Investigation of multi-site micro recordings of subthalamic nucleus neurons using machine learning MER with DBS in Parkinson`s – A simulation study

Multineural spikes were acquired with a multisite electrode placed in the hippocampus pyramidal cell layer of non-primate anesthetized snitch animals. If the impedance of each electrode-site is relatively low and the distance amongst electrode sites is appropriately miniatured, a spike generated by a neuron is parallelly recorded at multielectrode sites with different amplitudes. The covariance between the spike of the at each electrode-point and a template was computed as a damping-factor due to the volume conduction of the spike from the neuron to electrode-site. Computed damping factors were vectorized and analyzed by simple but elegant hierarchical-clustering using a multidimensional statistical-test. Since a cluster of damping vectors was shown to correspond to an antidromically identified neuron, spikes of distinct neurons are classified by suggesting to the scatterings of damping vectors. Errors in damping vector computing due to partially overlapping spikes were minimized by successively subtracting preceding spikes from raw data. Clustering errors due to complex-spike-bursts (i.e., spikes with variable-amplitudes) were prevented by detecting such bursts and using only the first spike of a burst for clustering.

Recording from an Identified Neuron Efficiently Reveals Hazard for Brain Function in Risk Assessment

Modern societies use a continuously growing number of chemicals. Because these are released into the environment and are taken up by humans, rigorous (but practicable) risk assessment must precede the approval of new substances for commerce. A number of tests is applicable, but it has been very difficult to efficiently assay the effect of chemicals on communication and information processing in vivo in the adult vertebrate brain. Here, we suggest a straightforward way to rapidly and accurately detect effects of chemical exposure on action potential generation, synaptic transmission, central information processing, and even processing in sensory systems in vivo by recording from a single neuron. The approach is possible in an identified neuron in the hindbrain of fish that integrates various sources of information and whose properties are ideal for rapid analysis of the various effects chemicals can have on the nervous system. The analysis uses fish but, as we discuss here, key neuronal functions are conserved and differences can only be due to differences in metabolism or passage into the brain, factors that can easily be determined. Speed and efficiency of the method, therefore, make it suitable to provide information in risk assessment, as we illustrate here with the effects of bisphenols on adult brain function.

Bisphenols exert detrimental effects on neuronal signaling in mature vertebrate brains

Bisphenols are important plasticizers currently in use and are released at rates of hundreds of tons each year into the biosphere1–3. However, for any bisphenol it is completely unknown if and how it affects the intact adult brain4–6, whose powerful homeostatic mechanisms could potentially compensate any effects bisphenols might have on isolated neurons. Here we analyzed the effects of one month of exposition to BPA or BPS on an identified neuron in the vertebrate brain, using intracellular in vivo recordings in the uniquely suited Mauthner neuron in goldfish. Our findings demonstrate an alarming and uncompensated in vivo impact of both BPA and BPS—at environmentally relevant concentrations—on essential communication functions of neurons in mature vertebrate brains and call for the rapid development of alternative plasticizers. The speed and resolution of the assay we present here could thereby be instrumental to accelerate the early testing phase of next-generation plasticizers.

Specific labeling of synaptic schwann cells reveals unique cellular and molecular features

Perisynaptic Schwann cells (PSCs) are specialized, non-myelinating, synaptic glia of the neuromuscular junction (NMJ), that participate in synapse development, function, maintenance, and repair. The study of PSCs has relied on an anatomy-based approach, as the identities of cell-specific PSC molecular markers have remained elusive. This limited approach has precluded our ability to isolate and genetically manipulate PSCs in a cell specific manner. We have identified neuron-glia antigen 2 (NG2) as a unique molecular marker of S100β+ PSCs in skeletal muscle. NG2 is expressed in Schwann cells already associated with the NMJ, indicating that it is a marker of differentiated PSCs. Using a newly generated transgenic mouse in which PSCs are specifically labeled, we show that PSCs have a unique molecular signature that includes genes known to play critical roles in PSCs and synapses. These findings will serve as a springboard for revealing drivers of PSC differentiation and function.

Acute regulation of habituation learning via posttranslational palmitoylation

Habituation is an adaptive learning process that enables animals to adjust innate behaviors to changes in the environment. Despite its well documented implications for a wide diversity of behaviors, the molecular and cellular basis of habituation learning is not well understood. Using whole genome sequencing of zebrafish mutants isolated in an unbiased genetic screen, we identified the palmitoyltransferase Hip14 as a critical regulator of habituation learning. We demonstrate that Hip14 regulates depression of sensory inputs onto an identified neuron and provide compelling evidence that Hip14 palmitoylates the Shaker-like channel subunit Kv1.1, thereby regulating Kv1.1 subcellular localization. Furthermore, we show that loss of either Kv1.1 or Hip14 leads to habituation deficits, and that Hip14 is dispensable in development and instead acts acutely to promote habituation. Combined, our results uncover a previously unappreciated role for acute post-translational palmitoylation at defined circuit components to regulate learning.

When complex neuronal structures may not matter

Much work has explored animal-to-animal variability and compensation in ion channel expression. Yet, little is known regarding the physiological consequences of morphological variability. We quantify animal-to-animal variability in cable lengths (CV = 0.4) and branching patterns in the Gastric Mill (GM) neuron, an identified neuron type with highly-conserved physiological properties in the crustacean stomatogastric ganglion (STG) of Cancer borealis. We examined passive GM electrotonic structure by measuring the amplitudes and apparent reversal potentials (Erevs) of inhibitory responses evoked with focal glutamate photo-uncaging in the presence of TTX. Apparent Erevs were relatively invariant across sites (mean CV ± SD = 0.04 ± 0.01; 7–20 sites in each of 10 neurons), which ranged between 100–800 µm from the somatic recording site. Thus, GM neurons are remarkably electrotonically compact (estimated λ > 1.5 mm). Electrotonically compact structures, in consort with graded transmission, provide an elegant solution to observed morphological variability in the STG.

Morphology of Invertebrate Neurons and Synapses

Despite their often small numbers, the neurons in invertebrate nervous systems can nevertheless constitute many classes, and the nervous systems of little studied or entirely new species still offer significant opportunities for discovery. Circuit analyses and connectomic data are of particular significance, as are the relationships of these to behavior, and the organization of simple larval brains. Functional analyses of synaptic circuits still require knowledge of the neurotransmitter and neurotransmitter receptor for each identified neuron. Synapse complexity ranges widely; undifferentiated pathways in basal species may have unpolarized synapses with presynaptic sites opposite each other, and specialized pathways may have polyadic synapses.

Inhibitory modulation of optogenetically identified neuron subtypes in the rostral solitary nucleus

Inhibition is presumed to play an important role in gustatory processing in the rostral nucleus of the solitary tract (rNST). One source of inhibition, GABA, is abundant within the nucleus and comes both from local, intrasolitary sources and from outside the nucleus. In addition to the receptor-mediated effects of GABA on rNST neurons, the hyperpolarization-sensitive currents, Ih and IA, have the potential to further modulate afferent signals. To elucidate the effects of GABAergic modulation on solitary tract (ST)-evoked responses in phenotypically defined rNST neurons and to define the presence of IA and Ih in the same cells, we combined in vitro recording and optogenetics in a transgenic mouse model. This mouse expresses channelrhodopsin 2 (ChR2) in GAD65-expressing GABAergic neurons throughout the rNST. GABA positive (GABA+) neurons differed from GABA negative (GABA−) neurons in their response to membrane depolarization and ST stimulation. GABA+ neurons had lower thresholds to direct membrane depolarization compared with GABA− neurons, but GABA− neurons responded more faithfully to ST stimulation. Both IA and Ih were present in subsets of GABA+ and GABA− neurons. Interestingly, GABA+ neurons with Ih were more responsive to afferent stimulation than inhibitory neurons devoid of these currents, whereas GABA− neurons with IA were more subject to inhibitory modulation. These results suggest that the voltage-gated channels underlying IA and Ih play an important role in modulating rNST output through a circuit of feedforward inhibition.