Location of release sites and calcium-activated chloride channels relative to calcium channels at the photoreceptor ribbon synapse.

Vesicle release from photoreceptor ribbon synapses is regulated by L-type Ca(2+) channels, which are in turn regulated by Cl(-) moving through calcium-activated chloride [Cl(Ca)] channels. We assessed the proximity of Ca(2+) channels to release sites and Cl(Ca) channels in synaptic terminals of salamander photoreceptors by comparing fast (BAPTA) and slow (EGTA) intracellular Ca(2+) buffers. BAPTA did not fully block synaptic release, indicating some release sites are <100 nm from Ca(2+) channels. Comparing Cl(Ca) currents with predicted Ca(2+) diffusion profiles suggested that Cl(Ca) and Ca(2+) channels average a few hundred nanometers apart, but the inability of BAPTA to block Cl(Ca) currents completely suggested some channels are much closer together. Diffuse immunolabeling of terminals with an antibody to the putative Cl(Ca) channel TMEM16A supports the idea that Cl(Ca) channels are dispersed throughout the presynaptic terminal, in contrast with clustering of Ca(2+) channels near ribbons. Cl(Ca) currents evoked by intracellular calcium ion concentration ([Ca(2+)](i)) elevation through flash photolysis of DM-nitrophen exhibited EC(50) values of 556 and 377 nM with Hill slopes of 1.8 and 2.4 in rods and cones, respectively. These relationships were used to estimate average submembrane [Ca(2+)](i) in photoreceptor terminals. Consistent with control of exocytosis by [Ca(2+)] nanodomains near Ca(2+) channels, average submembrane [Ca(2+)](i) remained below the vesicle release threshold (∼ 400 nM) over much of the physiological voltage range for cones. Positioning Ca(2+) channels near release sites may improve fidelity in converting voltage changes to synaptic release. A diffuse distribution of Cl(Ca) channels may allow Ca(2+) influx at one site to influence relatively distant Ca(2+) channels.

Synaptology of the inner plexiform layer in the anuran retina.

The inner plexiform layer of the retina is a synaptic layer mostly devoid of perikarya. It contains the processes of three major neuron types: the bipolar cells, which carry information from the photoreceptors, the ganglion cells, which are the output elements of the retina, and the amacrine cells, which are able to influence the communication between the former two. Since amacrine cells are the most diverse retinal neurons, they are in a position to carve out and delineate the neural circuits of the inner retina. The aim of this review is to offer a summary of findings related to the general synaptology of the inner retina in frogs and also to provide some insight into the synaptic organization of neurochemically identified amacrine cells. The main conclusions of this paper are as follows: (i) Most contacts are formed between amacrine cells. (2) Direct bipolar to ganglion cell synapses exist, but are rare in the anuran retina. (3) All neurochemically identified amacrine cell types receive inputs from bipolar cells, but not all of them form reciprocal contacts with bipolar cell axon terminals. (4) A major inhibitory transmitter, gamma-aminobutyric acid, is involved in more than 50% of the synapses. Since contacts between inhibitory elements were often observed, disinhibitory circuits must also play a role in retinal information processing. (5) Reciprocal relationship between dopaminergic and gamma-aminobutyric acid-containing cells have been confirmed. Similar situation was observed in case of serotoninergic and gamma-aminobutyric acid-positive elements. No contacts were verified between serotoninergic and dopaminergic elements. (6) Both monoamine- and neuropeptide-containing amacrine cells establish direct contacts with ganglion cell dendrites, providing a morphological basis for neuromodulatory influence on the output elements of the retina. Unfortunately, only a handful of studies have been carried out to identify the synaptic connections between neurochemically identified cells in the anuran retina. Double-label studies at the electron microscope level to reveal the synaptic relationship of cell populations containing two different transmitters/modulators are extremely rare. Further insight into retinal synaptic circuitries could be gained with a combination of electrophysiology and morphology at the electron microscopic level. These studies must also involve identification of the transmitter receptors on identified cell types. Only after this step can the function of different synaptic circuitries be better approximated.

Comparative study of the neuronal plasticity along the neuraxis of the vibrissal sensory system of adult rat following unilateral infraorbital nerve damage and subsequent regeneration.

The aim of the present study was to examine the physiological consequences of a unilateral infraorbital nerve lesion and its regeneration at different levels of the somatosensory neuraxis. In animals whose right infraorbital nerve had been crushed, a large unresponsive area was found in the main brainstem trigeminal nucleus (Pr5). Responses evoked by ipsilateral vibrissal deflection in the middle of Pr5 reappeared only on days 22-35 after the nerve had been transected, whereas recovery from the nerve crush took only 7-9 days. However, no sign of short-term neuronal plasticity was observed in Pr5 after peripheral nerve injury. An enlargement of the receptive fields in two-thirds of the units and a lengthening in the delay of the evoked responses were observed as long-term plastic changes in Pr5 neurons after peripheral-nerve regeneration. In the ventral posteromedial nucleus of the thalamus (VPM) of partly denervated animals, however, only minutes or hours after the nerve crush, certain units were found to respond in some cases not only to the vibrissae, but also to mechanical stimulation of the face over the eye (two units), the nose (one unit), and the midline (one unit). Apart from the experiments involving incomplete denervation, the vibrissal representation areas of the VPM were unresponsive to stimulation of both the vibrissae and other parts of the face until nerve regeneration had occurred. In the somatosensory cortex, an infraorbital nerve crush immediately resulted in a large cortical area being unresponsive to vibrissal deflection. It was noteworthy, however, that shortly after the nerve crush, this large unresponsive whisker representation cortical area was invaded from the rostromedial direction by responses evoked by stimulation of the forepaw digits. In spite of the reappearance of vibrissa-evoked responses 7-10 days after the nerve crush, an expanded digital representation could still be observed 3 weeks after the nerve crush, resulting in an overlapping area of digital and vibrissal representations. The withdrawal of the expanded representation of forepaw digits was completed by 60 days after the nerve crush. The results obtained in Pr5, the VPM, and the cortex strongly suggest that the higher the station in the neuraxis, the greater the degree of plasticity after infraorbital nerve injury.