Effects of α2-Adrenergic Agonists and Antagonists on Photoreceptor Membrane Currents

Type B photoreceptors of the nudibranch mollusc Hermissenda crassicornis receive excitatory synaptic potentials (EPSPs) whose frequency is controlled by potential changes of a neighboring cell known as the S optic ganglion cell which is thought to be electrically coupled to the presynaptic source of these EPSPs, the E optic ganglion cell. The frequency of the EPSPs increases when a conditioned stimulus (light) is paired with an unconditioned stimulus (rotation) during acquisition of a Pavlovian conditioned response. The results of the present study are consistent with an adrenergic origin for these EPSPs. Noradrenergic agonists (greater than 100 microM), norepinephrine and clonidine, only slightly depolarize the type B cell but clearly prolong its depolarizing response to light. Serotonin, by contrast, causes hyperpolarization of the type B cell's resting potential as well as after a light step. Clonidine reduces voltage-dependent outward K+ currents (IA, an early current, ICa2+-K+, a late Ca2+-dependent current) that control the type B cell's excitability (and thus its light response and membrane potential). These effects of clonidine are reduced or blocked by the alpha 2-receptor antagonist, yohimbine (0.5 microM), but not the alpha 1-blocker, prazosin. The same yohimbine concentration also blocked depolarizing synaptic excitation of the type B cell in response to depolarization of a simultaneously impaled S optic ganglion cell. Histochemical techniques (both the glyoxylic acid method of de la Torre and Surgeon and the formaldehyde-induced fluorescence or Falck-Hillarp method) demonstrated the presence of a biogenic amine(s) within a single neuron in each optic ganglion as well as three or four cells within the vicinity of previously identified visual interneurons. No serotonergic neurons were found within the optic ganglion or in proximity to visual interneurons. A clonidine-like synaptic effect on type B cells, therefore, could amplify conditioning-specific changes of membrane currents by increasing type B depolarization and possibly, as well, by elevating intracellular second messengers.     

Lanthanum mimicks the trp photoreceptor mutant of Drosophila in the blowfly Calliphora

The effect of lanthanum on the light response of blowfly (Calliphora erythrocephala) photoreceptors was studied. The electrophysiological behaviour of the photoreceptors in the presence of La can be summarized as follows: 1. Upon long stimulation the photoreceptors responded with a 'transient receptor potential', i.e. the cells depolarized at the onset of the stimulus and then repolarized to (or below) the resting potential. This effect was dependent on stimulus intensity and occurred only at high intensities. During illumination membrane noise was reduced. 2. The light-induced changes in membrane potential were paralleled by changes in membrane resistance. 3. The time course of the receptor response was slowed down. 4. Light adaptation led to an increase in response latency. 5. The recovery of the receptor response after light adaptation was slowed down. 6. The sensitivity of the receptor cells measured by the response to short light stimuli was reduced. In summary, the electrophysiological behaviour of Calliphora photoreceptors in the presence of La was very similar to that of the photoreceptors of the trp (transient receptor potential) mutant of Drosophila melanogaster. This result suggests that La and trp mutation affect the same cellular processes in the photoreceptors.     

Comparative study of visuo-vestibular conditioning in Lymnaea stagnalis

In this review, we compare the current understanding of visuo-vestibular conditioning in Hermissenda crassicornis and Lymnaea stagnalis on the basis of behavioral, electrophysiologic, and morphologic studies. Paired presentation of a photic conditioned stimulus (CS) and an orbital rotation unconditioned stimulus (US) results in conditioned escape behavior in both species. In Hermissenda, changes in excitability of type B photoreceptors and morphologic modifications at the axon terminals follow conditioning. Caudal hair cells, which detect mechanical turbulence, have reciprocal inhibition with type B photoreceptors. In Lymnaea, the interaction between photoreceptors and hair cells is dependent on statocyst location. Furthermore, the organization of the Lymnaea eye is complex, with more than 100 photoreceptors distributed in a uniquely folded retina. Although the optimal conditions to produce long-term memory (memory persistent for >1 week) are almost identical in Hermissenda and Lymnaea, physiologic and morphologic differences suggest that the neuronal mechanisms underlying learning and memory are distinct.     

Different ionic conductances are modulated during the late receptor potential and the prolonged depolarizing afterpotential in Hermissenda type A photoreceptors

Wavelength-dependent, bistable phenomena were found in the receptor potential of Hermissenda crassicornis type A photoreceptors. Short exposure to blue light induced a prolonged depolarizing afterpotential (PDA) following the cessation of the light stimulus. Stronger adaptation to blue light, as caused by prolonged exposure and/or high intensity stimulation, effected a reduction in the early depolarizing transient of the late receptor potential (LRP) as elicited by subsequent stimuli. Vast separation of LRP emergence and PDA emergence could be obtained in photoreceptors in which a strong cancellation of the LRP was accomplished but a PDA still emerged after cessation of the light stimulus. Short exposure to yellow light cancelled the PDA, and stronger adaptation restored the LRP (opposite effect to blue light). The initial depolarizing part of the LRP had earlier been demonstrated to be mediated by the lightdependent increase of an inward conductance. In contrast, in this study the PDA was found to be accompanied by the reduction of an outward conductance, most likely a K⁺ conductance. A bistable photopigment system is thought to control the bistable receptor potential phenomenology by regulating the different membrane conductances during the LRP and the PDA.Abbreviations LRP late receptor potential - PDA prolonged depolarizing afterpotential - PHA prolonged hyperpolarizing afterpotential     

Light-Induced Changes in Photoreceptor Membrane Resistance and Potential in Gecko Retinas : II. Preparations with Active Lateral Interactions

The time-course of light-induced changes in membrane voltage and resistance were measured in single photoreceptors in eyecup preparations of Gekko gekko. A small circular stimulus directed toward the impaled receptor produced membrane hyperpolarization. Application of a steady annular light to the receptor periphery resulted in diminution of the receptor's response to the stimulus. The effects of illumination of the surrounding receptors were isolated by directing a small, steady desensitizing light to the impaled receptor and then applying a peripheral stimulus. Brief stimuli produced a transient decrease in resistance with rapid onset and offset, a time-course similar to that of the response diminution. For some cells a depolarization that coincided with the resistance decrease was seen. During illumination with prolonged stimuli the resistance decrease was followed by a slow increase. After offset resistance rose transiently above the original value and then returned slowly to its original value. The slow resistance changes were not accompanied by changes in membrane voltage. The response diminution, resistance decrease, and depolarization were not observed in retinas treated with aspartate or hypoxia. It is therefore concluded that these effects are mediated by horizontal cells. The diminution is achieved by shunting the receptor potential and may play a role in field adaptation.     

Temporal pattern sensitive and nonsensitive responses in the cat's retinal ganglion cells

In order to characterize temporal pattern sensitivity in the cat ganglion cells, a new analysis technique by semi-Markov models which was developed in the previous papers (Tsukada et al., 1975–1977) was applied to input-output relations of the receptive-field. Three types of statistical spot stimuli positioned in the center region of receptive fields were used. Each type of stimulus has an identical histogram in the inter-stimulus intervals and therefore the same mean and variance, but different correlations between adjacent inter-stimulus intervals (Type 1, positive; Type 2, negative; and Type 3, independent processes). From the output spike trains of cat retinal ganglion cells to each stimulus, mean, variance, and histogram were computed. As the result of investigating these data, we could draw the following conclusion from the resultant output interval histograms. The receptive-field-center responses of cat ganglion cells can be classified into two groups (Types L and N) according to the difference of responsiveness to the three types of statistical spot stimuli. A Type L response has the same histogram in interspike intervals for all three stimuli, and is not sensitive to the temporal pattern, while a Type N response has three different forms depending on each type of stimulus showing high sensitivity to the temporal pattern. These results were also simulated by the Markov chain model and discussed with relation to neural coding and classification of ganglion cell types.     

Contrast gain, signal-to-noise ratio, and linearity in light-adapted blowfly photoreceptors

Response properties of short-type (R1-6) photoreceptors of the blowfly (Calliphora vicina) were investigated with intracellular recordings using repeated sequences of pseudorandomly modulated light contrast stimuli at adapting backgrounds covering 5 log intensity units. The resulting voltage responses were used to determine the effects of adaptational regulation on signal-to-noise ratios (SNR), signal induced noise, contrast gain, linearity and the dead time in phototransduction. In light adaptation the SNR of the photoreceptors improved more than 100-fold due to (a) increased photoreceptor voltage responses to a contrast stimulus and (b) reduction of voltage noise at high intensity backgrounds. In the frequency domain the SNR was attenuated in low frequencies with an increase in the middle and high frequency ranges. A pseudorandom contrast stimulus by itself did not produce any additional noise. The contrast gain of the photoreceptor frequency responses increased with mean illumination and the gain was best fitted with a model consisting of two second order and one double pole of first order. The coherence function (a normalized measure of linearity and SNR) of the frequency responses demonstrated that the photoreceptors responded linearly (from 1 to 150 Hz) to the contrast stimuli even under fairly dim conditions. The theoretically derived and the recorded phase functions were used to calculate phototransduction dead time, which decreased in light adaptation from approximately 5-2.5 ms. This analysis suggests that the ability of fly photoreceptors to maintain linear performance under dynamic stimulation conditions results from the high early gain followed by delayed compressive feed-back mechanisms.     

Electrophysiological Properties of Cells in the Median Ocellus of Limulus

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.     

Maintenance of Motility Bias during Cyanobacterial Phototaxis

Signal transduction in bacteria is complex, ranging across scales from molecular signal detectors and effectors to cellular and community responses to stimuli. The unicellular, photosynthetic cyanobacterium Synechocystis sp. PCC6803 transduces a light stimulus into directional movement known as phototaxis. This response occurs via a biased random walk toward or away from a directional light source, which is sensed by intracellular photoreceptors and mediated by Type IV pili. It is unknown how quickly cells can respond to changes in the presence or directionality of light, or how photoreceptors affect single-cell motility behavior. In this study, we use time-lapse microscopy coupled with quantitative single-cell tracking to investigate the timescale of the cellular response to various light conditions and to characterize the contribution of the photoreceptor TaxD1 (PixJ1) to phototaxis. We first demonstrate that a community of cells exhibits both spatial and population heterogeneity in its phototactic response. We then show that individual cells respond within minutes to changes in light conditions, and that movement directionality is conferred only by the current light directionality, rather than by a long-term memory of previous conditions. Our measurements indicate that motility bias likely results from the polarization of pilus activity, yielding variable levels of movement in different directions. Experiments with a photoreceptor (taxD1) mutant suggest a supplementary role of TaxD1 in enhancing movement directionality, in addition to its previously identified role in promoting positive phototaxis. Motivated by the behavior of the taxD1 mutant, we demonstrate using a reaction-diffusion model that diffusion anisotropy is sufficient to produce the observed changes in the pattern of collective motility. Taken together, our results establish that single-cell tracking can be used to determine the factors that affect motility bias, which can then be coupled with biophysical simulations to connect changes in motility behaviors at the cellular scale with group dynamics.     

Neural Organization of a Molluscan Visual System

Intracellular recording was used to study the response to light of second order visual cells within the optic ganglion of Hermissenda crassicornis. Simultaneous recordings revealed that type B but not type A photoreceptors inhibit the second order cells. Additional details of the neural organization of the visual system were obtained. Possible functional implications of this neural organization are discussed.     

Interactions between hypoxia and hypercapnic acidosis on calcium signaling in carotid body type I cells

The effects of hypercapnic acidosis and hypoxia on intracellular Ca(2+) concentration ([Ca(2+)](i)) were determined with Indo 1 in enzymatically isolated single type I cells from neonatal rat carotid bodies. Type I cells responded to graded hypoxic stimuli with graded [Ca(2+)](i) rises. The percentage of cells responding was also dependent on the severity of the hypoxic stimulus. Raising CO(2) from 5 to 10 or 20% elicited a significant increase in [Ca(2+)](i) in the same cells as those that responded to hypoxia. Thus both stimuli can be sensed by each individual cell. When combinations of hypoxic and acidic stimuli were given simultaneously, the responses were invariably greater than the response to either stimulus given alone. Indeed, in most cases, the response to hypercapnia was slightly potentiated by hypoxia. These data provide the first evidence that the classic synergy between hypoxic and hypercapnic stimuli observed in the intact carotid body may, in part, be an inherent property of the type I cell.     

On the characteristic of nonlinear spatial summation observed in the lateral geniculate cells of cats

The spatial summation characteristic in the receptive fields of cat lateral geniculate cells were investigated. First, the central area of the receptive field was determined using a spot of light. Then the response of the cell were obtained using disc-shaped stimuli of various radii located in the middle point of the receptive field center. When the radius was increased gradually, the response tended to increase, at first, until it reached a peak value and began to decrease thereafter. The radius where the peak response took place was generally less than that of the receptive field center. Furthermore, this radius decreased when the intensity of the stimulus light was increased. These neurophysiological findings could be simulated by a model. The model consists of two parts. The first part receives the input from the photoreceptors. It is of homogeneous structure with shunting inhibition. The second part receives the input from the first part. The structure is characterized by the conventional center-surround type lateral interaction.     

Serial conditional discrimination and temporal bisection in rats selectively lesioned in the dentate gyrus

In positive serial conditional discrimination, animals respond during a target stimulus when it is preceded by a feature stimulus, but they do not respond when the same target stimulus is presented alone. Moreover, the feature and target stimuli are separated from each other by an empty interval. The present work aimed to investigate if two durations (4 or 16 s) of the same feature stimulus (light) could modulate the operant responses of rats to different levers (A and B) during a 5-s target stimulus (tone). In the present study, lever A was associated with the 4-s light, and lever B was associated with the 16-s light. A 5-s empty interval was included between the light and the tone. In the same training procedure, the rats were also presented with the 5-s tone without the preceding light stimuli. In these trials, the responses were not reinforced. We evaluated the hippocampal involvement of these behavioral processes by selectively lesioning the dentate gyrus with colchicine. Once trained, the rats were submitted to a test using probe trials without reinforcement. They were presented with intermediate durations of the feature stimulus (light) to obtain a temporal bisection curve recorded during the exposure to the target stimuli. The rats from both groups learned to respond with high rates during tones preceded by light and with low rates during tones presented alone, which indicated acquisition of the serial conditional discrimination. The rats were able to discriminate between the 4- and 16-s lights by correctly choosing lever A or B. In the test, the temporal bisection curves from both experimental groups showed a bisection point at the arithmetic mean between 4 and 16 s. Such processes were not impaired by the dentate gyrus lesion. Thus, our results showed that different durations of a feature stimulus could result in conditional properties. However, this processing did not appear to depend on the dentate gyrus alone.     

Fine structure of the photogenous areas in the bioluminescent ophiuroidAmphipholis squamata (Echinodermata,Ophiuridea)

 Amphipholis squamata is a small bioluminescent ophiuroid whose arms are the only body part to produce light. The morphology of the arms was described paying particular attention to the spinal ganglia, viz the areas of most intense luminescence. Spinal ganglia consist of five different cell types (A–E) which were studied at different stages of the photogenous reaction. Type D cells have numerous irregularlyshaped vacuoles, widespread Golgi apparatus and well-developed rough endoplasmic reticulum (RER) that show obvious ultrastructural changes after luminescence. Type D cells appear, therefore, to be the best photocyte candidate. Type B and C cells were frequently observed in the nervous system outside spinal ganglia. Type A and E cells have not been described before. Type A cells are ciliated cells and type E cells extend long processes which are intimately associated with type D cells and epidermal ciliated cells. Both type A and type E cells could take part to the stimulation pathway that triggers luminescence. Accepted: 1 September 1996     

A dual synaptic effect on hair cells in Hermissenda

Type A photorecptors can produce an initial hyperpolarizing wave followed by a delayed long-lasting increase in firing which is usually accompanied by a small depolarizing wave. The initial hyperpolarizing wave arises from an increase in conductance while the depolarizing wave was shown to arise from a decrease in conductance. The data presented indicate that both effects produced by the type A photoreceptors in ipsilateral hair cells are synaptic.     

An examination of the effectiveness of inflation and deflation treatments in detecting within-compound learning of a taste aversion

Two conditioned taste aversion experiments with rats assessed the relative effectiveness in providing evidence of within-compound learning of different procedures that involve the initial compound presentation of two stimuli, A and X, with the unconditioned stimulus (i.e., AX+). In Experiment 1, following a single AX+ trial, groups A+ and B+ received an additional conditioning trial (i.e., inflation treatment) with A and B, respectively, whereas group A- received an extinction trial (i.e., deflation treatment) with A. The results showed a reduction in the aversion elicited by the target stimulus, X, in group A- relative to both groups A+ and B+, which did not differ. Experiment 2 further investigated the failure of group A+ to increase the aversion to X relative to control group B+ by pairing A or B with either the same unconditioned stimulus that was previously paired with AX (groups A+ and B+) or with a stronger unconditioned stimulus (groups A* and B*). The results showed increased aversion to X in group A* relative to group B*, but not in group A+ relative to group B+. These results are interpreted as indicative of extinction of the within-compound association during the treatment with A, which could likely impair the detection of within-compound learning following an inflation, but not a deflation treatment.     

Coexpression of spectrally distinct rhodopsins in Aedes aegypti R7 photoreceptors

The retina of the mosquito Aedes aegypti can be divided into four regions based on the non-overlapping expression of a UV sensitive Aaop8 rhodopsin and a long wavelength sensitive Aaop2 type rhodopsin in the R7 photoreceptors. We show here that another rhodopsin, Aaop9, is expressed in all R7 photoreceptors and a subset of R8 photoreceptors. In the dorsal region, Aaop9 is expressed in both the cell body and rhabdomere of R7 and R8 cells. In other retinal regions Aaop9 is expressed only in R7 cells, being localized to the R7 rhabdomere in the central and ventral regions and in both the cell body and rhabdomere within the ventral stripe. Within the dorsal-central transition area ommatidia do not show a strict pairing of R7-R8 cell types. Thus, Aaop9 is coexpressed in the two classes of R7 photoreceptors previously distinguished by the non-overlapping expression of Aaop8 and Aaop2 rhodopsins. Electroretinogram analysis of transgenic Drosophila shows that Aaop9 is a short wavelength rhodopsin with an optimal response to 400-450 nm light. The coexpressed Aaop2 rhodopsin has dual wavelength sensitivity of 500-550 nm and near 350 nm in the UV region. As predicted by the spectral properties of each rhodopsin, Drosophila photoreceptors expressing both Aaop9 and Aaop2 rhodopsins exhibit a uniform sensitivity across the broad 350-550 nm light range. We propose that rhodopsin coexpression is an adaptation within the R7 cells to improve visual function in the low-light environments in which Ae. aegypti is active.     

Correlated receptor and motorneuron changes during retention of associative learning of Hermissenda crassicornis

1. Responses to light of an identified motorneuron (LP1) were recorded simultaneously with those of an identified Hermissenda photoreceptor (the lateral Type B) following three days of training with paired light and rotation. 2. These responses were significantly different when compared to responses of cells from animals trained with unpaired stimuli and from naive animals. 3. The differences of the LP1 responses can be explained as a consequence of the photoreceptor response changes. 4. The same training with paired stimuli has been shown to produce behavioural changes which satisfy criteria for vertebrate associative learning. 5. The observed neural correlates are consistent with previous findings which indicate that membrane changes within the Type B cell bodies play a causal role in associative learning of the nudibranch mollusc, Hermissenda crassicornis.     

Role of stimulus-stimulus pairing in matching-to-sample procedure: cross-species comparison of humans and pigeons

The present experiment examined whether, in a matching-to-sample (MTS) procedure, a relation between two stimuli, a sample and a comparison, could be established as a result of just stimulus–stimulus pairing, even if back up reinforcers were never provided for the conditional relation between the sample and comparison stimuli, but rather only for the comparison stimulus. A procedure called “pseudo matching-to-sample” was used in which, when S1 was presented as a sample stimulus, two comparison stimuli (C1 and C2) were presented, and only responses to C1 were reinforced. Conversely, when S2 was presented, only responses to C3 (and not C4) were reinforced. In other words, organisms experiencing this procedure could discriminate C1 from C2, and C3 from C4, by simple discrimination without regard to the conditional sample stimuli. In order to examine cross-species differences, responding by humans in this procedure was compared to that by pigeons. Although the humans developed a discriminative function for the sample stimuli, that is, the humans’ responding was affected by both the sample stimuli and the reinforcers, responding by the pigeons was affected solely by the reinforcers. These data suggest that, in this procedure, humans (but not pigeons) are able to learn relations among stimuli simply as a result of stimulus–stimulus pairing.     

Immunocytochemical studies on prolactin cells in the adenohypophysis of the golden hamster

Mammotrophs or prolactin (PRL) cells were identified in the adenohypophysis of adult golden hamsters by immunocytochemical techniques with a polyclonal anti-PRL, that was proved to be specific to PRL by the dot immunoblotting test. Postembedding immunostaining was performed on Araldite thin sections by immunoperoxidase and immunogold methods. PRL cells were classified into three types according to the different size of the secretory granules. The Type A cells were usually small and angular or oval in shape, and had secretory granules ranging in diameter from 100-230 nm, and showed poorly developed organelles. The Type B and C cells were larger and round or ovoid in shape, contained larger granules, 230-280 nm and 280-570 nm, respectively, and displayed well developed organelles. Immunoreactive PRL cells in the male pituitaries were far less numerous than in the nonpregnant female glands, and were mostly of the Type A and B, whereas in the female the Type C and B cells predominated. In pregnant females, Type C cells became activated and increased in number, while the other two types decreased in proportion. In lactating females, Type A and B cells significantly increased in number at the expense of the Type C cells; meanwhile, the exocytosis of secretory granules was frequently found in all types of PRL cells. The present findings suggest that Type C and B PRL cells, especially the former, are potent in producing and releasing PRL and highly responsive to various physiological stimuli, while Type A cells are probably relatively inert in synthetic activity.