A comparison of possible markers for chandelier cartridges in rat medial prefrontal cortex and hippocampus.

Chandelier neurons and their characteristic arrays of axonal terminals, known as cartridges, have been implicated in a variety of psychiatric and neurological disorders including schizophrenia and epilepsy. As a result, these neurons have been extensively examined in the brains of several species using a range of markers. However, these markers have not been systematically compared in a single species for their robustness in labelling chandelier cell cartridges. We have therefore examined several markers, reported to label chandelier arrays in primates, for their capacity to mark these structures in rat medial prefrontal cortex and hippocampus. These studies revealed that cartridge-like structures were labelled by parvalbumin and GAT-1 immunohistochemistry in both medial prefrontal cortex and hippocampus of the rat brain. Additionally, GAD65 immunohistochemistry labelled array-like structures preferentially in the dentate gyrus. In contrast, PSA-NCAM, calbindin and GAD67 immunohistochemistry did not reveal any array-like structures in either region of rat brain. These observations indicate that the various immunological markers previously used to visualise chandelier cell cartridges in primates are not equally efficient in labelling these structures in the rat brain, and that GAT-1 immunohistochemistry is the most robust means of visualising chandelier cell cartridges in the regions examined. These are important considerations for quantitative studies in animal models of neurological disorders where chandelier neurons are implicated.

Quantification of morphological differences in boutons from different afferent populations to the nucleus accumbens.

The nucleus accumbens (Acb) receives convergent glutamatergic inputs from the prefrontal cortex (PFC), central thalamus, basolateral amygdala and the ventral subiculum of the hippocampus. The principal neurons of the nucleus accumbens are modulated by specific sets of convergent afferent inputs, the local circuit neurons also receive a substantial number of glutamatergic inputs, but the full complement of these has yet to be established. The aim of these studies was to define characteristics of the different glutamatergic afferent inputs to the nucleus accumbens that would aid their identification. To enable the characterisation of the glutamatergic inputs to nucleus accumbens neurons we first labelled the four main glutamatergic sources of afferent input to the accumbens with the anterograde tracer biotinylated dextran amine (BDA). Using an unbiased systematic sampling method, the morphological characteristics of their synaptic boutons were measured and assessed at the electron microscopic level. From the criteria assessed, a comparison of the four afferent sources was made, characteristics such as bouton size and vesicle density had significantly different population means, however, the only characteristic that allowed discrimination between the four major glutamatergic afferent to the nucleus accumbens was that of vesicle size. The vesicles in boutons from amygdala were larger than the subiculum which, in turn, were larger than the prefrontal cortex, the thalamus were the smallest in size. The methods used also allow a comparison of the relative frequency of different sized postsynaptic structures targeted, the prefrontal cortex almost exclusively targeted spines whereas the thalamus and the subiculum, in addition to spines, targeted proximal and distal dendrites.

Basolateral amygdala efferents to the ventral subiculum preferentially innervate pyramidal cell dendritic spines.

The basolateral amygdala and the ventral subiculum of the hippocampal formation are two of the major limbic-related regions within the brain, both of which project heavily to the nucleus accumbens. The nucleus accumbens is regarded as the limbic-motor interface, in view of these limbic afferent and its somatomotor and autonomic efferent connections. These afferent inputs have been suggested to converge monosynaptically on cells within the accumbens and are hypothesised to play a role in functions such as affective motivational behaviour. Convergence between inputs from the basolateral amygdala and the hippocampus at the level of the accumbens can be demonstrated with electrophysiological recording methods, but these do not conclusively preclude polysynaptic mechanisms. In fact there is a robust reciprocal projection between the basolateral amygdala and the hippocampus, synaptic details of which have not been fully investigated. We examined the synaptic input from the basolateral amygdala to the projection neurons of the subiculum, the spiny pyramidal neurons. We labelled the afferents from basolateral amygdala with a small injection of biotinylated dextran amine, and revealed the anterogradely labeled fibers within the subiculum. The labeled basolateral amygdala fibers were studied with electron microscopy to identify their postsynaptic target structures. With this technique we have demonstrated anatomically that the basolateral amygdala preferentially innervates spiny subiculum neurons, presumed pyramidal projection neurons, although some dendrites and possibly local circuit neurons may be targeted.