Ommatidial type-specific interphotoreceptor connections in the lamina of the swallowtail butterfly, Papilio xuthus

The eye of the butterfly Papilio xuthus contains a random array of three types of ommatidia (types I-III), each bearing nine photoreceptors, R1-R9. Of the six spectral classes of photoreceptors identified, types I, II, and III ommatidia contain four, three, and two classes, respectively: the ommatidia are thus spectrally heterogeneous. The photoreceptors send their axons to the lamina where, together with some large monopolar cells (LMCs), the nine from a single ommatidium contribute to a module called a lamina cartridge. We recently reported that among different photoreceptor axon terminals visualized by confocal microscopy, the number and length of axon collaterals differ for different spectral receptors, suggesting that lamina circuits are specific for each ommatidial type. Here we studied the distribution of synapse-like structures in the cartridges, first characterizing a photoreceptor by measuring its spectral sensitivity and then injecting Lucifer yellow (LY). We subsequently histologically identified the type of ommatidium to which the injected photoreceptor belonged, cut serial ultrathin sections of the entire lamina, labeled these with anti-LY immunocytochemistry, and then localized synapse-like structures. We found numerous interphotoreceptor contacts both within and between cartridges, the combination of which was again specific for the ommatidial type. R3 and R4, which are green-sensitive photoreceptors in all ommatidia, have thick axons lacking collaterals. We found that these cells exclusively make contacts with LMCs and not with photoreceptors. We therefore presume that R3 and R4 construct a system for motion vision, whereas other randomly distributed spectral types provide inputs for color vision.     

Photoreceptor projection reveals heterogeneity of lamina cartridges in the visual system of the Japanese yellow swallowtail butterfly, Papilio xuthus

The compound eye of the butterfly Papilio xuthus is composed of three types of spectrally heterogeneous ommatidia. The ommatidia, which contain nine photoreceptor cells, R1-9, bear four (type I), three (type II), or two (type III) classes of spectral receptors in fixed combinations. The photoreceptors send their axons to the lamina, the first optic ganglion, where the R1-9 axons originating from a single ommatidium, together with some second-order neurons, form a neuronal bundle, called a lamina cartridge. We investigated the axonal structure of photoreceptors in the lamina to determine whether the cartridge structure is different between the three ommatidial types. We first characterized a photoreceptor by measuring its spectral sensitivity and then injected Lucifer Yellow. We subsequently identified the type of ommatidium of the injected photoreceptor via histological sections. We further observed the axonal structure of the photoreceptor in the lamina by laser confocal microscopy. We found that the number and length of axon collaterals markedly differ between the spectral receptors. Those having the most extensive axon collaterals, which extend into six or more surrounding cartridges, are violet receptors (R1 and R2 of type II ommatidia). UV receptors (R1 or R2 of type I ommatidia) also send collaterals into two to four neighboring cartridges. Blue receptors (R1 or R2 of type I ommatidia, R1 and R2 of type III ommatidia) have short collaterals restricted to their own cartridges. We thus conclude that the neuronal circuit of the lamina cartridge differs between the three types of ommatidia.     

Mitochondria are redistributed in Drosophila photoreceptors lacking milton,a kinesin-associated protein

Photoreceptors are richly supplied with mitochondria, where they are required to meet the energetic demands, in the soma, of phototransduction and, in the terminal, of neurotransmitter release. Compromising the latter, we have made photoreceptors R1-R6 in Drosophila ommatidia homozygous for either of two alleles, milt(186) and milt(92), of milton in whole-eye mosaics. Such mutant photoreceptors fail to target mitochondria to their terminals. We show from quantitative electron microscopy (EM) that mitochondria are totally lacking at the terminal but nevertheless abundant and present throughout the soma, where their distribution differs from that of control ommatidia, however, being more heavily concentrated in the nuclear region. Mitochondria are sparse at the basalmost level of mutant ommatidia, and are lacking beneath the basement membrane, in the axons and terminals of these cells. The absence of mitochondria from R1-R6 terminals and concommitant reductions in synaptic vesicle packing density, previously reported, we show here are accompanied by reduced immunoreactivity to the photoreceptor transmitter histamine but not by any change in total head histamine content, as determined by high-performance liquid chromatography. Mutant terminals also contain vesicle profiles with a wider range of sizes. These two phenotypes suggest that the reduced availability of ATP when mutant terminals lack a mitochondrial supply compromises their ability to pump histamine into synaptic vesicles and perturbs membrane distribution within the terminal. In addition, a band of somata in the lamina cortex, at least some of which are postsynaptic neurons not homozygous for milton, also shows altered mitochondrial targeting, with abnormal clusters of mitochondria, as visualized by immunolabeling with anti-hsp and by serial EM. Within the lamina, terminals of mutant photoreceptors are penetrated by neighboring cells with invaginations that frequently contain mitochondria, suggesting that a mechanism exists for intercellular metabolic support. Our findings indicate the direct and compensatory responses in a population of neurons when mitochondria are not correctly targeted to their synaptic terminals.     

Neuropeptide modulation of photosensitivity. II. Physiological and anatomical effects of substance P on the lateral eye of Limulus

A system of efferent substance P-like immunoreactive fibers innervates the ommatidia of the Limulus lateral eye. Thus, we tested the physiological effects of substance P on the lateral eye by measuring the electroretinogram, a population potential reflecting the photoreceptors' response to light, under different experimental conditions. Substance P had no direct effect on the photoreceptors, but it induced an increase in their responsiveness to test flashes of light. The latency, magnitude, and duration of this reversible modulatory effect was dose-dependent. The lateral eye displays an endogenous circadian rhythm in its responsiveness to light. Application of exogenous substance P in the daytime causes an immediate rise as well as an increase in the nocturnal peak, while injection of one of its antagonists (D-Pro2, D-Phe7, D-Trp9 substance P) in the afternoon retards the normal rise in sensitivity and reduces the nighttime levels. Passive incubation with substance P antibodies at midnight caused a drop to diurnal levels of photosensitivity. Short-term changes in photosensitivity, similar in their nature to the substance P-induced ones, were caused by arousing the subjects. Arousal had an effect on the ongoing circadian rhythm similar to that of substance P application. Thus, the substance P efferent system may regulate neural responsiveness in both a short-term, environmentally induced manner, as well as for level setting in a circadian fashion. The mechanism for substance P-induced increases in photosensitivity involves changes in ommatidial structure: contraction of distal pigment cells, resulting in an increased aperture, and contraction of the retinular cells and rhabdom, resulting in a wider diameter of the latter. These structural modifications result in a greater angle of acceptance and increased light quantum catch.     

Molecular and genetic analysis of Drosophila eye development: sevenless, bride of sevenless and rough

Environmental cues play a critical role in neuronal development in the Drosophila compound eye. Through cell-cell interactions, different classes of photoreceptor neurons are generated in specific numbers and occupy characteristic positions within ommatidia, the building blocks of the compound eye. Recent studies on three genes, sevenless, bride of sevenless and rough, provide insight into the cellular rules and molecular mechanisms underlying ommatidial assembly.     

Visuomotor adaptation during inactivation of the dentate nucleus.

Recent experiments have suggested that the process of visuomotor adaptation depends on how a visual distortion is introduced. The cerebellum is thought to be involved in adapting to rapidly introduced visual distortions; however its role in adapting to a gradually introduced distortion is unknown. We tested adaptation to a sudden or a gradual introduction of a visual distortion, during reversible inactivation of a monkey's dentate nucleus. There was significant adaptation in both of these tasks without any lignocaine infusion and during saline infusions. However after inactivation the ability to adapt to either visual distortion was slightly impaired. This dysfunction was significant when the visuomotor distortion was introduced over several trials, suggesting that the cerebellum has a differential contribution to visual adaptation depending on the type of visuo-motor disturbance encountered.     

The retina and retinal projection on the lamina ganglionaris of the crayfish Pacifastacus leniusculus (Dana).

The retinular cell morphology and ommatidia arrangement in the crayfish retina are studied. Each ommatidium contains eight receptor cells (R1-R8). Seven of them (R1-R7) contribute to a large spindle-shaped rhabdom with orthogonal layers of microvilli. Distal to the rhabdom of R1-R7, the 8th receptor cell forms a separate rhabdom with horizontal microvilli. The cell is four-lobed, is devoid of screening pigments, and forms a thin axon projecting past the first optic neuropile (lamina ganglionaris) and has a terminal in the second (medulla externa), indicating a separate function of the receptor. The axons from the eight receptors of one ommatidium project to different synaptic compartments (cartridges) in the lamina. A pattern is described where eight axons from three adjacent ommatidia join in one cartridge; and conversely, the axons from one ommatidium split and join to three cartridges. The seven axons from R1-R7 terminate in two levels of the lamina plexiform layer (epl 1 and epl 2), four in the distal and three in the proximal part. Among the monopolar ganglion cells, two types are found with lateral branches restricted to either of the two receptor terminal layers (M3 in epl 1 and M4 in epl 2) and axons terminating in the second optic neuropile. A correlation between the two orthogonal channels for e-vector discrimination and the two levels of terminals within the lamina is suggested. The retina is divided into dorsal and a ventral part with a mirror symmetry axis horizontally in the eye.     

Responses of caudate nucleus neurons to different visual stimuli in the actively awake cat

The responses of caudate neurons to diffuse or local (a slit) light stimuli were studied extracellularly in alert cats during natural fixation of the gaze. The number of neurons responding to local light stimulation was much higher than that responding to diffuse light. Local light stimulation produced more extensive inhibition of neuronal activity as well as "off" responses. Some neurons responded selectively to the orientation of slits. All neuronal responses were recorded with latencies of 40-90 ms. The participation of the caudate nucleus in the analysis of visual sensory information is discussed.     

Comparative characteristics of response of neurons of the caudate nucleus to different sensory stimuli in awake cats

Extracellular responses of caudate neurons to light slits of various orientations and to clicks have been studied in awake cats. Neurons with specific reaction to particular orientation of the slit could change the response pattern when clicks were applied. The latencies of visual and auditory responses of the same caudate neurons could also differ significantly. Most of such cells were distributed in the body of the caudate nucleus. The possibility of participation of caudate neurons in analysis of visual information is discussed.     

Patterned rhodopsin expression in R7 photoreceptors of mosquito retina: Implications for species‐specific behavior

Visual perception of the environment plays an important role in many mosquito behaviors. Characterization of the cellular and molecular components of mosquito vision will provide a basis for understanding these behaviors. A unique feature of the R7 photoreceptors in Aedes aegypti and Anopheles gambiae is the extreme apical projection of their rhabdomeric membrane. We show here that the compound eye of both mosquitoes is divided into specific regions based on nonoverlapping expression of specific rhodopsins in these R7 cells. The R7 cells of the upper dorsal region of both mosquitoes express a long wavelength op2 rhodopsin family member. The lower dorsal hemisphere and upper ventral hemisphere of both mosquitoes express the UV‐sensitive op8 rhodopsin. At the lower boundary of this second region, the R7 cells again express the op2 family rhodopsin. In Ae. aegypti, this third region is a horizontal stripe of one to three rows of ommatidia, and op8 is expressed in a fourth region in the lower ventral hemisphere. However, in An. gambiae the op2 family member expression is expanded throughout the lower region in the ventral hemisphere. The overall conserved ommatidial organization and R7 retinal patterning show these two species retain similar visual capabilities. However, the differences within the ventral domain may facilitate species‐specific visual behaviors. J. Comp. Neurol. 516:334–342, 2009. © 2009 Wiley‐Liss, Inc.     

Calcium and the mechanism of light adaptation in vertebrate photoreceptors.

Vertebrate photoreceptors transduce the absorption of light into a hyperpolarizing change in membrane potential. The mechanism of transduction is becoming fairly well understood and has been shown to occur via a G protein-coupled decrease in cyclic GMP. Attention is now turning to the way the enzymatic machinery in the outer segment of the photoreceptor cell is modulated during light adaptation. Recent studies show that light adaptation cannot occur if changes in the concentration of cytoplasmic free calcium in the outer segment are prevented, suggesting that calcium functions as a second messenger in sensitivity regulation.     

Functional anatomy of the fiddler crab compound eye (Uca vomeris: Ocypodidae,Brachyura, Decapoda)

We describe the structural organization of the ommatidium in the compound eye of the fiddler crab, Uca vomeris, at both the light‐ and the electron‐microscopy levels. We pay particular attention to the organization of the optical system, the retinular cells, the rhabdom, and of pigment cells. Although the fiddler crab compound eye is of the apposition type, typical for Brachyuran crabs, we identify a number of novel, functionally relevant aspects of ommatidial organization that have not previously been described. The flat corneal facet lenses provide the main focusing power and therefore must contain a gradient of refractive index. Each ommatidium has the typical set of eight retinular cells, with a distal retinular cell R8 lying close to the proximal tip of the crystalline cone. R8 is shaped into four lobes, which are separated by proximal extensions of the four crystalline cone cells and of distal extensions of retinular cells R1–R7. The microvilli in the R8 rhabdom are not aligned in a uniform direction, while the microvilli of the main rhabdom show the typical crustacean pattern of alternating bands of horizontally (R3, R4, R7) and vertically aligned microvilli (R1, R2, R5, R6). We describe in detail the distribution and structural properties of screening pigment granules in the two types of pigment cells and in the retinular cells in the equatorial eye. We discuss the functional significance of this fine‐structural organization of the fiddler crab compound eye in relation to visual processing and visual ecology. J. Comp. Neurol. 522:1264–1283, 2014. © 2013 Wiley Periodicals, Inc.     

Neural organization of afferent pathways from the stomatopod compound eye

Crustaceans and insects share many similarities of brain organization suggesting that their common ancestor possessed some components of those shared features. Stomatopods (mantis shrimps) are basal eumalacostracan crustaceans famous for their elaborate visual system, the most complex of which possesses 12 types of color photoreceptors and the ability to detect both linearly and circularly polarized light. Here, using a palette of histological methods we describe neurons and their neuropils most immediately associated with the stomatopod retina. We first provide a general overview of the major neuropil structures in the eyestalks lateral protocerebrum, with respect to the optical pathways originating from the six rows of specialized ommatidia in the stomatopod's eye, termed the midband. We then focus on the structure and neuronal types of the lamina, the first optic neuropil in the stomatopod visual system. Using Golgi impregnations to resolve single neurons we identify cells in different parts of the lamina corresponding to the three different regions of the stomatopod eye (midband and the upper and lower eye halves). While the optic cartridges relating to the spectral and polarization sensitive midband ommatidia show some specializations not found in the lamina serving the upper and lower eye halves, the general morphology of the midband lamina reflects cell types elsewhere in the lamina and cell types described for other species of Eumalacostraca.     

Protein kinase C activators inhibit the visual cascade in Limulus ventral photoreceptors at an early stage.

The phosphoinositide cascade mediates visual transduction in invertebrate photoreceptors. Phospholipase C (PLC) catalyzes the hydrolysis of phosphatidylinositol bisphosphate, producing inositol trisphosphate (InsP(3)) and diacylglycerol (DAG). Protein kinase C (PKC) is a major target of DAG in many cell types. We have used PKC activators to investigate the function of the kinase in the phototransduction cascade in Limulus polyphemus ventral photoreceptors. Extracellular application of (-)-indolactam V (0. 03-30 microM) or phorbol-12,13-dibutyrate (10 microM) reversibly reduced the sensitivity of the electrical response of the photoreceptors to light by up to 1000-fold. The inert stereoisomer (+)-indolactam V and 4alpha-phorbol had no effect. The effect of (-)-indolactam V was antagonized by the PKC inhibitors bisindolylmaleimide I and G? 6976. Coapplication of bisindolylmaleimide V, used as a negative control compound for PKC inhibition, did not reduce the effectiveness of (-)-indolactam V. These findings are consistent with (-)-indolactam V activating PKC and desensitizing the light response. Furthermore, our pharmacological results indicate that PKC activation does not appear to play a role in light adaptation. We localized the position of the target of PKC in the visual cascade. We chemically excited the cascade at various stages to determine the kinase's target. PKC activation by (-)-indolactam V decreased the light-induced elevation of intracellular calcium but had no effect on the photoreceptor's excitatory response to intracellular injection of InsP(3). However, the PKC activator greatly reduced the excitation caused by GTP-gamma-S injection. We propose that PKC inhibits the visual transduction cascade at the G-protein and/or PLC stage.     

Mechanotransducing ion channels in C6 glioma cells

Mechanotransducing (MS) ion channels and images of the patch membrane were studied in cell-attached patches in C6 glioma cells. MS channel density was ∼0.08 to 0.5 channels/μm², channel conductance was ∼40 pS (at -40 mV), and the reversal potential was +15 mV. Replacement of NaCl with KCl, CsCl, or Na gluconate in the pipette solution was without substantial effect on the current-voltage relationship. Replacement of NaCl with NMDG (N-Methyl-D-Glucamine) Cl or reducing NaCl decreased the amplitude of inward currents at negative membrane potentials and caused the reversal potential to shift in the negative direction. Rapid application of suction to the back of the pipette usually elicited a fast (<0.1 s) appearance of channel activity. The peak (phasic) in channel activity was followed by a decrease to a constant (tonic) level of activity. The reduction in channel activity—called adaptation—was reduced at depolarizing membrane potentials and disappeared if too much pressure was applied. Positive pressure caused the patch membrane to curve toward the pipette tip, move in the direction of the tip, and evoke MS channel activity. Removal of the positive pressure caused the patch to move back to the original position. Conversely, negative pressure caused the patch membrane to curve away from the pipette tip, move away from the tip, and elicit MS channel activity. Gigohm seal resistances were always maintained during translational movement of the patch membrane. Tonic MS channel activity was not associated with translational movements of the patch membrane. Phasic and tonic channel activity were independent of the sign of curvature of the patch membrane. C6 glioma cells have rapidly adapting voltage-dependent MS ion channels, which are non-selective for monovalent cations, and belong to the stretch-activating class of mechanosensory ion channels. Adaptation in MS channels may allow the cell to limit the influx of cations in response to mechanical input. The selective loss of adaptation suggests that the MS channel's gate receives input from two sources. A minimal viscoelastic mechanical model of adaptation and two alternative models for translational movement of the patch are presented. © 1996 Wiley-Liss, Inc.     

Temperature dependency of light adaptation in bullfrog cone photoreceptors

Isolated, superfused bullfrog retinas were stimulated in such a manner as to allow examination of the dynamics of the shift in the stimulus-response curve that accompanies light adaptation of cone photoreceptors. Observed phenomena were highly temperature dependent with complex Arrhenius relationships, suggesting that the mechanisms responsible for the shift involve multiple enzyme systems.     

Properties of depolarizing bipolar cell responses to central illumination in salamander retinal slices.

The voltage and current responses of depolarizing bipolar cells to central illumination were studied by means of whole-cell recording in retinal slices of the larval tiger salamander, Ambystoma tigrinum. To stabilize the responses, it was necessary to limit exchanges between the cytosol and the solution in the patch pipette by reducing the diameter of the pipette tip opening. The current-voltage relationship of the cell membrane in darkness displayed a strong outward rectification, and the inward current evoked by light could be consistently reversed by depolarization only when tetraethylammonium was added to the pipette solution. As a result of the membrane non-linearity, increases in the intensity of bright lights caused relatively smaller amplitude increases in the voltage than in the current responses and the latter had a proportionally smaller after-effect. With larger pipette tip openings, the cytosol equilibrated with the pipette filling solution. Under these conditions the light-evoked responses gradually became slower and acquired an on-off pattern, their final amplitude and polarity being determined by the ratio of the chloride concentrations on each side of the cell membrane. This finding is interpreted as revealing the existence of two response components: a chloride-dependent on-off increase in conductance and a faster depolarizing input that was lost through diffusional exchange. Addition of GTP and ATP to the electrode filling solution had a stabilizing effect on the labile component, whether or not cyclic GMP was also included. Observations on the magnitude of the conductance changes and on diphasic reversals indicate that the labile response component, presumably representing direct input from photoreceptors, is caused by an increase in conductance. The resulting inward current was still present at a low intracellular chloride concentration and may be assumed, therefore, to be carried by a cation influx.     

Effect of chronic lithium on sensitivity to light in male and female bipolar patients

1. Sensitivity to white light was quantified in euthymic bipolar male and female patients maintained on long term lithium therapy and age and sex matched unmedicated controls. 2. The Dark Adaptation Threshold procedure was used to assess sensitivity of both the cone and rod photoreceptors to short pulses of light. 3. Male and female controls did not differ in sensitivity to light. 4. Male patients in comparison to both controls and female patients evidenced reduced sensitivity to light during the cone and rod portion of the dark adaptation procedure. 5. Female patients did not differ from controls on sensitivity to light. 6. Using these and other published data the results were interpreted as suggesting that lithium reduces sensitivity to light during adaptation to dark.     

The origin of the Drosophila subretinal pigment layer

Optical insulation plays a critical role in the fine visual acuity of the Drosophila compound eye. Screening pigments expressed by a number of cell types contribute to this phenomenon. They provide optical insulation that prevents extraneous light rays from inappropriately activating the photoreceptors. This optical insulation can be divided into two categories; the insulation of the individual ommatidia, and the insulation of the compound eye as a whole. The whole-eye insulation is provided by two sources. The sides of the eye are optically insulated by the pigment rim, a band of pigment cells that circumscribes the eye. The base of the eye is insulated by the subretinal pigment layer; a thick layer of pigment that lies directly underneath the retina. How this subretinal pigment layer is generated has not been clearly described. Here, experiments that manipulate pigment expression during eye development suggest that the subretinal pigment layer is directly derived from pigment cells in the overlying retina.