Epileptogenesis alters gene expression pattern in rats subjected to amygdala-dependent emotional learning.

Here we tested a hypothesis that epileptogenesis influences expression pattern of genes in the basolateral amygdala that are critical for fear conditioning. Whole genome molecular profiling of basolateral rat amygdala was performed to compare the transcriptome changes underlying fear learning in epileptogenic and control animals. Our analysis revealed that after fear conditioning procedure 26 genes were regulated differently in the basolateral amygdala of both groups. Thus, our study provides the first evidence that not only the damage to the neuronal pathways but also altered composition or activity level of molecular machinery responsible for formation of emotional memories within surviving pathways can contribute to impairment in emotional learning in epileptogenic animals. Understanding the function of those genes in emotional learning provides an attractive avenue for identification of novel drug targets for treatment of emotional disorders after epileptogenesis-inducing insult.

Endopiriform nucleus connectivities: the implications for epileptogenesis and epilepsy.

Several anterograde and retrograde tracing studies have provided detailed information on the afferent and efferent projections as well as the intrinsic connectivities of the endopiriform nucleus (EN). Here, we summarise EN connectional data and the principles of their organisation and discuss the role they may play in the development and spread of epileptic seizures.

The connections of the endopiriform nucleus with the insular claustrum in the rat and rabbit.

The connections between two parts of the claustrum in the rat and rabbit were studied using the highly fluorescent lipophilic carbocyanine dye (Dil). After the application of Dil crystal into the endopiriform nucleus, labeled fibers in the insular claustrum were observed in its part directly neighboring the insular cortex and capsula externa. Additionally, numerous projections into the piriform, insular and entorhinal cortices were present. The presence of connections between the endopiriform nucleus and insular claustrum suggests its role concerned with the processes taking part in the allocortical regions as well as in the limbic system.

Claustrocingulate connections in the rabbit and rat--a stereological study.

Unbiased stereological methods were used for estimating the numerical density and the total number of claustral neurones projecting to the cingulate cortex in rabbit and rat. In rat the numerical density of neurones projecting to the retrosplenial granular cortex (RSG) differed significantly from those projecting to the retrosplenial agranular (RSA) and cingulate (Cg) cortices while in rabbit the numerical densities of retrogradely labelled neurones in the claustrum following injections into various areas of the cerebral cortex did not differ significantly. The total number of retrogradely labelled neurones in the claustral limbic zones did not differ significantly in both species. The quantitative analysis of claustral zones projecting to a different cingulate cortex area, both in rabbit and rat, reveals that each of these zones is rather homogeneous.

The limbic zone of the rabbit and rat claustrum: a study of the claustrocingulate connections based on the retrograde axonal transport of fluorescent tracers.

The claustrum is a subcortical structure lying under the insular and piriform cortices, whose function is still not clear. Although data exist on connections of the claustrum and the limbic cortex, the topography of the limbic zone in the rabbit and rat claustrum has not been studied extensively. The study was performed on 17 adult Wistar rats and 12 New Zealand rabbits. Two percent water solutions of fluorescent retrograde tracers fast blue and nuclear yellow were injected into the various regions of the limbic cortex. The limbic zone is localized throughout the whole rostrocaudal extent of the claustrum, mainly in its ventromedial portion lying close to the external capsule. Although this zone of the claustrum is localized similarly in both rat and rabbit, some differences between these two species exist. In the rat, neurons projecting to all limbic areas are localized mainly in the anterior and central parts of the claustrum, whereas in the rabbit, the majority of the neurons projecting to the cingulate cortex are present in the anterior and central parts of this structure, while neurons sending axons to the retrosplenial cortex are localized in the central and posterior parts. In both species, double-labeling study showed that neurons projecting to various limbic regions are intermingled and that neurons sending axons into two different limbic regions are seen only occasionally. Our findings give support to the role of the claustrum in integrating information between different areas of the cerebral cortex and the limbic system.

Relationship of calcium-binding protein containing neurons and projection neurons in the rat basolateral amygdala.

Two-laser and two-color approaches were used to observe the colocalization of the calcium-binding proteins, calbindin D28k and parvalbumin, and the retrograde tracer, Fluoro-Gold (FG) in the basolateral amygdala of the rat. The study was performed on five adult rats into which FG was injected to the frontal association cortex. Then, the localization of the retrogradely labeled neurons in the basolateral amygdala was compared with the localization of the neurons labeled by calcium-binding proteins. The present study showed that most of the retrogradely labeled neurons in the posterior part of the basolateral amygdala are also calbindin-positive. Even though a lot of parvalbumin-positive endings were present at the surface of the retrogradely labeled cells, we did not observe the colocalization of the parvalbumin and projective neurons.

Parvalbumin immunoreactivity changes in the thalamic reticular nucleus during the maturation of the rat's brain.

The thalamic reticular nucleus (Rt) is a thin lamina of cells, through which thalamocortical and corticothalamic fibers pass. It is interposed between the thalamic nuclei and the internal capsule and it is composed of GABA-ergic cells with synapses that receive impulses from both kinds of fibers. Rt takes part in the negative feed-back system of controlling the information transfer from the thalamus to the cerebral cortex and it is focused in the sleep-waking cycle. The pattern of parvalbumin reactivity during maturation of Rt becomes the main aim of our study. The study was performed on 36 rats on various postnatal days (P0, P1, P2, P4, P5, P7, P10, P14, P17, P21, P30 and P90). The animals were anesthetized, transcardially perfused, cut on cryostat into 30-microns-thick frontal sections, stained immunocytochemically using standard ABC method and a mouse monoclonal antibody against parvalbumin. A small amount of round and oval, parvalbumin immunopositive cells was detected at stage PO, predominantly in the intermediate part of Rt, whereas the cells in ventral and lateral part at the same time were only slightly immunopositive. At P10 the cells in the intermediate part became more fusiform or oval because of the appearance of dendrites. At P14 we were able to observe separate, punctuated structures interpreted as the axonal endings. There were plenty of them at the time of full maturation of the intermediate portion of reticular nucleus (stage P21). At this time, the dorsal and ventral parts had their first synapses, too, but their maturation ended a week later. At P30 multipolar neurons, with round and fusiform somata were distributed relatively homogeneously throughout Rt. We compared the stage with the parvalbumin reactivity of the adult rat and found no difference in the morphological pattern of PV neurons.

The projection of the amygdaloid nuclei to various areas of the limbic cortex in the rat.

The connections of the amygdaloid body with the areas of the limbic cortex in the rat were studied by means of the method on the axonal retrograde transport of the fluorescent tracer FluoroGold. The tracer was injected into the anterior and posterior limbic cortices (cingulate gyrus, granular and agranular retrosplenial area, respectively. The localization of the corresponding amygdalar projection zones was investigated and the semiquantitative analysis of the connections was conducted. The projection zones in the rat amygdaloid body are organized topographically. Administration of the fluorescent tracer to the anterior and posterior part of the limbic cortex in the rat (cingulate gyrus and both retrosplenial areas, respectively) reveals labeling of cells only for injections to the former one. The labeled cells were present only in the major components of the basolateral amygdaloid complex. The main source of this projection was anterior part of basolateral nucleus (BLA). Some labeled neurons were found in the lateral nucleus and few in the ventral part of basolateral nucleus. No labeled cells were found within the basomedial nucleus.