A selective role for neuronal activity regulated pentraxin in the processing of sensory-specific incentive value

Neuronal activity regulated pentraxin (Narp) is a secreted neuronal product which clusters AMPA receptors and regulates excitatory synaptogenesis. Although Narp is selectively enriched in brain, its role in behavior is not known. As Narp is expressed prominently in limbic regions, we examined whether Narp deletion affects performance on tasks used to assess motivational consequences of food-rewarded learning. Narp knock-out (KO) mice were unimpaired in learning simple pavlovian discriminations, instrumental lever pressing, and in acquisition of at least two aspects of pavlovian incentive learning, conditioned reinforcement and pavlovian-instrumental transfer. In contrast, Narp deletion resulted in a substantial deficit in the ability to use specific outcome expectancies to modulate instrumental performance in a devaluation task. In this task, mice were trained to respond on two levers for two different rewards. After training, mice were prefed with one of the two rewards, devaluing it. Responding on both levers was then assessed in extinction. Whereas control mice showed a significant preference in responding on the lever associated with the nondevalued reward, Narp KO mice responded equally on both levers, failing to suppress responding on the lever associated with the devalued reward. Both groups consumed more of the nondevalued reward in a subsequent choice test, indicating Narp KO mice could distinguish between the rewards themselves. These data suggest Narp has a selective role in processing sensory-specific information necessary for appropriate devaluation performance, but not in general motivational effects of reward-predictive cues on performance

Preserved acute pain and impaired neuropathic pain in mice lacking protein interacting with C Kinase 1

Protein interacting with C Kinase 1 (PICK1), a PDZ domain-containing scaffolding protein, interacts with multiple different proteins in the mammalian nervous system and is believed to play important roles in diverse physiological and pathological conditions. In this study, we report that PICK1 is expressed in neurons of the dorsal root ganglion (DRG) and spinal cord dorsal horn, two major pain-related regions. PICK1 was present in approximately 29.7% of DRG neurons, most of which were small-less than 750 μm2 in cross-sectional area. Some of these PICK1-positive cells co-labeled with isolectin B4 or calcitonin-gene-related peptide. In the dorsal horn, PICK1 immunoreactivity was concentrated in the superficial dorsal horn, where it was prominent in the postsynaptic density, axons, and dendrites. Targeted disruption of PICK1 gene did not affect basal paw withdrawal responses to acute noxious thermal and mechanical stimuli or locomotor reflex activity, but it completely blocked the induction of peripheral nerve injury-induced mechanical and thermal pain hypersensitivities. PICK1 appears to be required for peripheral nerve injury-induced neuropathic pain development and to be a potential biochemical target for treating this disorder

Phosphorylation of AMPA Receptors Is Required for Sensory Deprivation-Induced Homeostatic Synaptic Plasticity

Sensory experience, and the lack thereof, can alter the function of excitatory synapses in the primary sensory cortices. Recent evidence suggests that changes in sensory experience can regulate the synaptic level of Ca2+-permeable AMPA receptors (CP-AMPARs). However, the molecular mechanisms underlying such a process have not been determined. We found that binocular visual deprivation, which is a well-established in vivo model to produce multiplicative synaptic scaling in visual cortex of juvenile rodents, is accompanied by an increase in the phosphorylation of AMPAR GluR1 (or GluA1) subunit at the serine 845 (S845) site and the appearance of CP-AMPARs at synapses. To address the role of GluR1-S845 in visual deprivation-induced homeostatic synaptic plasticity, we used mice lacking key phosphorylation sites on the GluR1 subunit. We found that mice specifically lacking the GluR1-S845 site (GluR1-S845A mutants), which is a substrate of cAMP-dependent kinase (PKA), show abnormal basal excitatory synaptic transmission and lack visual deprivation-induced homeostatic synaptic plasticity. We also found evidence that increasing GluR1-S845 phosphorylation alone is not sufficient to produce normal multiplicative synaptic scaling. Our study provides concrete evidence that a GluR1 dependent mechanism, especially S845 phosphorylation, is a necessary pre-requisite step for in vivo homeostatic synaptic plasticity

Selective injection system into hippocampus CA1 via monitored theta oscillation.

Methods of cell biology and electrophysiology using dissociated primary cultured neurons allow in vitro study of molecular functions; however, analysis of intact neuronal circuitry is often preferable. To investigate exogenous genes, viral vectors are most commonly injected using a pipette that is inserted from the top of the cortex. Although there are few reports that describe the success rate of injection in detail, it is sometimes difficult to locate the pipette tip accurately within the CA1 pyramidal cell layer because the pyramidal layer is only 0.1 mm thick. In the present study, we have developed a system to inject viral vectors accurately into the mouse hippocampal CA1 pyramidal cell layer using a stereotaxic injection system with simultaneous electrophysiological monitoring of theta oscillation. The pipette tip was positioned reliably based on integrated values of the theta oscillation in the hippocampal CA1 pyramidal cell layer. This approach allows accurate injection of solutions and provides an efficient method of gene transfer using viral vectors into the hippocampus, which can be a useful tool for studies involving the molecular mechanisms of neuronal functions

Differential expression of isoforms of PSD-95 binding protein (GKAP/SAPAP1) during rat brain development 1The nucleotide sequences reported in this paper have been submitted to the GenBank with accession numbers AB003594, AB005146.1

AbstractPSD-95/SAP90, which binds to the C-terminus of NMDA receptor and Shaker-type potassium channel, is one of the major postsynaptic density proteins. Recently, novel classes of proteins interacting with the guanylate kinase domain of PSD-95 have been identified, guanylate kinase-associated protein (GKAP) and SAP90/PSD-95-associated proteins (SAPAPs). Here we report the isolation of new isoforms of PSD-95 binding protein (GKAP/SAPAP1) using the yeast two-hybrid system. The isolated protein directly interacts with the guanylate kinase domain of PSD-95. Northern blot analyses revealed that the expression of these isoforms containing distinct N-terminal sequences is differentially regulated during brain development. The present findings suggest that each isoform of the PSD-95 binding protein is differentially expressed in a development-dependent manner and may be involved in the complex formation of PSD-95 and channel/receptors at the postsynaptic density

Disordered testosterone transport in mice lacking the ganglioside GM2/GD2 synthase gene

Genetic disruption of glycosyltransferases has provided clear information on the roles of their reaction products in the body. Our group has studied the function of glycosphingolipids by genetic engineering of glycosyltransferases in cell culture and in mice, which has demonstrated both expected and unexpected results. Among these findings, aspermatogenesis in ganglioside GM2/GD2 synthase knockout mice was one of the most surprising and intriguing results. There were no sperms in testis, and multinuclear giant cells were detected instead of spermatids. Although serum levels of testosterone in the male mice were extremely low, testosterone accumulated in the interstitial tissues, including Leydig cells, and seemed not to be transferred into the seminiferous tubules or vascular cavity from Leydig cells. This was considered to be the cause of aspermatogenesis and low serum levels of testosterone. Patients with a mutant GM2/GD2 synthase gene (SPG26) showed similar clinical signs, not only in terms of the neurological aspects, but also in the male reproductive system. The mechanisms for testosterone transport by gangliosides are discussed here based on our own results and reports from other laboratories

Representative EEG data.

<div><p><b>A</b>. Raw EEG data collected from a recording electrode (top) and filtered EEG after selecting for theta oscillations (4-8 Hz) (middle). An isolated theta oscillation in a 10 sec and 20 sec bin was analyzed using integrated values (0.5 sec and 10 sec durations) at a depth of 0.95mm from the cortex. </p> <p><b>B</b>. Sequential recording of EEG at different depths. The same three types of data are presented as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083129#pone-0083129-g002" target="_blank">Figure 2A</a> with changes at different depths indicated at the bottom. Twenty seconds of recording data at each depth were collected, analyzed, and integrated over 0.5 sec and 10 sec.</p></div

Representative traces showing changes in the integrated value of the theta oscillation at different depths.

<div><p><b>A</b>. A theta oscillation integrated over 0.5 sec indicated the highest value presumably in the pyramidal layer of the hippocampus. After the first peak, the value declined towards the bottom of the stratum radiatum and then increased again.</p> <p><b>B</b>. Superimposition of the integrated values of the theta oscillation. The changes in the integrated values across seven independent experiments were superimposed. All of the data presented a similar pattern of peaks, and these peaks overlapped at a point that was assumed to be the CA1 pyramidal layer.</p></div

Electrophysiological analysis of lentivirus-infected neurons.

<p>EGFP-expressing lentivirus vectors were stereotaxically injected into the CA1 pyramidal neurons, and patch clamp analyses were carried out on infected and non-infected neurons. <b>A</b>. <b>B</b>. Analysis of the mEPSCs showed comparable frequency, amplitude, and decay time compared to the non-infected neurons. Non-infected cells: EGFP(-) n=7, infected cells: EGFP(+) n=8, <b>C</b>. Pairing-induced LTP was not significantly different between the two types of neurons. EGFP(-) n=6, EGFP(+) n=7. <b>D</b>. The injection of ZsGreen lentivirus vector expressing GluA1 into hippocampus CA1 had partially rescued LTP in GluA1 knockout mice. (*<i>P<0</i>.<i>05</i>, **<i>P<0.01</i>) <b>E</b>. Statistical analysis of LTP in wild (EGFP (-)), rescued, and GluA1 knockout mice in 10 min and 30 min after LTP induction. LTP was partially rescued by GluA1-expressing lentivirus infection in GluA1 knockout mouse. (*<i>P<0</i>.<i>05</i>, **<i>P<0.01</i>).</p