Darrell A. Henze

Interneuron Diversity series: Circuit complexity and axon wiring economy of cortical interneurons

The performance of the brain is constrained by wiring length and maintenance costs. The apparently inverse relationship between number of neurons in the various interneuron classes and the spatial extent of their axon trees suggests a mathematically definable organization, reminiscent of small-world or scale-free networks observed in other complex systems. The wiring-economy-based classification of cortical inhibitory interneurons is supported by the distinct physiological patterns of class members in the intact brain. The complex wiring of diverse interneuron classes could represent an economic solution for supporting global synchrony and oscillations at multiple timescales with minimum axon length.

Integration and Segregation of Activity in Entorhinal-Hippocampal Subregions by Neocortical Slow Oscillations

Brain systems communicate by means of neuronal oscillations at multiple temporal and spatial scales. In anesthetized rats, we find that neocortical slow oscillation engages neurons in prefrontal, somatosensory, entorhinal, and subicular cortices into synchronous transitions between UP and DOWN states, with a corresponding bimodal distribution of their membrane potential. The membrane potential of hippocampal granule cells and CA3 and CA1 pyramidal cells lacked bimodality, yet it was influenced by the slow oscillation in a region-specific manner. Furthermore, in both anesthetized and naturally sleeping rats, the cortical UP states resulted in increased activity of dentate and most CA1 neurons, as well as the highest probability of ripple events. Yet, the CA3-CA1 network could self-organize into gamma bursts and occasional ripples during the DOWN state. Thus, neo/paleocortical and hippocampal networks periodically reset, self-organize, and temporally coordinate their cell assemblies via the slow oscillation.

Hilar mossy cells: functional identification and activity in vivo.

Network oscillations are proposed to provide the framework for the ongoing neural computations of the brain. Thus, an important aspect of understanding the functional roles of various cell classes in the brain is to understand the relationship of cellular activity to the ongoing oscillations. While many studies have characterized the firing properties of cells in the hippocampal network including granule cells, pyramidal cells and interneurons, information about the activity of dentate mossy cells in the intact brain is scant. Here we review the currently available information and describe biophysical properties and network-related firing patterns of mossy cells in vivo. These new observations will assist in the extracellular identification of this unique cell type and help elucidate their functional role in behaving animals.

Imidazopyridine CB2 agonists: Optimization of CB2/CB1 selectivity and implications for in vivo analgesic efficacy

A new series of imidazopyridine CB2 agonists is described. Structural optimization improved CB2/CB1 selectivity in this series and conferred physical properties that facilitated high in vivo exposure, both centrally and peripherally. Administration of a highly selective CB2 agonist in a rat model of analgesia was ineffective despite substantial CNS exposure, while administration of a moderately selective CB2/CB1 agonist exhibited significant analgesic effects.

Decahydroquinoline amides as highly selective CB2 agonists: Role of selectivity on in vivo efficacy in a rodent model of analgesia

A novel series of decahydroquinoline CB2 agonists is described. Optimization of the amide substituent led to improvements in CB2/CB1 selectivity as well as physical properties. Two key compounds were examined in the rat CFA model of acute inflammatory pain. A moderately selective CB2 agonist was active in this model. A CB2 agonist lacking functional CB1 activity was inactive in this model despite high in vivo exposure both peripherally and centrally.

High concentration electrophysiology-based fragment screen: discovery of novel acid-sensing ion channel 3 (ASIC3) inhibitors.

The Merck Fragment Library was screened versus acid-sensing ion channel 3 (ASIC3), a novel target for the treatment of pain. Fragment hits were optimized using two strategies, and potency was improved from 0.7 mM to 3 μM with retention of good ligand efficiency and incorporation of reasonable physical properties, off-target profile, and rat pharmacokinetics.

Benzimidazole CB2 agonists: design, synthesis and SAR.

A new series of CB2-selective agonists containing a benzimidazole core is reported. Design, synthesis, SAR and pharmacokinetic data for selected compounds are described.

Recent Advances Toward Pain Therapeutics

Publisher Summary This chapter focuses on recent advances toward pain therapeutics. Two of the most promising novel targets actively pursued for chronic pain are the voltage-gated sodium (Nav) channels—in particular Nav 1.3, 1.7, and 1.8—and nerve growth factor (NGF/tropomyosin-related kinase receptor A (TrkA). Both of these targets are predominantly localized in the periphery. The achievement of good target selectivity with small molecules is also a common challenge of both pathways. A summary of recent drug development for these two pathways, highlighting recent developments and progress made toward finding truly selective inhibitors, is provided in the chapter. In contrast to sodium channels and NGF/TrkA, fatty acid amide hydrolase (FAAH) is believed to play a significant role in both the periphery and the central nervous system. The chapter covers some recently disclosed novel noncovalent inhibitors and clinical data that raise questions about the validity of the target. The treatment of chronic pain is potentially on the cusp of entering a new era with a number of novel mechanistic approaches in clinical development. Many of these novel mechanisms present unique medicinal chemistry challenges in developing safe and effective medicines.

Single cell contributions to network activity in the hippocampus

Abstract The information processing capabilities of neuronal populations are a function of the connectivity and single cell properties of the neurons that comprise the network. An important area of investigation is how information is encoded and processed by neurons in the network. There is a growing body of evidence demonstrating how the specific biophysical properties of individual neurons contribute to network processing as a whole. We illustrate through several examples in the hippocampal formation how cellular biophysics can directly effect the timing of neuronal spiking in the context of on-going network oscillations and how network patterns can influence the intrinsic properties of single cells. We discuss the consequences of the cooperation and competition between single cell and network properties.

High-Throughput Screen of GluK1 Receptor Identifies Selective Inhibitors with a Variety of Kinetic Profiles Using Fluorescence and Electrophysiology Assays

GluK1, a kainate subtype of ionotropic glutamate receptors, exhibits an expression pattern in the CNS consistent with involvement in pain processing and migraine. Antagonists of GluK1 have been shown to reduce pain signaling in the spinal cord and trigeminal nerve, and are predicted to provide pain and migraine relief. We developed an ultra-high-throughput small-molecule screen to identify antagonists of GluK1. Using the calcium indicator dye fluo-4, a multimillion-member small-molecule library was screened in 1536-well plate format on the FLIPR (Fluorescent Imaging Plate Reader) Tetra against cells expressing a calcium-permeable GluK1. Following confirmation in the primary assay and subsequent counter-screen against the endogenous Par-1 receptor, 6100 compounds were selected for dose titration to assess potency and selectivity. Final triage of 1000 compounds demonstrating dose-dependent inhibition with IC50 values of less than 12 µM was performed in an automated whole-cell patch clamp electrophysiology assay. Although a weak correlation between electrophysiologically active and calcium-imaging active compounds was observed, the identification of electrophysiologically active compounds with a range of kinetic profiles revealed a broad spectrum of mechanisms of action.