The balance between excitatory and inhibitory synaptic transmission within and across neurons in active networks is crucial for cortical function and may allow for rapid transitions between stable network states. GABAergic interneurons mediate the majority of inhibitory transmission in the cortex, and therefore contribute to the global balance of activity in neuronal networks. Disruption in the network balance due to impaired inhibition has been implicated in several neuropsychiatric diseases (Marin 2012). Both schizophrenia and autism are two highly heritable cognitive disorders with complex genetic aetiologies but overlapping behavioural phenotypes that share common imbalances in neuronal network activity (Gao & Penzes 2015). An increasing body of evidence suggests that functional abnormalities in a particular group of cortical GABAergic interneurons expressing the calcium-binding protein parvalbumin (PV) are involved in the pathology of these disorders (Marin 2012). As deficits in this neuronal population have been linked to these disorders it could be useful to target them and increase their activity. A conserved feature in PV cells is their unusually low input resistance compared to other neuronal populations. This feature is regulated by the expression of leak K+ channels, believed to be mediated in part by TASK and TREK subfamily two-pore K+ channels (Goldberg et al. 2011). The selective blockade of specific leak K+ channels could therefore be applied to increase the activity of PV cells. In this thesis, specific TASK-1/3 and TREK-1 channel blockers were applied in cortical mouse slices in an attempt to increase the output of PV cells. The blockade of either channel did not successfully increase the amplitude of PV cell-evoked inhibitory postsynaptic currents (IPSCs) onto principal cells. However, while the blockade of TASK-1/3 channels failed to depolarise the membrane or alter the input resistance, the blockade of TREK-1 channels resulted in a small but significant depolarisation of the membrane potential in PV cells. Interestingly, TREK-1 channel blockade also increased action potential firing of PV cells in response to given current stimuli, suggesting that TREK-1 could be a useful target for PV cell modulation. These results demonstrate for the first time the functional effects of using specific two-pore K+ channel blockers in PV cells. Furthermore, these data provide electrophysiological evidence against the functional expression of TASK-1/3 in PV cells. It could therefore be interesting to further characterise the precise subtypes of leak K+ channels responsible for their low resistivity. This would help to classify the key contributors of the background K+ conductances present in PV cells in addition to finding suitable targets to increase their activity.

The subiculum is a structure that forms a bridge between the hippocampus and the entorhinal cortex (EC) in the brain, and plays a major role in the memory consolidation process. It consists of different types of pyramidal neurons. Based on their firing behavior, these excitatory neurons are classified into strong burst firing (SBF), weak burst firing (WBF) and regular firing (RF) neurons. In the first part of the work, morphological differences in the different neuronal subtypes was explored by biocytin staining after classifying the neurons based on the differences in electrophysiological properties. Detailed morphological properties of these three neuronal subtypes were analyzed using Neurolucida neuron reconstruction method. Unlike the differences in their electrophysiological properties, no difference was found in the morphometric properties of these neuronal subtypes. In the second part of the thesis, experimental results on spike- timing- dependent plasticity (STDP) at the proximal excitatory inputs on the subicular pyramidal neurons of the juvenile (P15-P19) rat are described. The STDP was studied in the WBF and RF neurons. Causal pairing of a single EPSP with a single back propagating action potential (bAP) at a time interval of 10 ms failed to induce plasticity. However, increasing the number of bAPs in such EPSP-bAP pair to three at 50 Hz (bAP burst) induced LTD in both, the RF, as well as the WBF neurons. Increasing the frequency of action potentials to 150 Hz in the bAP burst during causal pairing also induced LTD in both the neuronal subtypes. However, all other STDP related experiments were performed only with the bAP bursts consisting of 3 bAPs evoked at 50 Hz. Amplitude of the causal pairing induced LTD decreased with increasing time interval between EPSP and the bAP burst. Reversing the order of the EPSP and the bAP burst in the pair induced LTP only with a short time interval of 10 ms. This finding is in contrast to most of the reports on excitatory synapses, wherein the pre-before post (causal) pairing induced LTP and vice-versa. The results of causal and anti-causal pairing were used to plot the STDP curve for the WBF neurons. In the STDP curve observed in these synapses, LTD was observed upto a causal time interval of 30 ms, while LTP was limited to 10 ms time interval. Hence, the STDP curve was biased towards LTD. These results reaffirm the earlier observations that the relative timing of the pre- and postsynaptic activities can lead to multiple types of STDP curves. Next, the mechanism of non-Hebbian LTD was studied in both, the RF and WBF neurons. The involvement of calcium in the postsynaptic neuron in plasticity induction was studied by chelating intracellular calcium with BAPTA. The results indicate that the LTD induction in WBF neurons required postsynaptic calcium, while LTD induction in the RF neurons was independent of postsynaptic calcium. Paired pulse ratio (PPR) experiments suggested the involvement of a presynaptic mechanism in the induction of LTD in the RF neurons, and not in the WBF neurons since the PPR was unaffected by the induction protocol only in the WBF neurons. LTD induction in the WBF neurons required activity of the NMDA receptors since LTD was not observed in the presence of the NMDA receptor blocker in the WBF neurons, while it was unaffected in the RF neurons. However, the RF neurons required the activity of L-type calcium channels for plasticity induction, since LTD was affected in the presence of the L-type calcium channel blockers, although the WBF neurons did not require the L-type calcium channel activity for plasticity induction. Hence, in addition to a non-Hebbian STDP curve, a novel mechanism of LTD induction has been reported, where L-type calcium channels are involved in a synaptic plasticity that is expressed via change in the release probability. The findings on the STDP in subicular pyramidal neurons may have strong implications in the memory consolidation process owing to the central role of the subiculum and LTD in it.

Neuroligine (NL) sind vorwiegend postsynaptisch lokalisierte transmembrane Adhäsionsmoleküle, die in Wechselwirkung mit dem präsynaptisch lokalisierten Protein Neurexin eine wichtige Rolle in der Reifung und Funktion von Synapsen spielen. Es existieren verschiedene NL-Isoproteine (NL-1 – NL-4), die sich in ihrer Assoziation zu exzitatorischen und inhibitorischen Synapsen unterscheiden. Die funktionelle und klinische Relevanz der Neuroligine belegen beispielhaft Mutationen des Isotyps NL 4, welche mit neuropsychiatrischen Erkrankungen wie Autismus-Spektrum-Störungen assoziiert vorkommen. Anhand eines durch Ausschalten des human-orthologen NL-4-Gens generierten Mausmodells (NL 4 Knockout, NL 4 KO) konnte in vorhergehenden Studien die Bedeutung einer immunhistochemisch beobachteten Lokalisation von NL 4 an glycinergen Synapsen der Retina für die inhibitorische synaptische Übertragung nachgewiesen werden. Im Unterschied dazu konnte kein Zusammenhang zwischen einer in Schicht IV des Barrel-Kortex nachweisbaren Lokalisation von NL-4 mit inhibitorischen Synapsen hergestellt werden. Deshalb, und aufgrund der in Schicht IV dominierenden exzitatorischen Verschaltung von thalamischen Projektionen und den kolumnenassoziierten Rückverschaltungen aus dem Neokortex, lässt sich eine Interaktion von NL-4 mit exzitatorischen Synapsen in diesem Areal vermuten. Im Rahmen der vorliegenden Arbeit wurde anhand der NL-4-KO-Modellmaus der Frage nachgegangen, inwiefern NL-4 die exzitatorische synaptische Übertragung im Barrel-Kortex beeinflusst. Dafür wurden mit Hilfe der Patch-Clamp-Technik abgeleitete AMPA-Rezeptor-vermittelte exzitatorische postsynaptische Ströme (EPSC) von bedornten Sternzellen, Sternpyramiden- und Pyramidenzellen der Schicht IV ausgewertet und zwischen NL-4-Wildtyp- (NL 4-WT) und NL 4 KO-Neuronen verglichen. Dabei zeigten NL 4-KO-Neurone signifikant veränderte Parameter der EPSC-Kinetik. Die Abfallszeit war in NL 4 KO-Neuronen signifikant länger, das maximale Gefälle und die maximale Steigung signifikant flacher gegenüber NL-4-WT-Kontrollen. Diese Veränderungen sprechen für eine funktionelle Relevanz von NL-4 für die AMPA-Rezeptor-vermittelte synaptische Übertragung auf exzitatorische Neurone in Schicht IV des Barrel-Kortex. Das Muster der in NL-4-KO-Neuronen veränderten EPSC-Kinetik weist dabei auf eine Modulation der biophysikalischen AMPA-Rezeptoreigenschaften hin und könnte mit Veränderungen der synaptisch exprimierten AMPA-Rezeptor-TARP-Subtypen in Zusammenhang stehen, die über Proteine der postsynaptischen Dichte (wie PSD-95 und S SCAM) mit Neuroliginen interagieren.