Development and Function of Retinal Ganglion Cell Circuits

The function of our nervous system relies on specific patterns of synaptic connections between diverse neuronal cell types. My thesis research addressed how cell-type-specific patterns of connectivity emerge in the developing mouse retina and how they enable mature retinal neurons to detect specific sensory stimuli. Spike trains of the approximately 40 retinal ganglion cell (RGC) types in mammals encode specific features and events in the visual world, and are the sole source of visual information to the brain. Recent studies have begun to dissect the presynaptic circuits underlying diverse RGC light responses, but how cell-type-specific retinal circuits emerge during development is poorly understood. The first part of my dissertation explored the plasticity of the ON alpha (ONα-) RGC circuit. I found that developmental removal of the dominant excitatory input to ONα-RGCs triggers cell-type-specific rewiring, which precisely preserves ONα-RGCs’ characteristic light responses including high contrast sensitivity. Spiking neurons, including RGCs, typically encode sensory information by increasing firing rates in the presence of preferred stimuli. Suppressed-by-Contrast (SbC-) RGCs are unique in that they signal changes in luminance (i.e., contrast) by decreasing rather than increasing spiking. Taking advantages of mouse genetics, in the second part of my thesis, I characterized SbC-RGCs’ responses to complex stimuli and identified the synaptic mechanisms underlying their suppressive contrast encoding. Interestingly, I found that VGluT3-expressing amacrine cells (VG3-ACs) are dual transmitter neurons that release excitatory and inhibitory transmitters in a target-specific manner, and VG3-ACs specifically contributes to OFF inhibition to SbC-RGCs in response to small stimuli. Finally, using an intersectional transgenic approach, I preferentially labeled SbC-RGCs and mapped their central projections to explore the contribution of SbC-RGCs to vision

Layer-specific developmentally precise axon targeting of transient Suppressed-by-Contrast retinal ganglion cells (tSbC RGCs)

The mouse retina encodes diverse visual features in the spike trains of \u3e40 retinal ganglion cell (RGC) types. Each RGC type innervates a specific subset of the \u3e50 retinorecipient brain areas. Our catalog of RGC types and feature representations is nearing completion. Yet, we know little about where specific RGC types send their information. Furthermore, the developmental strategies by which RGC axons choose their targets and pattern their terminal arbors remain obscure. Here, we identify a genetic intersection

Genetically identified suppressed-by-contrast retinal ganglion cells reliably signal self-generated visual stimuli

Spike trains of retinal ganglion cells (RGCs) are the sole source of visual information to the brain; and understanding how the ∼20 RGC types in mammalian retinae respond to diverse visual features and events is fundamental to understanding vision. Suppressed-by-contrast (SbC) RGCs stand apart from all other RGC types in that they reduce rather than increase firing rates in response to light increments (ON) and decrements (OFF). Here, we genetically identify and morphologically characterize SbC-RGCs in mice, and target them for patch-clamp recordings under two-photon guidance. We find that strong ON inhibition (glycine > GABA) outweighs weak ON excitation, and that inhibition (glycine > GABA) coincides with decreases in excitation at light OFF. These input patterns explain the suppressive spike responses of SbC-RGCs, which are observed in dim and bright light conditions. Inhibition to SbC-RGC is driven by rectified receptive field subunits, leading us to hypothesize that SbC-RGCs could signal pattern-independent changes in the retinal image. Indeed, we find that shifts of random textures matching saccade-like eye movements in mice elicit robust inhibitory inputs and suppress spiking of SbC-RGCs over a wide range of texture contrasts and spatial frequencies. Similarly, stimuli based on kinematic analyses of mouse blinking consistently suppress SbC-RGC spiking. Receiver operating characteristics show that SbC-RGCs are reliable indicators of self-generated visual stimuli that may contribute to central processing of blinks and saccades. SIGNIFICANCE STATEMENT This study genetically identifies and morphologically characterizes suppressed-by-contrast retinal ganglion cells (SbC-RGCs) in mice. Targeted patch-clamp recordings from SbC-RGCs under two-photon guidance elucidate the synaptic mechanisms mediating spike suppression to contrast steps, and reveal that SbC-RGCs respond reliably to stimuli mimicking saccade-like eye movements and blinks. The similarity of responses to saccade-like eye movements and blinks suggests that SbC-RGCs may provide a unified signal for self-generated visual stimuli

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

Human adipose-derived stem cells (hADSCs) are easily isolated from fat tissue without ethical concerns, but differ in purity, pluripotency, differentiation ability, and stem cell marker expression, depending on the isolation method. We isolated hADSCs from a primary fat tissue solution using: (1) conventional culture, (2) a membrane filtration method, (3) a membrane migration method where the primary cell solution was permeated through membranes, adhered hADSCs were cultured, and hADSCs migrated out from the membranes. Expression of mesenchymal stem cell markers and pluripotency genes, and osteogenic differentiation were compared for hADSCs isolated by different methods using nylon mesh filter membranes with pore sizes ranging from 11 to 80 μm. hADSCs isolated by the membrane migration method had the highest MSC surface marker expression and efficient differentiation into osteoblasts. Osteogenic differentiation ability of hADSCs and MSC surface marker expression were correlated, but osteogenic differentiation ability and pluripotent gene expression were not

Task-Switching Performance Improvements After Tai Chi Chuan Training Are Associated With Greater Prefrontal Activation in Older Adults

Studies have shown that Tai Chi Chuan (TCC) training has benefits on task-switching ability. However, the neural correlates underlying the effects of TCC training on task-switching ability remain unclear. Using task-related functional magnetic resonance imaging (fMRI) with a numerical Stroop paradigm, we investigated changes of prefrontal brain activation and behavioral performance during task-switching before and after TCC training and examined the relationships between changes in brain activation and task-switching behavioral performance. Cognitively normal older adults were randomly assigned to either the TCC or control (CON) group. Over a 12-week period, the TCC group received three 60-min sessions of Yang-style TCC training weekly, whereas the CON group only received one telephone consultation biweekly and did not alter their life style. All participants underwent assessments of physical functions and neuropsychological functions of task-switching, and fMRI scans, before and after the intervention. Twenty-six (TCC, N = 16; CON, N = 10) participants completed the entire experimental procedure. We found significant group by time interaction effects on behavioral and brain activation measures. Specifically, the TCC group showed improved physical function, decreased errors on task-switching performance, and increased left superior frontal activation for Switch > Non-switch contrast from pre- to post-intervention, that were not seen in the CON group. Intriguingly, TCC participants with greater prefrontal activation increases in the switch condition from pre- to post-intervention presented greater reductions in task-switching errors. These findings suggest that TCC training could potentially provide benefits to some, although not all, older adults to enhance the function of their prefrontal activations during task-switching

Homeostatic plasticity in neural development

Abstract Throughout life, neural circuits change their connectivity, especially during development, when neurons frequently extend and retract dendrites and axons, and form and eliminate synapses. In spite of their changing connectivity, neural circuits maintain relatively constant activity levels. Neural circuits achieve functional stability by homeostatic plasticity, which equipoises intrinsic excitability and synaptic strength, balances network excitation and inhibition, and coordinates changes in circuit connectivity. Here, we review how diverse mechanisms of homeostatic plasticity stabilize activity in developing neural circuits

Target-Specific Glycinergic Transmission from VGluT3-Expressing Amacrine Cells Shapes Suppressive Contrast Responses in the Retina

Summary: Neurons that release more than one transmitter exist throughout the CNS. Yet, how these neurons deploy multiple transmitters and shape the function of specific circuits is not well understood. VGluT3-expressing amacrine cells (VG3-ACs) provide glutamatergic input to ganglion cells activated by contrast and motion. Using optogenetics, we find that VG3-ACs provide selective glycinergic input to a retinal ganglion cell type suppressed by contrast and motion (SbC-RGCs). Firing of SbC-RGCs is suppressed at light ON and OFF over a broad range of stimulus sizes. Anatomical circuit reconstructions reveal that VG3-ACs form inhibitory synapses preferentially on processes that link ON and OFF arbors of SbC-RGC dendrites. Removal of VG3-ACs from mature circuits reduces inhibition and attenuates spike suppression of SbC-RGCs in a contrast- and size-selective manner, illustrating the modularity of retinal circuits. VG3-ACs thus use dual transmitters in a target-specific manner and shape suppressive contrast responses in the retina by glycinergic transmission. : Tien et al. show that VG3-ACs deploy dual transmitters (glycine and glutamate) in a target-specific manner and form glycinergic synapses on the link processes connecting ON and OFF arbors of SbC-RGC dendrites. Cell-type-specific deletion in mature circuits reveals contrast- and size-selective influences of VG3-ACs on SbC-RGC responses

Homeostatic plasticity shapes the visual system’s first synapse

Retinal rod bipolar cells (RBCs) partially undergo programmed cell death triggering cell density-dependent plasticity. This study shows that increased removal of RBCs using genetic approaches causes dendrites of the remaining RBCs to expand and contact more rod photoreceptors while reducing connectivity with each

Discovery of a potent cyclooxygenase-2 inhibitor, S4, through docking-based pharmacophore screening, in vivo and in vitro estimations

Cyclooxygenase (COX; EC: 1.14.99.1), the key enzyme in prostaglandin production in the human body, is a major pharmacological target for developing anti-inflammatory agents. Nonsteroidal anti-inflammatory drugs exhibit anti-inflammatory and analgesic activities when inhibiting COX-2 but cause gastrointestinal toxicity and other side effects because of concurrent inhibition of COX-1. Thus, potent and safe inhibitors against COX-2 are urgently required. We constructed a novel docking-based pharmacophore model for screening selective COX-2 inhibitors and discovered compounds S1, S2, S3, and S4, which apparently inhibit COX-2. Particularly, S4 inhibits COX-2 in vitro and shows a potent anti-inflammatory effect in vivo without cytotoxicity. Molecular docking analyses revealed that S4 interacted satisfactorily with the active site of COX-2 but not with that of COX-1. This reveals that S4 more specifically inhibits COX-2 and has potential for application in developing anti-inflammatory and anticancer agents