Toxoplasma gondii actively inhibits neuronal function in chronically infected mice.

Upon infection with the obligate intracellular parasite Toxoplasma gondii, fast replicating tachyzoites infect a broad spectrum of host cells including neurons. Under the pressure of the immune response, tachyzoites convert into slow-replicating bradyzoites, which persist as cysts in neurons. Currently, it is unclear whether T. gondii alters the functional activity of neurons, which may contribute to altered behaviour of T. gondii-infected mice and men. In the present study we demonstrate that upon oral infection with T. gondii cysts, chronically infected BALB/c mice lost over time their natural fear against cat urine which was paralleled by the persistence of the parasite in brain regions affecting behaviour and odor perception. Detailed immunohistochemistry showed that in infected neurons not only parasitic cysts but also the host cell cytoplasm and some axons stained positive for Toxoplasma antigen suggesting that parasitic proteins might directly interfere with neuronal function. In fact, in vitro live cell calcium (Ca(2+)) imaging studies revealed that tachyzoites actively manipulated Ca(2+) signalling upon glutamate stimulation leading either to hyper- or hypo-responsive neurons. Experiments with the endoplasmatic reticulum Ca(2+) uptake inhibitor thapsigargin indicate that tachyzoites deplete Ca(2+) stores in the endoplasmatic reticulum. Furthermore in vivo studies revealed that the activity-dependent uptake of the potassium analogue thallium was reduced in cyst harbouring neurons indicating their functional impairment. The percentage of non-functional neurons increased over time In conclusion, both bradyzoites and tachyzoites functionally silence infected neurons, which may significantly contribute to the altered behaviour of the host.

Non-sensory cortical and subcortical connections of the primary auditory cortex in Mongolian gerbils: bottom-up and top-down processing of neuronal information via field AI.

In the present study, we will provide further anatomical evidence that the primary auditory cortex (field AI) is not only involved in sensory processing of its own modality, but also in complex bottom-up and top-down processing of multimodal information. We have recently shown that AI in the Mongolian gerbil (Meriones unguiculatus) has substantial connections with non-auditory sensory and multisensory brain structures [Budinger, E., Heil, P., Hess, A., Scheich, H., 2006. Multisensory processing via early cortical stages: Connections of the primary auditory cortical field with other sensory systems. Neuroscience 143, 1065-1083]. Here we will report about the direct connections of AI with non-sensory cortical areas and subcortical structures. We approached this issue by means of the axonal transport of the sensitive bidirectional neuronal tracers fluorescein-labelled (FD) and tetramethylrhodamine-labelled dextran (TMRD), which were simultaneously injected into different frequency regions of the gerbil's AI. Of the total number of retrogradely labelled cell bodies found in non-sensory brain areas, which identify cells of origin of direct projections to AI, approximately 24% were in cortical areas and 76% in subcortical structures. Of the cell bodies in the cortical areas, about 4.4% were located in the orbital, 11.1% in the infralimbic medial prefrontal (areas DPC, IL), 18.2% in the cingulate (3.2% in CG1, 2.9% in CG2, 12.1% in CG3), 9.5% in the frontal association (area Fr2), 12.0% in the insular (areas AI, DI), 10.8% in the retrosplenial, and 34.0% in the perirhinal cortex. The cortical regions with retrogradely labelled cells, as well as the entorhinal cortex, also contained anterogradely labelled axons and their terminations, which means that they are also target areas of direct projections from AI. The laminar pattern of corticocortical connections indicates that AI receives primarily cortical feedback-type inputs and projects in a feedforward manner to its target areas. The high number of double-labelled somata, the non-topographic distribution of single FD- and TMRD-labelled somata, and the overlapping spatial distribution of FD- and TMRD-labelled axonal elements suggest rather non-tonotopic connections between AI and the multimodal cortices. Of the labelled cell bodies in the subcortical structures, about 38.8% were located in the ipsilateral basal forebrain (10.6% in the lateral amygdala LA, 11.5% in the globus pallidus GP, 3.7% in the ventral pallidum VPa, 13.0% in the nucleus basalis NB), 13.1% in the ipsi- and contralateral diencephalon (6.4% in the posterior paraventricular thalamic nuclei, 6.7% in the hypothalamic area), and 48.1% in the midbrain (20.0% in the ipsilateral substantia nigra, 9.8% in the ipsi- and contralateral ventral tegmental area, 5.0% in the ipsi- and contralateral locus coeruleus, 13.3% the ipsi- and contralateral dorsal raphe nuclei). Thus, the majority of subcortical inputs to AI was related to different neurotransmitter systems. Anterograde labelling was only found in some ipsilateral basal forebrain structures, namely, the LA, basolateral amygdala, GP, VPa, and NB. As for the cortex, the proportion and spatial distribution of single FD-, TMRD-, and double-labelled neuronal elements suggests rather non-tonotopic connections between AI and the neuromodulatory subcortical structures.

Manganese-enhanced MRI reveals structural and functional changes in the cortex of Bassoon mutant mice.

Manganese-enhanced magnetic resonance imaging (ME-MRI) was used to analyze the brain architecture in mice lacking the functional presynaptic active zone protein Bassoon. Anatomical characterization revealed a significant increase in the total brain volume in Bassoon mutants as compared with wild-type mice, which is mainly caused by changes in cortex and hippocampus volume. The measured enlargement in cortical volume coincides with an altered Mn2+ distribution within cortical layers as visualized by T1-weighted magnetic resonance imaging. Two days after manganese application, the cortex of Bassoon mutant mice appeared more laminated in ME-MRI, with an enhanced accumulation of manganese in deep, central, and superficial cortical cell layers. Whereas morphologically the cortical lamination is not affected by the absence of a functional Bassoon, an altered basal activation pattern was found in the cortex of the mutant mice both by metabolic labeling with [14C]-2-deoxyglucose and histochemical detection of the potassium analogue thallium uptake. Consequently, the results indicate that the absence of the functional presynaptic protein Bassoon causes disturbance in the formation of normal basal cortical activation patterns and thereby in the functional cortical architecture. Furthermore, this study shows that ME-MRI can become a valuable tool for a structural characterization of genetically modified mice.