Activity at night in efferent nerve fibers from a central circadian clock produces changes in photoreceptor function in the lateral compound eye of Limulus: the response to light is increased; membrane potential fluctuations (bumps) occurring in the dark are suppressed; and the duration of bumps occurring both in the dark and under dim illumination is increased (Barlow et al., 1977; Kaplan & Barlow, 1980; Barlow, 1983; Barlow et al., 1985, 1987). Efferent nerve terminals release octopamine when activated (Battelle et al., 1982; Battelle & Evans, 1984, 1986); exogenous octopamine in vitro produces some of the changes resulting from efferent nerve activity in vivo (Kass et al., 1988).

We report here that the increase in both on-transient and steady-state response to light induced by octopamine in the lateral eye in vitro are concentration dependent with threshold at or below 100 nM, saturation at or above 100 µM, and half-maximal increase in the range 1–10 µM. Octopamine also reduces bump activity in the dark in a concentration-dependent way. Unlike the increase in light response, the dependence of this effect on octopamine concentration is extremely variable from specimen to specimen. The effects of exogenous octopamine on light response and bump activity can sometimes be reversed by removing octopamine from the medium bathing the in vitro preparation. Octopamine also increases bump duration, apparently in a concentration-dependent manner. We have not succeeded in reversing this increase in bump duration.

The concentration dependence of changes in photoreceptor response described here agrees qualitatively with the dependence of cAMP levels on octopamine in Limulus photoreceptors (Kaupp et al., 1982), lending further support to the idea that cAMP acts as a second messenger in the circadian control of photoreceptor function. Our results also suggest that the changes induced in the transient and steady-state response to light by both efferent nerve activity and exogenous octopamine have a common origin, which may differ from that responsible for the modulation of bump activity.

The protein intermediates of the photoresponse and the modulation of this response in invertebrate photoreceptors are largely unknown. As a first step toward identifying these proteins, we have examined light-stimulated changes in protein phosphorylation in preparations of Limulus photoreceptors. Here we show that light modulates the level of phosphorylation of three proteins associated with Limulus ventral photoreceptors: the upper band of a 46-kD protein doublet (46A) and a 122-kD protein, which become more heavily phosphorylated in response to light, and the lower component of the 46-kD doublet (46B), which is phosphorylated in dark-adapted cells, but not in cells maintained in the light. In dark-adapted preparations, 46A is phosphorylated within 30 s after a flash of light and dephosphorylates over a period of many minutes. It is also a major substrate for calcium/calmodulin-dependent protein kinase (Wiebe et al., 1989); therefore, we speculate that 46A is involved in some aspect of dark adaptation. Interestingly, the level of phosphorylation of 46A is the same when measured from preparations maintained in complete darkness or ambient light for at least 1.5 h. The 122-kD phosphoprotein is the same protein which becomes phosphorylated in response to efferent innervation to Limulus eyes (Edwards et al., 1988) and the efferent neurotransmitter, octopamine (Edwards and Battelle, 1987). It may be involved in the increase in retinal sensitivity and the enhanced response of photoreceptors to light that is initiated by efferent innervation. Its role in light-stimulated processes is not clear. The level of phosphorylation of 46B may be most relevant to the long-term state of adaptation of the photoreceptor cell to light and dark.

Calcium (Ca2+) is thought to play a major role in the photoresponse of both vertebrates and invertebrates, but the mechanisms through which Ca2+ exerts its effects are unclear. In many systems, some effects of Ca2+ on cellular processes are thought to be mediated via activation of calcium/calmodulin protein kinase (CaCAM-PK) and the phosphorylation of specific proteins. Thus, protein substrates for CaCAM-PK in photoreceptor cells may be important in mediating the effects of Ca2+ on the photoresponse.

In this study, we identify eight substrates for CaCAM-PK found in both the ventral and lateral eyes of Limulus. We focus on a characterization of one of these, a 46-kD substrate. We show that its subcellular distribution in ventral photoreceptors and its isoelectric forms are identical to the 46-kD light-stimulated phosphoprotein (46A) described by Edwards et al. (1989). Furthermore, we present evidence that 46A is unique to photoreceptor cells, and that it is present throughout the cell. Based on the results of this study, and the previous study by Edwards et al. (1989), we propose that 46A is involved in mediating the effects of Ca2+ on Limulus photoreceptor cell function, and that it may be involved in dark adaptation.

The color vision of three domestic dogs was examined in a series of behavioral discrimination experiments. Measurements of increment-threshold spectral sensitivity functions and direct tests of color matching indicate that the dog retina contains two classes of cone photopigment. These two pigments are computed to have spectral peaks of about 429 nm and 555 nm. The results of the color vision tests are all consistent with the conclusion that dogs have dichromatic color vision.

Neurons in the nucleus praetectalis olivaris (NPO) were antidromically identified by electrical stimulation of the nucleus of Edinger-Westphal (EW), the location of preganglionic pupilloconstrictor motoneurons. Electrical stimulation within the NPO leads to bilateral pupil constriction. Single neurons recorded in the NPO respond tonically to light stimuli, and their discharge frequency increases linearly with logarithmic increase in light intensity. This characteristic identifies NPO neurons as luminance detectors. They have large receptive fields mostly lying in the upper and contralateral quadrant of the visual field.

Cats with impaired binocular vision show a significantly reduced binocular summation of the pupillary light reflex (BSP), i.e. the increase of pupil constriction during binocular illumination when compared to monocular illumination is less than in normal animals. The investigation of ocular dominance and subthreshold binocular interactions in the NPO of normal and innate microstrabismic cats revealed two possible mechanisms for BSP and its reduction in strabismic subjects. First, the percentage of neurons increasing their discharge rate by illuminating either eye is significantly reduced in the NPO of innate microstrabismic cats (6.6%) when compared to normal cats (22% of all neurons tested). Second, in most NPO neurons of normal cats the subthreshold influence of the ipsilateral eye leads to an increase in neuronal activity during binocular stimulation when compared to monocular stimulation of the contralateral eye (binocular summation). The subthreshold influence of the ipsilateral eye in most NPO neurons of microstrabismic cats, however, is inhibitory, i.e. the neuronal discharge rate during binocular stimulation is decreased when compared to monocular stimulation of the contralateral eye (binocular inhibition). However, there is no significant correlation between BSP and binocularity in the NPO in individual animals. This suggests that BSP may be additionally influenced by visual structures other than NPO.

The anatomical substrate of the pupillary light reflex was investigated in normal and innate microstrabismic cats using anatomical methods as well as electrical stimulation. The bilateral retinal input to the nucleus praetectalis olivaris (NPO), the pretectal relay station in the subcortical pupilloconstrictor pathway, was identified to come from the ventral retina where the upper visual field is represented. Orthodromic electrical stimulation revealed that retinal information is transmitted to on-tonic neurons in the NPO mainly via slowly conducting axons probably originating from W- and X-type retinal ganglion cells.

For the first time, a direct cortical input to on-tonic neurons in the NPO could be demonstrated. This cortical input originates from caudolateral parts of the occipital cortex. Putative input structures are those subdivisions of areas 19 and 20a where the upper part of the visual field is represented.

A direct, predominantly contralateral projection with a weak ipsilateral component from NPO to the nucleus of Edinger-Westphal, and an interhemispheric connection between the NPOs could be demonstrated. With respect to the anatomical connections as described in this study, no differences between normal and innate microstrabismic cats could be found.

The results are discussed with respect to the binocular summation of the pupillary light reflex and its reduction in subjects with impaired binocular vision.

In the present study, the anterograde tracer Phaseolus vulgaris-leucoagglutinin was injected into area V1 in order to demonstrate the detailed morphology of individual axons terminating in prestriate area MT. On the basis of 24 axon reconstructions, several representative (but not necessarily comprehensive) characteristics have been identified: (1) Most axons arborize in a patchy manner over a widespread territory, frequently greater than 1.0 mm and often up to 1.5 × 1.8 mm (dimensions uncorrected for shrinkage). (2) Terminal arbors are distributed to layers 3, 4, and 6. Those in layer 6 need not be in register with those in the upper layers. (3) Number and size of terminal arbors are variable. One axon may have 1–4 arbors in the middle layers; typically at least one of these will have a diameter of 200–250 µm, while the others may be less developed. There are from 1–3 arbors in layer 6, usually 50 µm (but sometimes up to 100 µm) in diameter. (4) Terminal boutons are of mixed morphology, but usually beaded and large (up to 3.0 µm). (5) In the white matter, many axons travel in the external sagittal stratum but some are part of the U-fiber system. Axons commonly branch, sometimes at depths up to 0.75–1.0 mm, below the gray matter of MT. In summary, these axons are not stereotyped, but rather vary in the number and size of their terminal arbors, as well as in their branching and overall geometry.

Connections from area V1 to MT have been associated with the magnocellular-dominated “processing channel.” As widespread arborizations and bistratified terminations are common to both striate axons in MT and to geniculocortical axons in layer 4Cα of primary visual cortex, these features might be correlated with magnocellular-specific processing requirements.

Inferior frontal eye field (FEF) projections to the dorsolateral pontine nucleus (DLPN), and corticocortical connections with the superior temporal sulcal (STS) cortex, were studied in five macaque monkeys which had received horseradish peroxidase (HRP) gel implants into the inferior prearcuate cortex (including area 45 of Walker, 1940). These connections were contrasted with those from the dorsal FEF (area 8a) in another macaque monkey. Findings of heavy inferior FEF projections to the ipsilateral DLPN (light to the contralateral DLPN) and reciprocal connections with the deep caudal bank and fundus of the superior temporal sulcus (STS), presumed to be the middle temporal (MT) visual area (Maunsell & Van Essen, 1983a), appeared to go hand in hand with more pronounced projections to the stratum superficialis of the superior colliculus (SC). In contrast, the HRP gel implant in the dorsal prearcuate cortex (area 8a of Walker, 1940) resulted in only very light projections to the ipsilateral DLPN, more pronounced projections to the dorsomedial pontine nucleus (DMPN), almost no projection to the stratum superficialis (SS), and more pronounced reciprocal connections with the upper bank of the STS, presumed to be the medial superior temporal (MST) area (Maunsell & Van Essen, 1983a). Both the inferior and dorsal FEF also had extensive reciprocal connections with the ventral intraparietal area (VIP; Maunsell & Van Essen, 1983a) in the caudal bank of the intraparietal sulcus. The correlated projections of the inferior FEF to the DLPN, MT area, and SS may explain its reported role in smooth pursuit (Lynch, 1987), in addition to its well-established role in the production of voluntary purposeful saccadic eye movements (Bruce et al., 1985).