SK3 trafficking in hippocampal cells: the role of different molecular domains.

The regulative steps that control trafficking of ion channels are fundamental determinants of their qualitative and quantitative expression on the cell membrane. In this work the trafficking of the small conductance calcium-activated potassium channel, SK3 was studied in neurons in order to identify relevant molecular domains involved in this process. Hippocampal cell cultures were transfected with fusion proteins of green fluorescent protein (GFP) and different SK3 subunit truncations. The differential distribution of the mutants was analyzed by confocal microscopy and compared to the localization of the control fusion protein with full length SK3. The transport of chimeric proteins was quantified from fluorescence images by developing a morphometric analytical method. We found that the full length SK3 was distributed in cell body, axon and dendrites, whereas the deleted forms GFPDelta578-736 (deletion of the entire C-terminal domain), GFPDeltaCaMBD (deletion of the calmodulin-binding site) and GFPDeltaN (deletion of the N-terminal domain) were not transported into cell processes but accumulated in the cell body. The GFPDelta640-736 (deletion of the distal C-terminal domain) showed a distribution similar to control. The quantification and statistical analysis confirmed the differences in distribution across the three groups. In conclusion, the current work provides evidence for a fundamental role of the N-terminal domain and the calmodulin binding domain in SK3 trafficking in neurons.

Antagonism at serotonin 5-HT(2A) receptors modulates functional activity of frontohippocampal circuit.

RATIONALE: Several second-generation antipsychotics are characterised by a significant antagonistic effect at serotonin 5-HT(2A) receptors (5-HT(2A)R), a feature that has been associated with lower incidence of extra-pyramidal symptoms and a putative amelioration of positive and negative symptoms experienced by schizophrenic patients. However, the neurofunctional substrate of 5-HT(2A) antagonism and its exact contribution to the complex pharmacological profile of these drugs remain to be elucidated. OBJECTIVES: Here, we used pharmacological magnetic resonance imaging to map the modulatory effects of the selective 5-HT(2A)R antagonist Ml00907 on the spatiotemporal patterns of brain activity elicited by acute phencyclidine (PCP) challenge in the rat. PCP is a non-competitive NMDA receptor antagonist that induces dysregulation of corticolimbic glutamatergic neurotransmission and produces cognitive impairment and psychotic-like symptoms reminiscent of those observed in schizophrenia. RESULTS: Pre-administration of M100907 produced focal and region-dependent attenuation of PCP-induced response in frontoseptohippocampal areas. As early studies highlighted a permissive role of 5-HT(2A)R on frontal dopamine release, the role of post-synaptic dopamine D(1) receptors on PCP-induced response was examined by using the potent antagonist SCH23390. Interestingly, SCH23390 did not affect PCP's response in any of the regions examined. This finding rules out a significant contribution of dopamine in the functional changes mapped and, indirectly, the inhibitory effect of M100907, in favour of a glutamatergic origin. CONCLUSIONS: Our data expand recent evidence suggesting a key role of 5-HT(2A)R in modulating glutamate-mediated cognitive performance in the prefrontal cortex and highlight the whole frontoseptohippocampal circuit as a key functional substrate of 5-HT(2A)R antagonism in normal and disease states.