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An animal’s perceived sense of orientation depends upon the head direction (HD) system found in several limbic structures and depends upon an intact peripheral vestibular labyrinth. However, how the vestibular system influences the generation, maintenance, and updating of the HD signal remains poorly understood. Anatomical and lesion studies point towards three key brainstem nuclei as being potential critical components in generating the HD signal: nucleus prepositus hypoglossi (NPH), supragenual nucleus (SGN), and dorsal paragigantocellularis reticular nuclei (PGRNd). Collectively, these nuclei are situated between the vestibular nuclei and the dorsal tegmental and lateral mammillary nuclei, which are thought to serve as the origin of the HD signal. To test this hypothesis, extracellular recordings were made in these areas while rats either freely foraged in a cylindrical environment or were restrained and rotated passively. During foraging, a large subset of cells in all three nuclei exhibited activity that correlated with changes in the rat’s angular head velocity (AHV). Two fundamental types of AHV cells were observed: 1) symmetrical AHV cells increased or decreased their neural firing with increases in AHV regardless of the direction of rotation; 2) asymmetrical AHV cells responded differentially to clockwise (CW) and counter-clockwise (CCW) head rotations. When rats were passively rotated, some AHV cells remained sensitive to AHV whereas others had attenuated firing. In addition, a large number of AHV cells were modulated by linear head velocity. These results indicate the types of information conveyed in the ascending vestibular pathways that are responsible for generating the HD signal.Significance StatementExtracellular recording of brainstem nuclei (nucleus prepositus hypoglossi, supragenual nucleus, and dorsal paragigantocellularis reticular nucleus) that project to the head direction circuit identified different types of angular head velocity (AHV) cells while rats freely foraged in a cylindrical environment. The firing of many cells was also modulated by linear velocity. When rats were restrained and passively rotated some cells remained sensitive to AHV, whereas others had attenuated firing. These brainstem nuclei provide critical information about the rotational movement of the rat’s head in the azimuthal plane.

Chemotaxis of sperm is an important step toward fertilization. During chemotaxis, sperm change their swimming behavior in a gradient of the chemoattractant that is released by the eggs, and finally sperm accumulate near the eggs. A well established model to study chemotaxis is the sea urchin Arbacia punctulata. Resact, the chemoattractant of Arbacia, is a peptide that binds to a receptor guanylyl cyclase. The signaling pathway underlying chemotaxis is still poorly understood. Stimulation of sperm with resact induces a variety of cellular events, including a rise in intracellular pH (pHi) and an influx of Ca2+; the Ca2+ entry is essential for the chemotactic behavior. Previous studies proposed that the influx of Ca2+ is initiated by the rise in pHi. According to this proposal, a cGMP-induced hyperpolarization activates a voltage-dependent Na+/H+ exchanger that expels H+ from the cell. Because some aspects of the proposed signaling pathway are inconsistent with recent results (Kaupp, U.B., J. Solzin, J.E. Brown, A. Helbig, V. Hagen, M. Beyermann, E. Hildebrand, and I. Weyand. 2003. Nat. Cell Biol. 5:109–117), we reexamined the role of protons in chemotaxis of sperm using kinetic measurements of the changes in pHi and intracellular Ca2+ concentration. We show that for physiological concentrations of resact (

Pyramidal cells in the rat hippocampus commonly show place-related activity, but it has been difficult to understand the factors that govern them. A particularly important question is whether individual cells have identifiable correlates that can be manipulated independently of the correlates of other cells. Recently Tanila et al. examined the activity of small ensembles of hippocampal cells in rats running on a plus-maze with distinct intra- and extramaze cues. When the two sets of cues were rotated 90° in opposite directions, some cells followed the intramaze cues, others followed the extramaze cues, and others “remapped” unpredictably; moreover, all possible combinations were seen within simultaneously recorded ensembles. In the current study, CA1 pyramidal cell population activity was recorded from four rats in a similar paradigm, using a recording system that permitted the analysis of ensembles of 4–70 simultaneously recorded units. The results were consistent with the data from the earlier study in showing an increase in remapping over time and in showing some place fields following one of the defined sets of cues while others remapped. When the possibility of random remapping was controlled for, however, the analysis did not show significant numbers of place fields following both sets of cues simultaneously. Furthermore, all rats initially showed fully concordant responses with all place fields following the local cues. For two rats, this pattern continued until a new configuration was introduced at which time all fields switched to follow the distal cues. Taken together, the results are difficult to reconcile with the hypothesis that individual hippocampal cells encode information about different subsets of cues in the environment.

Neural activity in several limbic areas varies as a function of the animal's head direction (HD) in the horizontal plane. Lesions of the vestibular periphery abolish this HD cell signal, suggesting an essential role for vestibular afference in HD signal generation. The organization of brain stem pathways conveying vestibular information to the HD circuit is poorly understood; however, recent anatomical work has identified the supragenual nucleus (SGN) as a putative relay. To test this hypothesis, we made lesions of the SGN in rats and screened for HD cells in the anterodorsal thalamus. In animals with complete bilateral lesions, the overall number of HD cells was significantly reduced relative to control animals. In animals with unilateral lesions of the SGN, directional activity was present, but the preferred firing directions of these cells were unstable and less influenced by the rotation of an environmental landmark. In addition, we found that preferred directions displayed large directional shifts when animals foraged for food in a darkened environment and when they were navigating from a familiar environment to a novel one, suggesting that the SGN plays a critical role in projecting essential self-motion (idiothetic) information to the HD cell circuit.

Successful navigation requires a constantly updated neural representation of directional heading, which is conveyed by head direction (HD) cells. The HD signal is predominantly controlled by visual landmarks, but when familiar landmarks are unavailable, self-motion cues are able to control the HD signal via path integration. Previous studies of the relationship between HD cell activity and path integration have been limited to two or more arenas located in the same room, a drawback for interpretation because the same visual cues may have been perceptible across arenas. To address this issue, we tested the relationship between HD cell activity and path integration by recording HD cells while rats navigated within a 14-unit T-maze and in a multiroom maze that consisted of unique arenas that were located in different rooms but connected by a passageway. In the 14-unit T-maze, the HD signal remained relatively stable between the start and goal boxes, with the preferred firing directions usually shifting

Heparin is thought to inhibit InsP3 binding to receptors involved in the intracellular release of Ca2+. Injection of heparin into Limulus ventral photoreceptors to high intracellular concentrations reduces the amplitude and slows the rate of rise of voltage-clamp currents induced by brief flashes, tends to make the responses to long flashes more "square," and tends to block the light-induced rise in [Ca2+]i detected by arsenazo III. In these ways, intracellular heparin mimics the effects of high concentrations of intracellular BAPTA or EGTA. In addition, the effects of heparin are attenuated by prior injection of BAPTA to high intracellular concentrations. Neomycin and spermine are thought to inhibit phospholipase C activity. Injections of spermine or neomycin to low intracellular concentrations largely mimic the effects of intracellular heparin. These findings suggest that the predominant effect of polyamines is to inhibit light-induced production of InsP3 by phospholipase C activity and thereby reduce the light-induced increase in [Ca2+]i. Our findings suggest that excitation can proceed in the absence of InsP3-induced increases in [Ca2+]i, but (a) the gain and speed of transduction are reduced and (b) adaptation is largely blocked.

Rapid concentration jumps of Ins(1,4,5)P3 or ATP were made inside Limulus ventral photoreceptors by flash photolysis of the parent caged compounds. In intact ventral photoreceptors, the photolysis flash evokes a maximum amplitude light-activated current; therefore, a procedure was developed for uncoupling phototransduction by blocking two of the initial reactions in the cascade, rhodopsin excitation and G protein activation. Rhodopsin was inactivated by exposure to hydroxylamine and bright light. This procedure abolished the early receptor potential and reduced the quantum efficiency by 325 +/- 90-fold (mean +/- SD). G protein activation was blocked by injection of guanosine-5'-O-(2-thiodiphosphate) (GDP beta S). GDP beta S injection reduced the quantum efficiency by 1,881 +/- 1,153-fold (mean +/- SD). Together hydroxylamine exposure and GDP beta S injection reduced the quantum efficiency by 870,000 +/- 650,000-fold (mean +/- SD). After the combined treatment, photoreceptors produced quantum bumps to light that was approximately 10(6) times brighter than the intensity that produced quantum bumps before treatment. Experiments were performed with caged compounds injected into photoreceptors in which phototransduction was largely uncoupled. Photolysis of one compound, myo-inositol 1,4,5-triphosphate P4(5)-1-(2-nitrophenyl)ethyl ester (caged IP3), increased the voltage clamp current in response to the flashlamp by more than twofold without changing the latency of the response. The effect was not seen with photolysis of either adenosine-5'-triphosphate P3-1-(2-nitrophenyl)ethyl ester (caged ATP) or caged IP3 in cells preloaded with either heparin or (1,2-bis-(o-amino-phenoxy)ethane-N-N-N'-N' tetraacetic acid tetrapotassium salt (BAPTA). The results suggest that photoreleased IP3 releases calcium ions from intracellular stores and the resulting increase in [Ca2+]i enhances the amplification of the phototransduction cascade.

Head direction (HD) cells in the rat anterodorsal thalamic nucleus (ADN) fire relative to the animal’s directional heading. Lesions of the entire vestibular labyrinth have previously been shown to severely alter VIIIth nerve input and disrupt these HD signals. To assess the specific contributions of the semicircular canals without altering tonic VIIIth nerve input, ADN cells were recorded from chinchillas following bilateral semicircular canal occlusion. While ADN HD cells (and also hippocampal place cells and theta cells) were identified in intact chinchillas, no direction-specific activity was seen following canal occlusions. Instead, “bursty” cells were observed that exhibited burst-firing patterns similar to normal HD cells, but with firing unrelated to the animal’s actual head direction. Importantly, when pairs of bursty cells were recorded, the temporal order of their firing was dependent upon the animal’s turning direction, as is the case for pairs of normal HD cells. These results suggest that bursty cells are actually disrupted HD cells. The present findings further suggest that the HD cell network is still able to generate spiking activity following canal occlusions, but the semicircular canal input is critical for updating the network activity in register with changes in the animal’s HD.

Aspects of a possible involvement of hydrogen ions in the electrophysiological responses to light of Limulus ventral photoreceptors were investigated. A I M solution of either a zwitter-ionic pH buffer or a weakly-buffering control substance was pressure injected through a micropipette into a ventral photoreceptor cell. To estimate the amount injected, 35SO4 was included in the solution. Membrane currents induced by light flashes were measured by a voltage-clamp technique. The buffer-filled micropipette passed current and a 3 M KCl filled micropipette monitored membrane voltage. The sensitivity (peak light-induced current/stimulus energy) was measured, after dark adaptation, before and after injection. Injections of buffers, pH 6.3–7.2, to intracellular concentrations of at least 40–200 mM produced only a small mean decrease in sensitivity, approximately equal to that caused by injections of control substances. Excitation, therefore, apparently is not mediated by a change in intracellular pH. Buffers with pH values 5.4–8.4 were also injected. The time to peak of the response depended on pH, being shortened by up to 20% at pH values below 7.7 and lengthened at higher pH values. The time to peak of the response appeared to be shortened by an increase in intracellular pH-buffering capacity even when there was no change in intracellular pH.

The spectral sensitivities of single Limulus median ocellus photoreceptors have been determined from records of receptor potentials obtained using intracellular microelectrodes. One class of receptors, called UV cells (ultraviolet cells), depolarizes to near-UV light and is maximally sensitive at 360 nm; a Dartnall template fits the spectral sensitivity curve. A second class of receptors, called visible cells, depolarizes to visible light; the spectral sensitivity curve is fit by a Dartnall template with λmax at 530 nm. Dark-adapted UV cells are about 2 log units more sensitive than dark-adapted visible cells. UV cells respond with a small hyperpolarization to visible light and the spectral sensitivity curve for this hyperpolarization peaks at 525–550 nm. Visible cells respond with a small hyperpolarization to UV light, and the spectral sensitivity curve for this response peaks at 350–375 nm. Rarely, a double-peaked (360 and 530 nm) spectral sensitivity curve is obtained; two photopigments are involved, as revealed by chromatic adaptation experiments. Thus there may be a small third class of receptor cells containing two photopigments.

A hyperpolarization can be recorded intracellularly following either a single bright light stimulus or the intracellular injection of Na(+). This after-hyperpolarization is abolished by bathing in 5 x 10(-6)M strophanthidin or removal of extracellular K(+). Both treatments also lead to a small, rapid depolarization of the dark-adapted cell. When either treatment is prolonged, light responses can still be elicited, although with repetitive stimuli the responses are slowly and progressively diminished in size. The rate of diminution is greater for higher values of [Ca(++)](out); with [Ca(++)](out) = 0.1 mM, there is almost no progressive diminution of repetitive responses produced by either K(+)-free seawater or strophanthidin. We propose that an electrogenic Na(+) pump contributes directly to dark-adapted membrane voltage and also generates the after-hyperpolarizations, but does not directly generate the receptor potential. Inhibition of this pump leads to intracellular accumulation of sodium ions, which in turn leads to an increase in intracellular Ca(++) (provided there is sufficient extracellular Ca(++)). This increase in intracellular calcium probably accounts for the progressive decrease in the size of the receptor potential seen when the pump is inhibited.

During intracellular iontophoretic injection of Ca(++) into Limulus ventral photoreceptor cells, there is a progressive diminution of the light response. Following Ca(++) injection, the size of the light response slowly recovers. Similarly, there is a progressive diminution of the light response during intracellular injection of Na(+) and recovery after the injection is stopped. The rate of diminution during Na(+) injection is greater for higher [Ca(++)](out). In solutions which contain 0.1 mM Ca(++), there is nearly no progressive decrease in the size of the light response during Na(+) injection. Intracellular injections of Li(+) or K(+) do not progressively decrease the size of the light response. We propose that an increase in [Na(+)](in) leads to an increase in [Ca(++)](in) and that an increase in [Ca(++)](in) by any means leads to a reduction in responsiveness to light.

Spectral sensitivity curves can be distorted by screening pigments. We have determined whether this is true for Limulus polyphemus by determining, from receptor potentials recorded using intracellular microelectrodes, spectral sensitivity curves for normal animals and for white-eyed animals (which lack screening pigment). Our results show: (a) In median ocelli, the curve for UV-sensitive receptor cells peaks at 360 nm and does not depend on the presence of screening pigment, (b) The curve for ventral eye photoreceptors is identical to that for retinular cells from the lateral eyes of white-eyed animals and peaks at 520–525 nm. (c) In normal lateral eyes, when the stimulating light passes through screening pigment, the curve indicates relatively more sensitivity in the red region of the spectrum than does the curve for white-eyed animals. Therefore, the screening pigment is probably red-transmitting, (d) In median ocelli, the curve for visible-sensitive cells peaks at 525 nm and is approximately the same whether the ocelli are from normal or white-eyed animals. However, the curve is significantly broader than that for ventral eyes and for lateral eyes from white-eyed animals.

The dark-adapted current-voltage (I-V) curve of a ventral photoreceptor cell of Limulus, measured by a voltage-clamp technique, has a high slope-resistance region more negative than resting voltage, a lower slope-resistance region between resting voltage and zero, and a negative slope-resistance region more positive than 0 v. With illumination, we find no unique voltage at which there is no light-induced current. At the termination of illumination, the I-V curve changes quickly, then recovers very slowly to a dark-adapted configuration. The voltage-clamp currents during and after illumination can be interpreted to arise from two separate processes. One process (fast) changes quickly with change in illumination, has a reversal potential at +20 mv, and has an I-V curve with positive slope resistance at all voltages. These properties are consistent with a light-induced change in membrane conductance to sodium ions. The other process (slow) changes slowly with changes in illumination, generates light-activated current at +20 mv, and has an I-V curve with a large region of negative slope resistance. The mechanism of this process cannot as yet be identified.

The receptor potential in Limulus photoreceptors appears to be a consequence not of permeability changes in the cell membrane but of alterations in a light-sensitive constant-current generator.

Membranes from ventral photoreceptors of Limuluswere incubated with cholera toxin and [32P]NAD+. Cholera toxin catalyzes a specific ADP-ribosylation of a 43-kDa peptide from Limulus ventral photoreceptors. Possible homologies between the 43-kDa peptide of Limulus and the α-subunits of mammalian stimulatory, guanine nucleotide-binding regulatory component of adenylate cyclase (Ns) were investigated by comparing the electrophoretic patterns of proteolytic fragments derived from each of these peptides that are radiolabeled by [32P]NAD+ and cholera toxin. Evidence is provided for structural homology between this invertebrate peptide and mammalian Ns.

There are two classes of photoreceptor cells in the median ocellus of Limulus. One class of cells respond to long wavelength (visible) stimuli with a depolarizing receptor potential and to near ultraviolet light with a biphasic, initially hyperpolarizing, receptor potential. The other class of receptors respond with a depolarization to near ultraviolet and with a biphasic response to visible light. In the latter type of cell, visible light can counteract the depolarization elicited by near ultraviolet light. The evidence suggests that there are two photopigments in each cell and that both are involved in the generation of receptor potential.

The membranes of photoreceptor cells in Limulus have an electrogenic sodium pump which contributes directly to membrane potential and whose activity is changed by light. These light-induced changes in pump activity underlie the receptor potential.

Two types of photoreceptors are found in the median ocellus of Limulus. One type is maximally sensitive to ultraviolet (UV) light, the other to green light; they are called UV and VIS cells, respectively. Biphasic receptor potentials, consisting of a small initial hyperpolarizing phase and a later slow depolarizing phase, can be recorded from both receptor types. These biphasic responses are elicited in UV cells in response to long-wavelength light, and in VIS cells in response to ultraviolet light. Another type of hyperpolarizing response can be recorded in UV cells: after a bright ultraviolet stimulus, the cell remains depolarized; long-wavelength light rapidly returns the membrane potential to its value preceding ultraviolet illumination (this long-wavelength-induced potential change is called a "repolarizing response"). Also, a long-wavelength stimulus superimposed during a UV stimulus elicits a sustained repolarizing response. A third cell type (arhabdomeric cell) found in the median ocellus generates large action potentials and is maximally sensitive to UV light. Biphasic responses and repolarizing responses also can be recorded from arhabdomeric cells. The retina is divided into groups of cells; both UV cells and VIS cells can occur in the same group. UV cells in the same group are electrically coupled to one another and to an arhabdomeric cell.

In the UV-sensitive photoreceptors of the median ocellus (UV cells), prolonged depolarizing afterpotentials are seen following a bright UV stimulus. These afterpotentials are abolished by long-wavelength light. During a bright UV stimulus, long-wavelength light elicits a sustained negative-going response. These responses to long-wavelength light are called repolarizing responses. The spectral sensitivity curve for the repolarizing responses peaks at 480 nm; it is the only spectral sensitivity curve for a median ocellus electrical response known to peak at 480 nm. The reversal potentials of the repolarizing response and the depolarizing receptor potential are the same, and change in the same way when the external sodium ion concentration is reduced. We propose that the generation of repolarizing responses involves a thermally stable intermediate of the UV-sensitive photopigment of UV cells.

Horizontal cells in the retinas of cats and rabbits are morphologically similar; in both species, two types can be distinguished in Golgistained material. Horizontal cells and their processes are readily recognized in electron micrographs, and many of the horizontal cell processes appear to make synaptic contacts with dendrites and somata of bipolar cells, and probably with other horizontal cells. The synapses of the horizontal cell appear similar to chemical synaptic contacts described throughout the nervous system. With the finding of synaptic contacts, it seems clear that retinal horizontal cells should be classified as neurons.