Research Articles
Immunocytochemical evidence for the presence of histamine and GABA in photoreceptors of the barnacle (Balanus nubilus)
- Joseph C. Callaway, Ann E. Stuart, John S. Edwards
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- 02 June 2009, pp. 289-299
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Biochemical evidence indicates that GABA and histamine may both be synthesized by barnacle photoreceptors (Koike & Tsuda, 1980; Timpe & Stuart, 1984; Callaway & Stuart, 1989b). We used antisera against GABA- and histamine-protein conjugates to determine whether the photoreceptors contain either or both of these antigens. Both antisera labeled all of the photoreceptors in each of the three ocelli. Histamine-like immunoreactivity was found throughout each photoreceptor cell but was most intense at their presynaptic terminals. Histamine-like immunoreactivity was blocked by preincubation of the antibody either with histamine or with a histamine-protein conjugate. GABA-like immunoreactivity was found in all parts of the photoreceptors including the cell body, axon, rhabdomeric dendrites, and presynaptic terminals. GABA-protein conjugates blocked the GABA-like labeling of the photoreceptors, while protein conjugates with histamine, L-glutamate, L-glutamine, β-alanine, and taurine did not. Histamine-like immunoreactivity in the supraesophageal ganglion was confined to the photoreceptor terminals and a second, loose plexus of endings in the main neuropil. GABA-like immunoreactivity, in contrast, was found in approximately twenty-five pairs of neurons of this ganglion. In the cirral nerves, which are expected to contain inhibitory motoneurons, unidentified axons also labeled with the GABA antiserum.
Characterization of guanylate cyclase in squid photoreceptors
- Phyllis R. Robinson, Richard H. Cote
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- 02 June 2009, pp. 1-7
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Light causes a rapid, 1.7-fold increase in cyclic GMP concentration in intact squid retinas (Johnson et al. (1986)). To determine whether light-induced changes in cyclic GMP concentration result from activation of guanylate cyclase, we have studied the regulation of guanylate cyclase activity in squid (Loligo pealei) photoreceptors. The enzyme is membrane-associated and activity is enhanced by the detergents Triton X-100 or digitonin. The enzyme requires divalent cations, Mn2+ being preferred over Mg2+. The dependence of enzyme activity on the MnGTP concentration deviates from simple Michaelis-Menten kinetics. Under conditions where a light-induced binding of GTP to the guanine nucleotide regulatory protein can be observed, no light-induced change in guanylate cyclase could be detected.
Darkness stimulates rapid synthesis and release of melatonin in rat retina
- Dianna A. Redburn, Cheryl K. Mitchell
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- 02 June 2009, pp. 391-403
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The presence of melatonin in retina has been widely reported for over two decades although studies of its functional importance within the retina have only recently been emphasized. We have analyzed the biochemical characteristics of melatonin synthesis and release, focusing on rapid changes in response to light/dark conditions. Our major findings are consistent with the following conclusions: (1) melatonin synthesis is stimulated within minutes after exposure to darkness, and may reflect an increase in N-acetyl transferase activity; (2) melatonin is not stored, but rather it diffuses freely throughout the retina immediately after it is synthesized; and (3) the dark-induced increase in retinal melatonin release is a synthesis-coupled response and does not involve separate secretion mechanisms. The characteristics of melatonin synthesis and release described herein would be consistent with the proposed role of melatonin as a local paracrine effector of dark-adaptive responses in retina.
Octopamine modulates photoreceptor function in the Limulus lateral eye
- George H. Renninger, Robert Schimmel, Claudia A. Farrell
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- 02 June 2009, pp. 83-94
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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.
Changing distribution of GABA-like immunoreactivity in pigeon visual areas during the early posthatching period and effects of retinal removal on tectal GABAergic systems
- Paola Bagnoli, Gigliola Fontanesi, Peter Streit, Luciano Domenici, Roberto Alesci
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- 02 June 2009, pp. 491-508
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The distribution of GABA-like immunoreactivity in the pigeon visual system was studied during the first 9 days after hatching using a mouse monoclonal antibody, mAb 3A12, to glutaraldehyde linked GABA (Matute & Streit, 1986). GABA-like immunoreactivity was seen in cell bodies as well as in neuropil at the level of both the retina and central visual regions at any posthatching age. However, the distribution of putative GABAergic cells and processes varied with age reaching the adult pattern at around 9 days. As a general observation, almost no cell bodies in the retina (except for some perikarya in the ganglion cell layer) were labeled at hatching but densely packed immunostained processes were present in the inner plexiform layer. During the next few days, GABA-immunoreactive amacrine and horizontal cells appeared and the adult distribution of GABA-like immunoreactivity was reached at around 9 days. In the other visual regions examined, the general trend in the variation of GABA-like immunoreactivity included: (1) a progressive decrease in the density of immunostained cell bodies and (2) an almost parallel increase in the concentration of stained neuropil. Since in pigeons the adult organization of visual pathways and the final distribution putative GABAergic systems are reached at around the same age, we suggest the possibility that incoming ganglion cell axons play a role in regulating the distribution of GABA-like immunoreactivity in Visual areas. This hypothesis is supported by the fact that the distribution of GABA-like immunoreactivity in the superficial layers of the optic tectum was altered following ablation of the contralateral retina immediately after hatching.
Review
Centrifugal pathways to the retina: Influence of the optic tectum
- Hiroyuki Uchiyama
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- 02 June 2009, pp. 183-206
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Two types of centrifugal pathways to the retina have been found in the vertebrates, according to the location of the cell bodies and presence or absence of connections with the optic tectum. One type is represented by the isthmo-optic nucleus (ION) of birds and, therefore, termed “ION-type” retinopetal system. The other type is termed “non-ION-type” retinopetal system. The ION-type retinopetal systems have been found in the cyclostomes, teleosts, reptiles, and birds. This review describes the anatomy and physiology of the ION-type retinopetal systems, mainly of birds and teleosts. On the basis of anatomical and physiological evidence cited in this review, the ION-type retinopetal systems can be regarded as the tectofugal pathways to the retina. The function of the ION-type retinopetal systems is discussed in detail, with special emphasis on their relation to the role of the tectum in mediating visuomotor behavior.
Research Articles
Light-regulated proteins in Limulus ventral photoreceptor cells
- Samuel C. Edwards, Anne C. Wishart, Eric M. Wiebe, Barbara-Anne Battelle
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- 02 June 2009, pp. 95-105
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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.
Topography and homogeneity of monkey VI studied through subdurally recorded pattern-evoked potentials
- Gislin Dagnlie, Henk Spekreijse, Bob van Dijk
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- 02 June 2009, pp. 509-525
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Using small checkerboard stimulus fields, we have recorded visually evoked potentials (VEPs) in an alert rhesus monkey from an array of 35 electrodes chronically implanted between dura and arachnoid to study mass neuronal activity in striate and peristriate visual cortex. Although the principal purpose of this work was to study in detail cortical mapping in this particular animal for future intracortical recordings, we report here the usefulness of our approach for the non-invasive study of cortical processing, in particular of cortical magnification and receptive-field properties over the central 6° of the visual field.
The striate and extrastriate components in the pattern onset VEP both have a double negative-going waveform, with N–P–N peak latencies of 75–100–135 ms and 90–115–160 ms, respectively, for small element sizes and moderate contrasts; latencies may be 5 ms shorter for large element sizes and high contrast. We found little activity at electrode locations over visual areas beyond V2. The waveforms and timing permit some careful speculation concerning intracortical processing and VEP generation.
The complex logarithmic form of the retinotopical projection provides a satisfactory model for our data, if a value of 1–1.2° is used for the offset parameter a. Our data suggest that the most abundant receptive-field size in foveal striate cortex has a center diameter of 12′. This size remains constant up to 2° eccentricity, and increases only slowly up to 4°. The smallest receptive-field sizes seem to be independent of eccentricity, throughout the central 4° of Vl, with a value of 4–8′, in agreement with single-cell data reported by Dow et al. (1981) and Van Essen et al. (1984).
Localization of GABA- and GAD-like immunoreactivity in the turtle retina
- Lawrence B. Hurd II, William D. Eldred
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- 02 June 2009, pp. 9-20
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γ-aminobutyric acid (GABA) has been reported to be an important neurotransmitter in the retinas of many species. This immunocytochemical study detailed the localization of antigens resembling GABA and glutamic acid decarboxylase (GAD, an enzyme involved in the synthesis of GABA), in retinal neurons in the turtle, Pseudemys scripta elegans. GABA-like immunoreactivity was present within somata in the inner and outer regions of the inner nuclear layer, within somata in the ganglion cell layer, and in processes in the outer plexiform layer, inner plexiform layer, and ganglion cell axon layer. GAD-like immunoreactivity was found in somata in the inner and outer regions of the inner nuclear layer and in processes in the inner and outer plexiform layers. Cell counts indicated more somata with GABA-like than GAD-like immunoreactivity in the inner nuclear layer. Double-label studies showed that every somata in the inner nuclear layer which had GAD-like immunoreactivity also had GABA-like immunoreactivity, but that many somata had only GABA-like immunoreactivity.
The stratification of immunoreactivity within the inner plexiform layer was analyzed using a scanning densitometer. We described the strata within the inner plexiform layer such that S0 represented the inner nuclear layer/inner plexiform layer border and S100 represented the inner plexiform layer/ganglion cell layer border. Analysis of GAD-like labeling yielded seven distinct strata with peak densities at positions S8, S19, S28, S42, S59, S75, and S93. GABA-like labeling provided five distinct strata with peak densities at positions S17, S28, S67, S84, and S95. The strata with peaks of GABA-like immunoreactivity at S17 and S28 were in statistically identical locations to corresponding strata with GAD-like immunoreactivity. The strata with GABA-like immunoreactivity at S67, S84, and S95 did not have statistically identical peaks of correlated GAD-like immunoreactivity, although there were corresponding strata with GAD-like immunoreactivity nearby. Antiserum directed against GABA failed to produce labeled strata at positions corresponding to the strata with GAD-like immunoreactivity at S8 and S42.
In summary, our results indicated that the antisera we used, which were directed against GABA and GAD, produced significantly different labeling in the inner nuclear layer, inner plexiform layer, and the ganglion cell body and axon layers of the turtle retina. Until the physiological significance of these differences is resolved, studies employing these markers to investigate the function of GABA in the turtle retina should be interpreted with caution.
Research Article
Effects of CNQX, APB, PDA, and kynurenate on horizontal cells of the tiger salamander retina
- Xiong-Li Yang, Samuel M. Wu
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- 02 June 2009, pp. 207-212
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Effects of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-4-phosphonobutyrate (APB), cis-2,3-piperidine dicarboxylic acid (PDA), and kynurenate (KYN) on the depolarizing actions of glutamate and kainate on horizontal cells (HCs) were studied in the larval tiger salamander retina. APB, PDA, and KYN hyperpolarized the HCs, but they failed to block either the actions of glutamate and kainate, or the HC light responses. APB and PDA did not cause membrane polarizations in either rods or cones, suggesting that the HC hyperpolarizations were not mediated by presynaptic actions of these compounds. CNQX, the newly synthesized non-NMDA (N-Methyl-D-Aspartate) receptor antagonist, blocked the HC light responses and the action of kainate, but not that of glutamate. These results suggest that the synaptic receptors in the tiger salamander HCs are probably non-NMDA although extra-synaptic NMDA receptors may exist in these cells.
Research Articles
Comparison of the responses to light and to GABA of cells postsynaptic to barnacle photoreceptors (I-cells)
- Joseph C. Callaway, Ann E. Stuart
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- 02 June 2009, pp. 301-310
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We tested the hypothesis that gamma-aminobutyric acid (GABA) is the transmitter released by barnacle photoreceptors onto postsynaptic cells (I-cells). GABA was applied to I-cells either by superfusion or by ejecting it with pressure from a pipette positioned close to the I-cell's soma. The I-cell's response to GABA was compared with its response to light (i.e. to the photoreceptors' transmitter) by recording intracellularly from its soma. Bath-applied (100 µm to 10 mM) and pressure-applied GABA (10 mM in pipette) hyperpolarizes I-cells by increasing their conductance, as does the photoreceptors' transmitter. The response to pressure-applied GABA consists of two components; both persist when Co2+ or Cd2+ are added to the saline to block synaptic transmission in the preparation, indicating that GABA affects the I-cell directly rather than affecting a presynaptic cell. GABA hyperpolarizes the I-cell when applied to the cell over the soma and ipsilateral arbor or over the contralateral arbor. The I-cells' responses to GABA and to light both depend on extracellular K+ and are affected by changes in intracellular and extracellular Cl−. However, picrotoxin and β-guanidinopropionic acid block the response to pressure-applied GABA but do not block the response to light even at an order of magnitude higher concentration. Thus, GABA is not likely to be the transmitter that causes the hyperpolarizing response of the I-cell. It may be a neuromodulator or the transmitter of an unknown input to the I-cell.
Evolutionary remodeling in a visual system through extensive changes in the synaptic connectivity of homologous neurons
- S. R. Shaw, D. Moore
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- 02 June 2009, pp. 405-410
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The cellular mechanisms by which nervous systems evolve to match evolutionary changes occurring in the rest of the body remain largely unexplored. In a distal visual neuropil of a previously unexamined ancient dipteran family, Stratiomyidae, homologues of all of the periodic neurons known already from more recent Diptera can be recognized, occupying the same locations within the unit structure. This points to extreme developmental stasis for more than 200 million years, conserving both cell identity and position. The arborizations that some neurons make also have remained conservative, but others show marked differences between families in both size and branching patterns. At the electron-microscopical level, extensive differences in synaptic connectivity are found, some sufficient to radically redefine the systems roles of particular neurons. The findings bear out an earlier prediction that changes in the connectivity matrix linking conserved neurons may have been a major factor in implementing evolutionary change in the nervous system.
Pharmacological actions of peptides and indoleamines on turtle retinal ganglion cells
- Alan R. Adolph
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- 02 June 2009, pp. 411-423
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In the turtle retina the peptides met-enkephalin (metENK), somatostatin (SS), neurotensin (NT), and the indoleamine serotonin (5-HT) modulate ganglion cell (GC) activity. The predominant action of the peptides is excitatory, generally enhancing spontaneous firing and light-evoked activity. In contrast, 5-HT usually inhibits these GC activities. MetENK has both direct synaptic input onto GC and indirect action possibly via a GABA inhibitory interneuron. The metENK actions appear mediated via a mu-opiate receptor; morphine and D-ala-metENK-amide (DALA), a stable analog of metENK, are agonists. Naloxone antagonizes the actions of metENK and its agonists. DALA occasionally inhibits GC. This inhibition is antagonized by picrotoxin, while concurrent excitatory action on GC is enhanced. DALA enhances GC response at high spatial frequencies; naloxone attenuates it. The enhancement by DALA suggests a narrowed receptive-field (RF) center, possibly due to changes in a GABA-mediated inhibitory surround. 5-HT inhibitory actions are also mediated via direct and indirect synaptic pathways. 5-methoxy-dimethyl-tryptamine and methoxy-phenyl-piperazine are agonists of 5-HT action. They are both specific 5-HT1 agonists. LSD (lysergic acid diethylamide) and cyproheptadine, which act on 5-HT2 receptors, antagonize 5-HT actions in this retina. Strychnine enhances GC activity, probably by antagonizing glycine-mediated inhibitory inputs. It does not block the inhibitory action of 5-HT, which suggests that the indirect 5-HT inhibition is not mediated via a glycinergic interneurone. 5-HT suppresses directional selectivity (DS) and attenuates high spatial frequencies in some GC. This may be mediated via inhibition of GABAergic amacrines subserving DS and the RF inhibitory surround.
Research Article
Responses to sinusoidal gratings of two types of very nonlinear retinal ganglion cells of cat
- J. B. Troy, G. Einstein, R. P. Schuurmans, J. G. Robson, Ch. Enroth-Cugell
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- 02 June 2009, pp. 213-223
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Perhaps 35% of all of the ganglion cells of the cat do not have classical center-surround organized receptive fields. This paper describes, quantitatively, the responses of two such cell types to stimulation with sinusoidal luminance gratings, whose spatial frequency, mean luminance, contrast, and temporal frequency were varied independently. The patterns were well-focused on the retina of the anesthetized and paralyzed cat. In one type of cell, the maintained discharge was depressed or completely suppressed when a contrast pattern was imaged onto the receptive field (suppressed-by-contrast cell). In the other type of cell, the introduction of a pattern elicited a burst of spikes (impressed-by-contrast cell).
When stimulated with drifting gratings, the cell's mean rate of discharge was reduced (suppressed-by-contrast cell) or elevated (impressed-by-contrast cell) over a limited band of spatial frequencies. There was no significant modulated component of response. The reduction in mean rate of suppressed-by-contrast cells caused by drifting gratings had a monotonic dependence on contrast, a relatively low-pass temporal-frequency characteristic and was greater under photopic than mesopic illuminance. If gratings of spatial frequency, that when drifted evoked a response from these cells, were instead held stationary and contrast-reversed, the mean rate of a suppressed-by-contrast cell was also reduced and that of an impressed-by-contrast cell increased. But, for contrast-reversed gratings, the discharge contained substantial modulation at even harmonic frequencies, the largest being the second harmonic. The amplitude of this second harmonic did not depend on the spatial phase of the grating, and its dependence on spatial frequency, at least for suppressed-by-contrast cells, was similar to that of the reduction in mean rate of discharge. Our results suggest that the receptive fields of suppressed-by-contrast and impressed-by-contrast cells can be modeled with the general form of the nonlinear subunit components of Hochstein and Shapley's (1976) Y cell model.
Research Articles
Calcium/calmodulin-stimulated phosphorylation of photoreceptor proteins in Limulus
- Eric M. Wiebe, Anne C. Wishart, Samuel C. Edwards, Barbara-Anne Battelle
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- 02 June 2009, pp. 107-118
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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.
Quantitative aspects of synaptic ribbon formation in the outer plexiform layer of the developing cat retina
- David H. Rapaport
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- 02 June 2009, pp. 21-32
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The development of synaptic ribbons in rod and cone photoreceptor terminals of the cat retina was studied using quantitative electron microscopy. At the region of the area centralis, synaptic ribbon profiles are initially recognized at PCD (postconception day) 59. Synaptic ribbon density increases rapidly, reaching a peak of 0.55 ribbons/μm3 at PCD 68 (postnatal day 3) and maintains approximately that value for an additional 8 d. Following PCD 76, ribbon density begins to decrease, to 0.37 ribbons/μm3 at PCD 82 and 0.25 ribbons/μm3 at PCD 102. Although ribbon density drops by approximately 50% during this 39-d period, the outer plexiform layer (OPL) volume at the area centralis increases by about 20%. Ribbon density continues to decrease gradually over a protracted period to reach a final adult value of 0.11–0.14 ribbons/μm3. During the period of high ribbon density, rod spherules with two, or even three ribbon profiles, were routinely observed. In contrast, in the adult, spherules with more than one ribbon profile are only rarely encountered. During development, the length of synaptic ribbon profiles increases from a mean of 0.22 μm at PCD 62 to the 0.47 μm mean length found in the adult.
Biochemical and physiological evidence that histamine is the transmitter of barnacle photoreceptors
- Joseph C. Callaway, Ann E. Stuart
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- 02 June 2009, pp. 311-325
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We tested the hypothesis that histamine is the transmitter released by barnacle photoreceptors. Median and lateral ocelli were incubated with 3H-histidine and found to synthesize 3H-histamine, identified by high-voltage electrophoresis. Synthesis could be blocked by the histidine decarboxylase inhibitor (S)-α-fluoromethylhistidine. Histamine was applied to 1-cells either by superfusion or by pressure ejection from a pipette (100 µM or 1 mM histamine) positioned close to the I-cell's soma. When bath-applied at concentrations ranging from 5–100 µM, histamine hyperpolarized the I-cell in a dose-dependent fashion and increased its conductance. At 100 µM, histamine abolished the I-cell's response to light. The response to a pulse of pressure-applied histamine was a hyperpolarization whose amplitude was graded with dose (determined by the duration of the pulse). This response persisted in concentrations of Co2+ and Cd2+ that blocked synaptic transmission from the photoreceptors. Cimetidine, an antagonist of mammalian H2 receptors, markedly decreased the cell's responses both to HA and to light at 100 µM and blocked both responses at 1 mM. Pyrilamine and triprolidine, H1 antagonists, had a complex effect on the I-cell's responses to histamine and to light. Neither H1 nor H2 antagonists, nor histamine itself, affected the voltage or light responses recorded in the presynaptic terminal region, or any phase of calcium-dependent action potentials induced in the terminal in the presence of tetraethylammonium ion. Thus, biochemical, immunocytochemical, and physiological evidence suggests that HA is the transmitter from these photoreceptors to the I-cells. Although gammaaminobutyric acid (GABA) is also present in the photoreceptors, it did not affect the I-cell's responses to light or to histamine when bath-applied at 100 µM. Thus, GABA does not appear to modulate transmission from the photoreceptor to the I-cell.
Expression and cellular localization of substance P/neurokinin A and neurokinin B mRNAs in the rat retina
- Nicholas C. Brecha, Catia Sternini, Karl Anderson, James E. Krause
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- 02 June 2009, pp. 527-535
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The mammalian tachykinin peptides, substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) are encoded by distinct mRNAs derived from separate preprotachykinin (PPT) genes. The SP/NKA-encoding PPT gene generates three mRNAs by alternative RNA processing: α-PPT mRNA, which encodes SP only, and β- and γ-PPT mRNAs, which encode both SP and NKA. The NKB-encoding PPT gene generates mRNAs that produce NKB. The distribution and cellular localization of SP, NKA and NKB mRNAs in the rat retina were studied by RNA blot and in situ hybridization techniques. Blot hybridization analysis of retinal RNA extracts with [32P]-labeled RNA probes complementary to SP/NKA and NKB mRNAs demonstrated single bands of hybridization at 1300 and 900 bases, respectively. Solution hybridization-nuclease protection experiments showed multiple SP/NKA-encoding transcripts with relative levels of ρ-PPT mRNA > β-PPT mRNA ≫ α-PPT mRNA. In situ hybridization histochemistry with [35S]-labeled antisense RNAs demonstrated thatSP/NKA-encoding transcripts are expressed in small-to-medium somata located in the proximal inner nuclear, inner plexiform, and ganglion cell layers, whereas NKB-encoding transcripts are expressed in small-to-medium somata located only in the ganglion cell layer. In this layer, cells containing NKB mRNAs are more numerous than those containing SP/NKA mRNAs. Only background labeling was observed in sections incubated with sense RNA probes, pretreated with RNase A prior to hybridization or incubated in hybridization buffer without the labeled probe. Immunohistochemical studies with a monoclonal antibody directed to the conserved COOH-terminal sequence of the tachykinin peptides revealed tachykinin-like immunoreactive somata with similar size and distribution to those containing SP/NKA- and NKB-encoding transcripts. These results indicate that both SP/NKA and NKB mRNAs are present in the rat retina and that the PPT genes are differentially expressed in specific cell populations. The size and distribution of these cells suggest that they are amacrine and displaced amacrine cells, however, the possibility that tachykinins are present also in ganglion cells in the rat retina cannot be ruled out.
Effects of dark maintenance on retinal biochemistry and function during taurine depletion in the adult rat*
- Susan E. Cocker, Norma Lake
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- 02 June 2009, pp. 33-38
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Light dependence of the effects of taurine depletion on retinal function and biochemistry was examined in albino rats housed either in cyclic lighting or in continuous darkness. Measurements of retinal taurine, DNA, and rhodopsin contents, and electroretinogram amplitudes were made at weekly intervals. Naka-Rushton parameters were estimated for the b wave amplitude-intensity function. No significant effects of lighting regime were observed on retinal taurine levels in untreated rats, or on the time course or extent of taurine depletion in animals treated with guanidinoethyl sulfonate, an antagonist of taurine transport. In both lighting schemes, treatment led to a linear reduction of retinal taurine content which plateaued after 5–6 weeks at 50% of control despite continued treatment. DNA values did not differ among groups, whereas rhodopsin levels doubled in both groups of dark-maintained rats. For treated rats housed in cyclic lighting, the onset of electroretinogram deficits paralleled the loss of retinal taurine in the absence of changes in rhodopsin levels or cell death. Vmax was significantly reduced after 4 weeks of treatment. In contrast, for rats housed in continuous darkness, there were no significant differences in electroretinogram parameters between control and taurine-depleted rats until after 10–14 weeks of treatment. This implies that light exposure accelerates the appearance of functional changes associated with retinal taurine deficiency. The basis of the light dependence and the interpretation of related studies is discussed.
Retinal projections in the ground squirrel (Citellus tridecemlineatus)
- Seema Agarwala, Heywood M. Petry, Jack G. May III
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- 02 June 2009, pp. 537-549
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The retinal projections of the thirteen-lined ground squirrel were determined by tracing anterograde transport of intravitreally injected horseradish peroxidase (HRP) or wheat-germ conjugated horseradish peroxidase (WGA-HRP). Label was seen in the suprachiasmatic nucleus and adjacent anterior hypothalamic area, the accessory optic system (the medial, dorsal, and lateral terminal nuclei), the dorsal and ventral lateral geniculate nuclei, the intergeniculate leaflet, the pretectal nuclei (the anterior, posterior, and olivary pretectal nuclei and the nucleus of optic tract), and the superior colliculus. Most of these structures were labeled bilaterally, with dense contralateral label and sparse ipsilateral label, a pattern typical for animals with laterally placed eyes. However, the suprachiasmatic nucleus and the nucleus of the optic tract received input only from the contralateral eye. In contrast to previous degeneration studies, the sensitive HRP tracers (in conjunction with cytochrome-oxidase reactivity) revealed an elaborate organization within the lateral geniculate nucleus (dorsal LGN, ventral LGN, and intergeniculate leaflet) that is consistent with existing organizational schemes for other mammalian species.