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Rode Crowder posted an update 6 months, 3 weeks ago
The vagus nerve plays a pivotal role in communication between the brain and peripheral organs involved in the sensory detection and the autonomic control of visceral activity. While the lack of appropriate experimental techniques to manipulate the physiological activity of the vagus nerve has been a long-standing problem, recent advancements in optogenetic tools, including viral vectors and photostimulation devices, during the late 2010s have begun to overcome this technical hurdle. Furthermore, identifying promoters for expressing transgenes in a cell-type-specific subpopulation of vagal neurons enables the selective photoactivation of afferent/efferent vagal neurons and specific visceral organ-innervating vagal neurons. In this chapter, we describe recent optogenetic approaches to study vagus nerve physiology and describe how these approaches have provided novel findings on the roles of vagus nerve signals in the cardiac, respiratory, and gastrointestinal systems. Compared with studies of the central nervous system, there are still few insights into vagus nerve physiology. Further studies with optogenetic tools will be useful for understanding the fundamental characteristics of vagus nerve signals transferred throughout the body.Using an optogenetic approach, we analyzed a local neuron network of the respiratory center in the medulla of a brainstem-spinal cord preparation isolated from neonatal rat. We developed a transgenic (Tg) rat line in which Phox2b-positive cells expressed archaerhodopsin-3 (Arch) or one of the step-function channelrhodopsin variants (ChRFR) under the control of Phox2b promoter-enhancer regions. Then, in en bloc preparations from 0- to 2-day-old Tg neonatal rats, we analyzed membrane potential changes of medullary respiratory-related neurons in response to photostimulation of the rostral ventral medulla. The photostimulation-induced inhibition or facilitation of the respiratory rhythm in Arch-expressing or ChRFR-expressing Tg rat preparations, respectively. Selective photoactivation of Phox2b-positive neurons expressing ChRFR in the rostral ventrolateral medulla of a neonatal rat en bloc preparation induced membrane potential changes of respiratory-related neurons that were dependent on heterogeneous properties of synaptic connections in the respiratory center. We concluded that the optogenetic approach is a powerful method of verifying a hypothetical model of local networks among respiratory-related neurons in the rostral ventrolateral medulla of neonatal rat.The formation and maintenance of episodic memories are important for our daily life. Accumulating evidence from extensive studies with pharmacological, electrophysiological, and molecular biological approaches has shown that both entorhinal cortex (EC) and hippocampus (HPC) are crucial for the formation and recall of episodic memory. However, to further understand the neural mechanisms of episodic memory processes in the EC-HPC network, cell-type-specific manipulation of neural activity with high temporal resolution during memory process has become necessary. Recently, the technological innovation of optogenetics combined with pharmacological, molecular biological, and electrophysiological approaches has significantly advanced our understanding of the circuit mechanisms for learning and memory. Optogenetic techniques with transgenic mice and/or viral vectors enable us to manipulate the neural activity of specific cell populations as well as specific neural projections with millisecond-scale temporal control during animal behavior. Integrating optogenetics with drug-regulatable activity-dependent gene expression systems has identified memory engram cells, which are a subpopulation of cells that encode a specific episode. Finally, millisecond pulse stimulation of neural activity by optogenetics has further achieved (a) identification of synaptic connectivity between targeted pairs of neural populations, (b) cell-type-specific single-unit electrophysiological recordings, and (c) artificial induction and modification of synaptic plasticity in targeted synapses. In this chapter, we summarize technological and conceptual advancements in the field of neurobiology of learning and memory as revealed by optogenetic approaches in the rodent EC-HPC network for episodic memories.Neural circuit function is determined not only by anatomical connections but also by the strength and nature of the connections, that is functional or physiological connectivity. To elucidate functional connectivity, selective stimulation of presynaptic terminals of an identified neuronal population is crucial. However, in the central nervous system, intermingled input fibers make selective electrical stimulation impossible. With optogenetics, this becomes possible, and enables the comprehensive study of functional synaptic connections between an identified population of neurons and defined postsynaptic targets to determine the functional connectome. Degrasyn nmr By stimulating convergent synaptic inputs impinging on individual postsynaptic neurons, low frequency and small amplitude synaptic connections can be detected. Further, the optogenetic approach enables the measurement of cotransmission and its relative strength. Recently, optogenetic methods have been more widely used to study synaptic connectivity and revealed novel synaptic connections and revised connectivity of known projections. In this chapter, I focus on functional synaptic connectivity in the striatum, the main input structure of the basal ganglia, involved in the motivated behavior, cognition, and motor control, and its disruption in a range of neuropsychiatric disorders.Optogenetics, which relies on the use of photons to manipulate cellular and subcellular processes, has emerged as an important tool that has transformed several fields including neuroscience. Improvement of optogenetic topographies, together with integration with complementary tools such as electrophysiology, imaging, anatomical and behavioral analysis, facilitated this transformation. However, an inherent challenge associated with optogenetic manipulation of neurons in living organisms, such as rodents, is the requirement for implanting light-delivering optical fibers. This is partly because the current repertoires of light-sensitive opsins are activated only by visible light, which cannot effectively penetrate biological tissues. Insertion of optical fibers and subsequent photo-stimulation inherently damages brain tissue, and fiber tethering can constrain animal behavior. To overcome these technical limitations, we and other research groups recently developed minimally invasive “fiberless optogenetics,” which uses particles that can emit visible light through up-conversion luminescence in response to irradiation with tissue-penetrating near-infrared light.
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