In this study we have characterized the membrane properties and morphology of interneurons which lie between the caudal pole of the trigeminal motor nucleus and the rostral border of the facial motor nucleus. Previous studies suggest that many of these interneurons may participate in the genesis of rhythmical jaw movements. Saggital brainstem slices were taken from rats aged 5-8 days. Interneurons lying caudal to the trigeminal motor nucleus were visualized using near-infrared differential interference contrast (DIC) microscopy, and were recorded from using patch pipettes filled with a K-gluconate- and biocytin-based solution. The 127 neurons recorded could be categorized into three subtypes on the basis of their responses to injection of depolarizing current pulses, namely tonic firing (type I), burst firing (type II) and spike-adaptive (type III) neurons. Type I interneurons had a higher input resistance and a lower rheobase than type II neurons. All three neuron subtypes showed 'sag' of the voltage response to injection of large-amplitude hyperpolarizing current pulses, and, in addition, also showed rectification of the voltage response to injection of depolarizing current pulses, with type II neurons showing significantly greater rectification than type I neurons. The axonal arborizations were reconstructed for 44 of 63 neurons labelled with tracer. Neurons of each subtype were found to issue axon collaterals terminating in the brainstem nuclei, including the parvocellular reticular nucleus (PCRt), the trigeminal motor nucleus (Vmot), the supratrigeminal nucleus or the trigeminal mesencephalic nucleus. Twenty-five of the 43 neurons issued collaterals which terminated in the Vmot and the other brainstem nuclei. When viewed under 100x magnification, the collaterals of some interneurons were seen to give off varicosities and end-terminations which passed close to the somata of unidentified neurons in the trigeminal motor nucleus and in the area close to the interneuron soma itself. This suggests that the interneurons may make synaptic contacts both on motoneurons and also on nearby interneurons. These results provide data on the membrane properties of trigeminal interneurons and evidence for their synaptic connections both with nearby interneurons and also with motoneurons. Thus, the interneurons examined could play roles in the shaping, and possibly also in the generation, of rhythmical signals to trigeminal motoneurons.

g Growth-associated protein 43 (GAP43) is known to regulate axon growth, but whether it also plays a role in synaptogenesis remains unclear. Here, we found that GAP43 regulates the aggregation of gephyrin, a pivotal protein for clustering postsynaptic GABA A receptors (GABA A Rs), in developing cortical neurons. Pharmacological blockade of either protein kinase C (PKC) or neuronal activity increased both GAP43-gephyrin association and gephyrin misfolding-induced aggregation, suggesting the importance of PKC-dependent regulation of GABAergic synapses. Furthermore, we found that PKC phosphorylation-resistant GAP43 S41A , but not PKC phosphorylation-mimicking GAP43 S41D , interacted with cytosolic gephyrin to trigger gephyrin misfolding and its sequestration into aggresomes. In contrast, GAP43 S41D , but not GAP43 S41A , inhibited the physiological aggregation/clustering of gephyrin, reduced surface GABA A Rs under physiological conditions, and attenuated gephyrin misfolding under transient oxygen-glucose deprivation (tOGD) that mimics pathological neonatal hypoxia. Calcineurin-mediated GAP43 dephosphorylation that accompanied tOGD also led to GAP43-gephyrin association and gephyrin misfolding. Thus, PKC-dependent phosphorylation of GAP43 plays a critical role in regulating postsynaptic gephyrin aggregation in developing GABAergic synapses. Proper development of inhibitory GABAergic synapses is critical for establishing an excitatory/inhibitory balance in the neural network (1, 2). The impairment of postsynaptic GABA A receptor (GABA A R) activity is a major cause of neuronal hyperactivity, affecting cognitive development and psychosocial behaviors (3, 4). Postsynaptic surface insertion and clustering of GABA A Rs determine the efficacy of GABAergic synapses (4, 5). Gephyrin, a microtubule-associated protein, is a key scaffolding protein that requires the GABA A R ␥2 subunit for clustering GABA A Rs at the postsynaptic membrane (6, 7). The lack of neuronal gephyrin reduces postsynaptic GABA A R clustering, thereby impairing inhibitory synaptic transmission (8, 9).In central neurons, gephyrin monomers oligomerize to form a hexagonal lattice, also called gephyrin clusters, underneath the cell surface membrane to anchor postsynaptic GABA A Rs (10). However, numerous studies have shown that gephyrin is an aggregation-prone protein that forms large clumps when expressed in nonneural cells or cell-free systems (11, 12). Instead, gephyrin in neurons forms small aggregates/clusters in both the cytosol and submembrane domain for receptor clustering, suggesting a neuronal machinery that regulates gephyrin clustering. To date, a postsynaptic protein, collybistin, a GDP-GTP exchanging factor, is the only gephyrin-interacting protein that can effectively disperse gephyrin clumps into oligomeric clusters in HEK293T cells (13). Gephyrin scaffolding in neurons depends on the dynamic rearrangement of microtubules and actin microfilaments at postsynaptic sites (14,15). Whether cytoskeleton-associated proteins are involved in regulat...

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Background Inflammation is a potential risk factor of mental disturbance. FKBP5 that encodes FK506-binding protein 51 (FKBP51), a negative cochaperone of glucocorticoid receptor (GR), is a stress-inducible gene and has been linked to psychiatric disorders. Yet, the role of FKBP51 in the inflammatory stress-associated mental disturbance remained unclear. Methods Fkbp5-deficient (Fkbp5-KO) mice were used to study inflammatory stress by a single intraperitoneal injection of lipopolysaccharide (LPS). The anxiety-like behaviors, neuroimaging, immunofluorescence staining, immunohistochemistry, protein and mRNA expression analysis of inflammation- and neurotransmission-related mediators were evaluated. A dexamethasone drinking model was also applied to examine the effect of Fkbp5-KO in glucocorticoid-induced stress. Results LPS administration induced FKBP51 elevation in the liver and hippocampus accompanied with transient sickness. Notably, Fkbp5-KO but not wild-type (WT) mice showed anxiety-like behaviors 7 days after LPS injection (LPS-D7). LPS challenge rapidly increased peripheral and central immune responses and hippocampal microglial activation followed by a delayed GR upregulation on LPS-D7, and these effects were attenuated in Fkbp5-KO mice. Whole-brain [18F]-FEPPA neuroimaging, which target translocator protein (TSPO) to indicate neuroinflammation, showed that Fkbp5-KO reduced LPS-induced neuroinflammation in various brain regions including hippocampus. Interestingly, LPS elevated glutamic acid decarboxylase 65 (GAD65), the membrane-associated GABA-synthesizing enzyme, in the hippocampus of WT but not Fkbp5-KO mice on LPS-D7. This FKBP51-dependent GAD65 upregulation was observed in the ventral hippocampal CA1 accompanied by the reduction of c-Fos-indicated neuronal activity, whereas both GAD65 and neuronal activity were reduced in dorsal CA1 in a FKBP51-independent manner. GC-induced anxiety was also examined, which was attenuated in Fkbp5-KO and hippocampal GAD65 expression was unaffected. Conclusions These results suggest that FKBP51/FKBP5 is involved in the systemic inflammation-induced neuroinflammation and hippocampal GR activation, which may contribute to the enhancement of GAD65 expression for GABA synthesis in the ventral hippocampus, thereby facilitating resilience to inflammation-induced anxiety.