5' tRNA halves are highly expressed in the primate hippocampus and might sequence-specifically regulate gene expression.

Fragments of mature tRNAs have long been considered as mere degradation products without physiological function. However, recent reports show that tRNA-derived small RNAs (tsRNAs) play prominent roles in diverse cellular processes across a wide spectrum of species. Contrasting the situation in other small RNA pathways the mechanisms behind these effects appear more diverse, more complex, and are generally less well understood. In addition, surprisingly little is known about the expression profiles of tsRNAs across different tissues and species. Here, we provide an initial overview of tsRNA expression in different species and tissues, revealing very high levels of 5' tRNA halves (5' tRHs) particularly in the primate hippocampus. We further modulated the regulation capacity of selected 5' tRHs in human cells by transfecting synthetic tsRNA mimics ("overexpression") or antisense-RNAs ("inhibition") and identified differentially expressed transcripts based on RNA-seq. We then used a novel k-mer mapping approach to dissect the underlying targeting rules, suggesting that 5' tRHs silence genes in a sequence-specific manner, while the most efficient target sites align to the mid-region of the 5' tRH and are located within the CDS or 3' UTR of the target. This amends previous observations that tsRNAs guide Argonaute proteins to silence their targets via a miRNA-like 5' seed match and suggests a yet unknown mechanism of regulation. Finally, our data suggest that some 5' tRHs that are also able to sequence-specifically stabilize mRNAs as up-regulated mRNAs are also significantly enriched for 5' tRH target sites.

Inhibitory interneuron classes express complementary AMPA-receptor patterns in macaque primary visual cortex.

Glutamate receptors mediate excitatory neurotransmission. A very prevalent type of glutamate receptor in the neocortex is the AMPA receptor (AMPAR). AMPARs mediate fast synaptic transmission and their functionality depends on the subunit composition. In primary visual cortex (area V1), the density and subunit composition of AMPARs differ among cortical layers and among cell types. The AMPARs expressed by the different types of inhibitory interneurons, which are crucial for network function, have not yet been characterized systematically. We investigated the distribution of AMPAR subunits in macaque V1 for three distinct subpopulations of inhibitory interneurons: parvalbumin-immunoreactive (PV-IR) interneurons, calbindin-immunoreactive (CB-IR) interneurons, and calretinin-immunoreactive (CR-IR) interneurons. We found that PV-IR cells, which have previously been identified as fast spiking, show high expression of the GluA2 and GluA3 subunits. In contrast, CB-IR and CR-IR cells, which tend to be intermediate spiking, show high expression of the GluA1 and GluA4 subunits. Thus, our data demonstrate that the expression of AMPARs divides inhibitory interneurons in macaque V1 into two categories that are compatible with existing classification methods based on calcium-binding proteins and firing behavior. Moreover, our findings suggest new approaches to target the different inhibitory interneuron classes pharmacologically in vivo.

Different glutamate receptors convey feedforward and recurrent processing in macaque V1.

Neurons in the primary visual cortex (V1) receive feedforward input from the thalamus, which shapes receptive-field properties. They additionally receive recurrent inputs via horizontal connections within V1 and feedback from higher visual areas that are thought to be important for conscious visual perception. Here, we investigated what roles different glutamate receptors play in conveying feedforward and recurrent inputs in macaque V1. As a measure of recurrent processing, we used figure-ground modulation (FGM), the increased activity of neurons representing figures compared with background, which depends on feedback from higher areas. We found that feedforward-driven activity was strongly reduced by the AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), whereas this drug had no effect on FGM. In contrast, blockers of the NMDA receptor reduced FGM, whereas their effect on visually driven activity varied with the subunit specificity of the drug. The NMDA receptor blocker 2-amino-5-phosphonovalerate (APV) caused a slight reduction of the visual response, whereas ifenprodil, which targets NMDA receptors containing the NMDA receptor NR2B subunit, increased the visual response. These findings demonstrate that glutamate receptors contribute differently to feedforward and recurrent processing in V1 and suggest ways to selectively disrupt recurrent processing so that its role in visual perception can be elucidated.

Flexible Polymer Electrodes for Stable Prosthetic Visual Perception in Mice.

Brain interfaces that can stimulate neurons, cause minimal damage, and work for a long time will be central for future neuroprosthetics. Here, the long-term performance of highly flexible, thin polyimide shanks with several small (<15 µm) electrodes during electrical microstimulation of the visual cortex, is reported. The electrodes exhibit a remarkable stability when several billions of electrical pulses are applied in vitro. When the devices are implanted in the primary visual cortex (area V1) of mice and the animals are trained to detect electrical microstimulation, it is found that the perceptual thresholds are 2-20 microamperes (µA), which is far below the maximal currents that the electrodes can withstand. The long-term functionality of the devices in vivo is excellent, with stable performance for up to more than a year and little damage to the brain tissue. These results demonstrate the potential of thin floating electrodes for the long-term restoration of lost sensory functions.

Chronic stability of a neuroprosthesis comprising multiple adjacent Utah arrays in monkeys.

Objective. Electrical stimulation of visual cortex via a neuroprosthesis induces the perception of dots of light ('phosphenes'), potentially allowing recognition of simple shapes even after decades of blindness. However, restoration of functional vision requires large numbers of electrodes, and chronic, clinical implantation of intracortical electrodes in the visual cortex has only been achieved using devices of up to 96 channels. We evaluated the efficacy and stability of a 1024-channel neuroprosthesis system in non-human primates (NHPs) over more than 3 years to assess its suitability for long-term vision restoration.Approach.We implanted 16 microelectrode arrays (Utah arrays) consisting of 8 × 8 electrodes with iridium oxide tips in the primary visual cortex (V1) and visual area 4 (V4) of two sighted macaques. We monitored the animals' health and measured electrode impedances and neuronal signal quality by calculating signal-to-noise ratios of visually driven neuronal activity, peak-to-peak voltages of the waveforms of action potentials, and the number of channels with high-amplitude signals. We delivered cortical microstimulation and determined the minimum current that could be perceived, monitoring the number of channels that successfully yielded phosphenes. We also examined the influence of the implant on a visual task after 2-3 years of implantation and determined the integrity of the brain tissue with a histological analysis 3-3.5 years post-implantation.Main results. The monkeys remained healthy throughout the implantation period and the device retained its mechanical integrity and electrical conductivity. However, we observed decreasing signal quality with time, declining numbers of phosphene-evoking electrodes, decreases in electrode impedances, and impaired performance on a visual task at visual field locations corresponding to implanted cortical regions. Current thresholds increased with time in one of the two animals. The histological analysis revealed encapsulation of arrays and cortical degeneration. Scanning electron microscopy on one array revealed degradation of IrOxcoating and higher impedances for electrodes with broken tips.Significance. Long-term implantation of a high-channel-count device in NHP visual cortex was accompanied by deformation of cortical tissue and decreased stimulation efficacy and signal quality over time. We conclude that improvements in device biocompatibility and/or refinement of implantation techniques are needed before future clinical use is feasible.

A Quantitative Comparison of Inhibitory Interneuron Size and Distribution between Mouse and Macaque V1, Using Calcium-Binding Proteins.

The mouse is a useful and popular model for studying of visual cortical function. To facilitate the translation of results from mice to primates, it is important to establish the extent of cortical organization equivalence between species and to identify possible differences. We focused on the different types of interneurons as defined by calcium-binding protein (CBP) expression in the layers of primary visual cortex (V1) in mouse and rhesus macaque. CBPs parvalbumin (PV), calbindin (CB), and calretinin (CR) provide a standard, largely nonoverlapping, labeling scheme in macaque, with preserved corresponding morphologies in mouse, despite a slightly higher overlap. Other protein markers, which are relevant in mouse, are not preserved in macaque. We fluorescently tagged CBPs in V1 of both species, using antibodies raised against preserved aminoacid sequences. Our data demonstrate important similarities between the expression patterns of interneuron classes in the different layers between rodents and primates. However, in macaque, expression of PV and CB is more abundant, CR expression is lower, and the laminar distribution of interneuron populations is more differentiated. Our results reveal an integrated view of interneuron types that provides a basis for translating results from rodents to primates, and suggest a reconciliation of previous results.