateral eyes hyperpolarize when illuminated, a response generated by a cyclic GMP cascade leading to cGMP hydrolysis and consequently the closure of cGMP-gated, non-selective cation channels that are open in darkness. Lizards and other lower vertebrates also have a parietal (third) eye, which contains ciliary photoreceptors that under dark-adapted conditions depolarize to light instead. Depolarizing light responses are characteristic of most invertebrate rhabdomeric photoreceptors, and are thought to involve a phosphoinositide signalling pathway (see, for example, refs 7-9). Surprisingly, we have found in excised membrane patches a cGMP-gated channel that is selectively present at high density on the outer segment (the presumptive light-sensitive part) of the parietal eye photoreceptor. Like the light-activated channel of the cell, it is non-selective among cations. Inositol trisphosphate (InsP3) had no effect on the same membrane patches. These findings suggest that the photoreceptors of the parietal eye, like rods and cones, use a cGMP cascade and not an InsP3-mediated pathway for phototransduction, but in this case light increases cGMP. A unifying principle of evolutionary significance emerges: that phototransductions in various ciliary photoreceptors, whether hyperpolarizing or depolarizing, uniformly use a cGMP cascade and a cGMP-gated channel to generate the light response, although there are rich variations in the details.

The effect of neuropeptides from Limulus on its circadian rhythm in retinal sensitivity.

Retinal responses of the Limulus lateral eyes to light are greater at night than during the day. A circadian clock in the brain of the horseshoe crab controls these rhythmic changes of light sensitivity. The increase in sensitivity (as measured by the amplitude of the electroretinogram) is mediated at least in part by octopamine that is released from efferent axons terminating in the visual cells. Earlier studies indicate that certain factors in Limulus hemolymph can act in conjunction with octopamine. More recently, five neuropeptides (LP1-LP4 and Lip-HP) had been isolated from acetone extracts of the Limulus central nervous system using HPLC fractionation and radioimmunoassay with antisera against FMRFamide-like peptides for detection. Presently, we have injected into the Limulus lateral eye these five peptides and observed changes in retinal sensitivity. Injection during daytime had no immediate effect on that daytime electroretinogram but decreased the electroretinogram amplitude for the entire subsequent night (12 h). However, upon injection at night, we observed an immediate but only transitory decrease in electroretinogram amplitude for about 1 h without effect on the subsequent daytime electroretinogram. We suggest that the peptides act antagonistically to octopamine and are highly dependent upon the activity state of the efferent nerve terminals.

Molecular and immunological characterization of a Gq protein from ventral and lateral eye of the horseshoe crabLimulus polyphemus

GTP binding proteins of the Gq family have been implicated in phototransduction in rhabdomeral photoreceptors. In this study we used molecular and immunochemical techniques to characterize a GTP-binding protein alpha subunit of the Gq family in ventral and lateral photoreceptors of the horseshoe crab, Limulus polyphemus. Both ventral photoreceptors and lateral eye retinular cells became strongly labeled with an antibody directed against the common carboxyl tail of two Gq family proteins, G alpha q and G alpha 11. This antibody also labeled a 42 kDa band on Western blots of proteins from ventral photoreceptor cell bodies, ventral photoreceptor axons, lateral eyes and lateral optic nerves. The reverse transcription-polymerase chain reaction (RT-PCR), along with degenerate oligonucleotide primers designed against conserved regions of known G alpha q and G alpha 11 proteins, was used to isolate a cDNA from ventral eye RNA which encodes a protein with high identity to known Gq proteins. Ribonuclease protection assays showed that the corresponding message was expressed in ventral eye, but these assays, as well as Northern blots, failed to detect expression in lateral eye. Therefore, while photoreceptors of both ventral and lateral eyes contain a Gq-like protein, the mRNA encoding the Gq protein in the ventral eye may differ in nucleotide sequence from its lateral eye counterpart.

Ovarian Follicular Kinetics in Response to Enucleation and Parietal Shielding inRana cyanophlyctis

Ovarian follicular kinetics, gravimetric changes in the ovary and the oviduct, were studied in the skipper frog,Rana cyanophlyctis,following enucleation (ENX) and parietal shielding (PS), during the breeding reason. PS was to prevent extraoptic photoreception by the pineal via the frontal organ. The gonadosomatic index (GSI) was high in ENX and PS frogs (P<0.01). The highest GSI occurred in frogs that were both ENX and PS (P<0.01). Percentage of oviductal weight decreased (P<0.01) in PS frogs and increased (P<0.01) in those subjected to both ENX and PS. Oviductal weight of ENX frogs was unchanged. Data on follicular kinetics revealed a decrease (P<0.01) in the ENX group in the number of first growth phase (FGP) oocytes and atretic follicles (AF), whereas the number of medium-sized secondary growth phase (MSGP) oocytes and large-sized secondary growth phase (LSGP) oocytes increased (P<0.01). There was a similar pattern of follicular kinetics in ENX+PS frogs. However, following PS, the number of FGP oocytes and AF increased (P<0.01) and decreased (P<0.01) in the MSGP and LSGP oocytes. The lower oviductal weight in PS frogs correlated with reduced SGP oocytes, the major source of estrogen. The importance of lateral eyes and the frontal organs in the transduction of environmental signals to the neuroendocrine–reproductive axis is indicated in this species. That recruitment of FGP to SGP is inhibited only in the PS group suggests that the frontal organ plays an important role in ovarian follicular kinetics.

Receiver operating characteristic (ROC) analysis of neural code efficacies. I. Graded photoreceptor potentials and data quality.

Nerve cell signals are different in form from the stimuli that evoke them and they exhibit complex spatio-temporal characteristics. This defines a neural coding problem which is addressed by two current theories: Multiple Meaning Theory holds that neural signals contain patterns that make statements about combinations of stimulus properties; the Task Dependence Hypothesis suggests that different features of identical neural signals mediate performance in different behavioral tasks. These coding issues were addressed by investigating the representation of sensory information in the distal nervous system after transduction of visual stimuli into bio-electric signals. The objects of study were light-evoked neural responses which had been intracellularly recorded from single retinula (photoreceptor) cells in Limulus lateral eyes. The efficacies with which sensory information was represented by various candidate neural codes were calculated using receiver operating characteristic (ROC) analyses to provide objective indices. The specific visual problem under investigation was discrimination between light flashes whose intensities differed by a very small amount. A wide range of light adaptation states and relative stimulus intensities were explored. Extremely stringent data quality standards were applied which restricted the investigation to cells whose potentials did not exhibit any statistically significant drift during the hours required for data collection. Seven cellular characterizations were simultaneously monitored to detect drift in a given cell's potentials; these characterizations included the value of the membrane potential and the values of six candidate codes. These codes were: the area under the light-evoked receptor potential (RP), the mean value of the RP, the peak height of the RP, the slope of the onset of the RP, the duration required for the RP to drop from its peak by a given amount, and the duration required for the RP to end. The results were: (1) Light adaptation increases efficacy. (2) Thus, light adaptation trades sensitivity for acuity (as characterized by ROC discriminations). (3) Increasing relative light flash intensity also increases efficacy. (4) The efficacies of the various codes are significantly different and fall in the following order: area > or = peak = mean > or = duration-end = slope = duration-drop. These findings further demonstrate that arbitrary characterizations of stimulus-response relationships are very likely to be incomplete. They particularly indicate that many commonly used and quite conventional neural analysis strategies may substantially under-estimate system performance.

Receiver operating characteristic (ROC) analysis of neural code efficacies. II. Optic nerve action potentials and neural transmission.

Action potentials (APs) were intracellularly recorded from eccentric cells (which give rise to optic nerve fibers) in Limulus lateral eyes and their neural coding efficacies were determined over a wide range of light adaptation states and relative stimulus intensities. Extremely stringent data quality procedures were followed to ensure that the results are based on stable preparations. Waveforms which could be compared with those of comparable receptor potentials (RPs) were obtained by constructing plots of the reciprocals of successive interspike intervals, creating instantaneous spike frequency waveforms (ISFWs). Six candidate codes were then measured. They were: the area under the light-evoked ISFW, the mean value of the ISFW, the peak height of the ISFW, the slope of the onset of the ISFW, the duration required for the ISFW to drop from its peak by a given amount, and the duration required for the ISFW to end. Receiver operating characteristic (ROC) analyses were then applied to these coded ISFW values to provide objective indices of AP efficacies in the form of detectability (d') measurements. Several reliable findings were obtained: (1) Both adaptation state and relative intensity affect efficacy. More specifically, the d' values of all codes approach zero, representing chance detectability, when relatively weak flashes are presented in dark-adapted states. (2) Light adaptation produces a sensitivity-acuity tradeoff: as sensitivity decreases in more light-adapted states, detectabilities increase, indicating that ROC-characterized discrimination acuity increases. (3) The AP code efficacy order is similar to but not identical with the efficacy order previously found in photoreceptors: area = peak > or = mean > or = slope = duration-end = duration-drop. The previously measured photoreceptor RP efficacies were quantitatively compared with the present AP data from the optic nerve fiber level, with the following results: (1) All efficacies tend to decline at the more proximal neural level. (2) The decline is code dependent and transmission efficacy falls in this order: peak > area > mean > slope = duration-end = duration-drop. (3) The code which transmits best between photoreceptors and optic nerve fibers (i.e., the peak) differs from the code which has the highest efficacy at the photoreceptor level (i.e., the area); at the more proximal level, these two codes have indistinguishable efficacies. These findings support two conclusions: (1) They further demonstrate that arbitrary characterizations of stimulus-response relationships are very likely to be incomplete. This would be particularly important when, as is often the case in brain research, the mean code alone (i.e., the average firing rate) is used to characterize spike potentials. The present data show that the use of that code would have substantially underestimated detectability had it been used alone. (2) The fact that the code which most completely transmits information between cells is not the code which most completely represents information within a cell leads directly to a possible physiological basis for the existence of multiple neural codes. It particularly leads to the following extension of the Task Dependence Hypothesis of neural coding: Different codes may mediate performance in different behavioral tasks because different codes best serve different neural functions.

Spectral Sensitivity of Single Photoreceptor Cells in the Eyes of the Ctenid Spicier Cupiennius salei Keys

Intracellular recordings from photoreceptor cells of all eyes of the spider Cupiennius salei Keys reveal 3 groups of cells with spectral sensitivity maxima in the blue (480 nm). green (520 nm), and UV (340 nm), respectively. The blue and green cells show secondary peaks in the UV at 340-380 nm. In the posterior median, posterior lateral and anterior median eyes, the majority of photoreceptor cells are the blue and green cells in roughly equal numbers. In the anterior lateral eye, however, the green cells dominate. UV cells were only found in the secondary eyes and only once in each of them.

Characterization of protein phosphatases type 1 and type 2A in Limulus nervous tissue: their light regulation in the lateral eye and evidence of involvement in the photoresponse.

The activities of both protein phosphatases and protein kinases are responsible for the transient changes in the levels of phosphorylation and probably the functions of protein intermediates involved in the biochemical and physiological mechanisms underlying the photoresponse in photoreceptor cells from both vertebrate and invertebrate organisms. Of the known protein serine/threonine phosphatases, various forms of type 1 (PP 1) and type 2A (PP 2A) protein phosphatases are responsible for dephosphorylating many of the known phosphoproteins including those involved in photoreceptor cell function. In this report, we provide biochemical evidence for both PP 1- and PP 2A-like activities in the visual and nonvisual tissue of the horseshoe crab, Limulus polyphemus, that membrane and soluble forms of both enzymes are present, and that the activities of both enzymes are greater in light- than in dark-adapted lateral eyes. These activities were characterized using glycogen phosphorylase a, a substrate for both PP 1 and PP 2A, and various protein phosphatase inhibitors, including okadaic acid. We also report that okadaic acid, at concentrations required to inhibit PP 1, inhibited physiological functions of photoreceptor cells from the ventral eye, causing a delayed reduction of the resting membrane, and slowing and reducing light responses.

The Ocular Anomaly in a Ground Spider(Araneae: Gnaphosidae) from Japan.

The ocular anomaly in a Japanese ground spider is reported. The spider is identified with Gnaphosa primorica OVTSHARENKO, PLATNICK et SONG, 1992, described from the Russian Far East. It is recorded for the first time in Japan. The anomalous individual has only two eyes; these are regarded as the right anterior lateral and posterior lateral eyes.

Efferent Control in the Anterior Lateral Eyes of Orb Weaving Spiders

Abstract The anterior lateral (AL) eyes of the orb weaving spiders, Argiope amoena and A. bruennichii, have a single type of receptor cell with a maximum sensitivity at about 520–540 nm. Efferent optic nerve signals control the photoreceptor response in the AL eyes. Specifically, efferent signals decrease the threshold for light responses and change the waveform of responses. Similar changes in the threshold and waveform could be produced by exogenous application of octopamine, suggesting that it is an efferent neurotransmitter.

The morphology and physiology of a "mini-ommatidium" in the median optic nerve of Limulus polyphemus.

Examination of the Limulus median optic nerve with low-magnification light microscopy allows clear visualization of an ultraviolet-sensitive mini-ommatidium enshrouded by pigment cells, glial cells, and guanophores. Serial 1-micron sections of median optic nerves containing mini-ommatidia revealed the presence of a single, heavily pigmented photoreceptor (retinular) cell and a single, unpigmented arhabdomeric cell. Computer-assisted serial reconstructions from 1-micron sections confirmed the presence of two cells, each bearing a nucleus, and two axons leaving the mini-ommatidium. The retinular cell is morphologically similar to retinular cells from the median and lateral eyes. Its rhabdomere appears to be a continuous sheet of microvilli with much infolding. The structure of the arhabdomeric cell is nearly identical to those found in the median ocellus. As in other photoreceptors in Limulus, the retinular cell of the mini-ommatidium is innervated by efferent fibers from the brain. Each mini-ommatidium generates a single train of nerve impulses in response to light, presumably from the arhabdomeric cell. Measurement of the spectral sensitivity of the mini-ommatidium based upon a constant-response criterion indicated that the retinular cell is maximally sensitive to near ultraviolet light with lambda max = 380 nm. Comparison of intensity-response functions revealed that those of the mini-ommatidium are significantly steeper than those of the ocellus almost certainly as the result of neural processing in the ocellus which is absent in the mini-ommatidium.

Isolation and Expression of an Arrestin cDNA from the Horseshoe Crab Lateral Eye

Electrophysiological studies of photoreceptors from the horseshoe crab Limulus polyphemus continue to provide fundamental new knowledge of the photoresponse in invertebrates. Therefore, it is of particular interest to characterize the molecular components of the photoresponse in this system. Here we describe an arrestin cloned from a cDNA library constructed using poly(A)+ RNA isolated from Limulus lateral eyes. The protein, deduced from the arrestin cDNA, is most similar to arrestin from locust antennae (56% identity) and Drosophila phosrestin I (53% identity). Limulus arrestin was expressed in a heterologous system, and its properties were compared with those of a 46-kDa light-regulated phosphoprotein (pp46A) in Limulus photoreceptors described in previous studies from this laboratory. Arrestin and pp46A (a) have the same apparent molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (b) have an isoelectric point in the basic pH range, (c) require calmodulin and elevated Ca2+ levels for phosphorylation, (d) are immunoreactive with monoclonal antibody C10C10 directed against a sequence in bovine arrestin (S-antigen) that is perfectly conserved in the deduced arrestin protein, and (e) are associated with photoreceptors. We conclude that the arrestin described here and pp46A are the same protein. The results of this and previous studies show that in Limulus photoreceptors, light regulates the phosphorylation of arrestin in complex ways.

Opsins from the lateral eyes and ocelli of the horseshoe crab, Limulus polyphemus.

cDNA clones encoding opsins from the lateral eyes and median ocelli of the horseshoe crab, Limulus polyphemus, were isolated from cDNA libraries. The opsin cDNAs obtained from the lateral eye and ocellar libraries code for deduced proteins with 376 amino acids. The two cDNAs are 96% identical at the nucleic acid level, differing primarily at the 3' untranslated region, and are apparently the products of two separate genes. The deduced opsin proteins are 99% identical to each other, differing at only 5 amino acids. The opsins encoded by these cDNAs are most likely the protein moiety of the visible-wavelength rhodopsins in this animal. In the lateral eye, expression of the opsin gene is restricted to the photoreceptor cells of the ommatidia. Comparisons with opsins of other species show that the Limulus opsin proteins are most similar (53% identity) to the opsin from the R1-6 photoreceptors of flies.

Antagonistic chromatic mechanisms in photoreceptors of the parietal eye of lizards.

Photoreceptors are the first in the chain of neurons that process visual information. In lateral eyes of vertebrates, light hyperpolarizes rod and cone photoreceptors that synapse onto bipolar and horizontal cells in the first synaptic layer of the retina. The sign of the photoreceptor signal is either conserved or inverted in bipolar cells, resulting in chromatically dependent depolarizing and hyperpolarizing responses to visual stimuli. Visual information is then conveyed to the second synaptic layer for encoding and transmission to the brain by ganglion cells. The parietal (third) eye of lizards does not contain bipolar cells or other interneurons. Photoreceptors synapse directly onto ganglion cells and yet, even in the absence of interneurons, antagonistic chromatic mechanisms modulate the ganglion cell responses. We report here that chromatic antagonism in the third eye originates in the chromatically dependent hyperpolarizing and depolarizing response of the photoreceptors to light. We also suggest that the antagonistic nature of these photoresponses may provide lizards with a mechanism for the enhanced detection of dawn and dusk.

Role of dermal photoreceptors and lateral eyes in initiation and orientation of locomotion in lamprey

The response to illumination, and the functional roles of skin photoreceptors and lateral eyes, were examined in the adult river lamprey ( Lampetra fluviatilis L.). Illumination of one side of the lamprey evoked a turning movement away from the light source followed by locomotion. The lateral eyes were responsible for directing the movements away from the source of light. A selective illumination of one lateral eye consistently evoked a negative phototactic reaction, whereas a selective illumination of tail skin photoreceptors evoked locomotion, without any preferential orientation relative to the source of light. Experiments were performed by video recording the locomotor responses to localized illumination, and analyzed frame by frame. The horizontal turning movement during negative phototaxis consisted of an asymmetric laterally directed mechanical wave, of higher amplitude and lower velocity than the normal locomotory waves, which was propagated from the rostral to the caudal end of the body.

FUNCTIONAL MORPHOLOGY AND BEHAVIOR OF VARGULA HILGENDORFII (OSTRACODA: MYODOCOPIDA) FROM JAPAN, AND DISCUSSION OF ITS CRUSTACEAN ECTOPARASITES: PRELIMINARY RESULTS FROM VIDEO RECORDINGS

ABSTRACT Newly investigated (video recordings) aspects of the biology of myodocopid ostracodes are presented, exemplified by Vargula hilgendorfii (Muller, 1890) (Cypridinidae) from shallow marine environments of the Pacific coast of central Japan. Like other myodocopid species attracted to bait (Cohen, 1982; Cohen and Morin, 1986), Vargula hilgendorfii is infaunal by day and becomes active by night, either on the substratum (e.g., feeding activities on fish carcasses) or in the water column (e.g., vertical migrations) with associated blue luminescence (Irie, 1953). All individuals from the first juvenile (A-5) to the adult stage (both sexes) bore a yellow luminescent organ (upper lip region) and were able to emit light in laboratory conditions. A synchronized stroking movement of the second pair of antennae (A2) was responsible for various locomotory activities such as swimming and digging the sediment (bioclastic sand). The first antennae (A1) have a probable tactile function in the recognition of the substratum and also in copulatory activities (suckerlike features of males). The furcae facilitated the penetration into the sediment and assisted rotations in a vertical plane, on or within the substratum. Males with a stronger muscular protopodite and a smaller body were faster swimmers than females. Specimens almost invariably reacted to light by shifting and adjusting their moving direction to reach the least illuminated area of the experimental container and/or by digging the sediment. Diggings were superficial, rarely exceeding a few mm below the sediment-water interface; they are interpreted as a response (camouflaged shelter) against predation, most probably controlled and initiated by visual stimuli. The seventh limbs are complex transformed appendages used by the brooding female to stir her free embryos within the marsupium; the limbs were more commonly used by both sexes from the fourth juvenile stage (A-2) to brush any region of the domiciliar concavity (appendages, lateral eyes, vestment, body) and also part of the external margins of the valves. The morphology of the seventh limb (e.g., articulated stem, bell-shaped setae) and its peculiar oscillating movement, both contributed to eject foreign particles from the domicilium. The ancestry of the seventh limbs in myodocopid ostracodes can be traced to, at least, the Lower Triassic (Weitschat, 1983a). Smaller crustacean ectoparasites frequently inhabit Vargula hilgendorfii. Onisocryptus ovalis (Shiino, 1942) (Isopoda, Epicaridea) preferentially infects nonovigerous females or females with eggs in ovaries and clutches the body of its host near the heart. There is no evidence that this parasite feeds by tearing into the ostracode's integument. The cryptic habit of O. ovalis may reduce predatorial risks.

A circadian clock in the Limulus brain transmits synchronous efferent signals to all eyes.

A circadian clock in the brain of the horseshoe crab, Limulus polyphemus, has an important role in the function of the peripheral visual system. At night, the clock transmits neural activity to the lateral, ventral, and median eyes via efferent optic nerve fibers. The activity occurs in synchronous bursts (maximum rate of 2 bursts/s) with individual efferent fibers contributing a single spike in each burst. The circadian efferent activity originates in the protocerebrum. Lateral connections synchronize the efferent activity recorded from the two halves of the protocerebrum, suggesting the existence of bilateral circadian oscillators. Circadian efferent activity survives excision of the brain and isolation of the protocerebrum. We conclude that circadian clock and its complex neural circuitry are fundamental components of the Limulus visual system.