Michiyo Kinoshita

Associative visual learning, color discrimination, and chromatic adaptation in the harnessed honeybee Apis mellifera L.

We studied associative visual learning in harnessed honeybees trained with monochromatic lights associated with a reward of sucrose solution delivered to the antennae and proboscis, to elicit the proboscis extension reflex (PER). We demonstrated five properties of visual learning under these conditions. First, antennae deprivation significantly increased visual acquisition, suggesting that sensory input from the antennae interferes with visual learning. Second, covering the compound eyes with silver paste significantly decreased visual acquisition, while covering the ocelli did not. Third, there was no significant difference in the visual acquisition between nurse bees, guard bees, and foragers. Fourth, bees conditioned with a 540-nm light stimulus exhibited light-induced PER with a 618-nm, but not with a 439-nm light stimulus. Finally, bees conditioned with a 540-nm light stimulus exhibited PER immediately after the 439-nm light was turned off, suggesting that the bees reacted to an afterimage induced by prior adaptation to the 439-nm light that might be similar to the 540-nm light.

Coexpression of two visual pigments in a photoreceptor causes an abnormally broad spectral sensitivity in the eye of the butterfly Papilio xuthus.

The compound eye of the butterfly Papilio xuthus consists of three different types of ommatidia, each containing nine photoreceptor cells (R1–R9). We have found previously that the R5–R8 photoreceptors of type II ommatidia coexpress two different mRNAs, encoding opsins of green- and orange-red-absorbing visual pigments (Kitamoto et al., 1998). Do these cells contain two functionally distinct visual pigments? First, we identified the sensitivity spectrum of these photoreceptors by using combined intracellular recording and dye injection. We thus found that the R5–R8 of type II ommatidia have a characteristic sensitivity spectrum extending over an excessively broad spectral range, from the violet to the red region; the photoreceptors are therefore termed broadband photoreceptors. The spectral shape was interpreted with a computational model for type II ommatidia, containing a UV visual pigment in cells R1 and R2, two green visual pigments in cells R3 and R4, a far-UV-absorbing screening pigment (3-hydroxyretinol) in the distal part of the ommatidium, and a red-screening pigment that surrounds the rhabdom. The modeling suggests that both visual pigments in the R5–R8 photoreceptors participate in phototransduction. This work provides the first compelling evidence that multiple visual pigments participate in phototransduction in single invertebrate photoreceptors.

An ultraviolet absorbing pigment causes a narrow-band violet receptor and a single-peaked green receptor in the eye of the butterfly Papilio

The distal photoreceptors in the tiered retina of Papilio exhibit different spectral sensitivities. There are at least two types of short-wavelength sensitive receptors: an ultraviolet receptor with a normal spectral shape and a violet receptor with a very narrow spectral bandwidth. Furthermore, a blue receptor, a double-peaked green receptor and a single-peaked green receptor exist. The violet receptor and single-peaked green receptor are only found in ommatidia that fluoresce under ultraviolet illumination. About 28% of the ommatidia in the ventral half of the retina exhibit the UV-induced fluorescence. The fluorescence originates from an ultraviolet-absorbing pigment, located in the most distal 70 microns of the ommatidium, that acts as an absorption filter, both for a UV visual pigment, causing the narrow spectral sensitivity of the violet receptor, and for a green visual pigment, causing a single-peaked green receptor.

Tuning of Photoreceptor Spectral Sensitivities by Red and Yellow Pigments in the Butterfly Papilio xuthus

Abstract The compound eye of the Japanese yellow swallowtail butterfly, Papilio xuthus, consists of different types of ommatidia characterized by the pigmentation around the rhabdom. About 75% of the ommatidia harbor red pigment, whereas the other 25% contain yellow pigment. We find that the pigments function as spectral filters for the proximal photoreceptor cells. Intracellular recordings of the proximal cells yielded spectral sensitivities peaking in the red (λmax = 600 nm) and in the green (λmax = 520 nm), respectively. Staining of the recorded cells and subsequent histology demonstrated that the red receptors contain red pigment and that the green receptors contain yellow pigment. The sensitivity spectrum of the red receptors was considerably narrower compared to the absorption spectrum of a visual pigment peaking at 600 nm. The sensitivity spectrum can be calculated with an optical model for the butterfly rhabdom by incorporating measured absorbance spectra of the red pigments, yielding that the cell contains a visual pigment peaking at about 575 nm. The model also indicated that the spectral sensitivity of the green receptors is produced by the combination of the yellow lateral filter and a visual pigment peaking at 515 nm.

Retinal regionalization and heterogeneity of butterfly eyes

The regional characteristics of the eyes of butterflies from different families have been surveyed using epi-illumination microscopy, utilizing the eyeshine visible due to the tapetum situated proximally to the rhabdom. All butterflies studied have a high spatial acuity in the frontal region. The facet diameter varies slightly across the eye, and the interommatidial angle and the eye parameter p are especially large dorsally. Whereas the ommatidial lattice is generally highly regular, the eyeshine colours distinctly depend on the species. Sometimes the eyeshine is locally uniform, but often it is heterogeneous. It is hypothesized that the regional characteristics as well as the local heterogeneity are adaptations that optimize spectral discrimination.

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. J. Comp. Neurol. 483:341–350, 2005.

Blue and Double-peaked Green Receptors Depend on Ommatidial Type in the Eye of the Japanese Yellow Swallowtail Papilio xuthus

Abstract The compound eye of the butterfly Papilio xuthus is composed of three spectrally distinct types of ommatidia. We investigated the blue and double-peaked green receptors that are encountered distally in type I and III ommatidia, by means of intracellular recordings, in vivo fluorescence microscopy, and histology. The blue receptors are R1 and/or R2 photoreceptors; they contain the same mRNA encoding the opsin of the blue-absorbing visual pigment. However, here we found that the sensitivity in the UV wavelength region strongly depends on the ommatidial type; the blue receptors in type I ommatidia have a distinctly depressed UV sensitivity, which is attributed to lateral filtering in the fused rhabdom. In the main, fronto-ventral part of the eye, the R3 and R4 photoreceptors of all ommatidia contain the same set of two mRNAs encoding the opsins of green-absorbing visual pigments, PxL1 and PxL2. The spectral sensitivities are double-peaked, but the UV sensitivity of the R3 and R4 photoreceptors in type I ommatidia appears to be reduced, similar to that of the co-localized blue receptors.