Conditional depletion reveals temporal requirements for the oscillating transcription factor NHR-23/NR1F1 in C. elegans larval progression

Nematode molting is a remarkable process where the animals must essentially build a new epidermis underneath the old skin and then rapidly shed the old skin. The study of molting provides a gateway into the developmental program of many core cellular and physiological processes, such as oscillatory gene expression, coordinated intracellular trafficking, steroid hormone signaling, developmental timing, and extracellular remodeling. The nuclear hormone receptor NHR-23/NR1F1 is an important regulator of molting. Imaging and western blot time-courses revealed oscillatory NHR-23::GFP expression in the epithelium that closely followed the reported mRNA expression. Timed depletion experiments using the auxin-inducible degron system revealed that NHR-23/NR1F1 depletion early in a given larval stage caused animals to arrest with only weak molting defects, whereas later depletion resulted in highly penetrant severe molting and morphological defects. This larval arrest was independent of insulin signaling. Despite the weakly penetrant molting defects following early NHR-23/NR1F1 depletion, the epidermal barrier was defective suggesting that NHR-23/NR1F1 is necessary for establishing or maintaining this barrier. NHR-23/NR1F1 coordinates the expression of factors involved in molting, lipid transport/metabolism, and remodeling of the apical extracellular matrix. We propose that NHR-23/NR1F1 is a regulator in a recently discovered large-scale gene oscillatory network coordinating rhythmic skin regeneration.

Replicating Cortical Signatures May Open the Possibility for “Transplanting” Brain States via Brain Entrainment

Brain states, which correlate with specific motor, cognitive, and emotional states, may be monitored with noninvasive techniques such as electroencephalography (EEG) and magnetoencephalography (MEG) that measure macroscopic cortical activity manifested as oscillatory network dynamics. These rhythmic cortical signatures provide insight into the neuronal activity used to identify pathological cortical function in numerous neurological and psychiatric conditions. Sensory and transcranial stimulation, entraining the brain with specific brain rhythms, can effectively induce desired brain states (such as state of sleep or state of attention) correlated with such cortical rhythms. Because brain states have distinct neural correlates, it may be possible to induce a desired brain state by replicating these neural correlates through stimulation. To do so, we propose recording brain waves from a “donor” in a particular brain state using EEG/MEG to extract cortical signatures of the brain state. These cortical signatures would then be inverted and used to entrain the brain of a “recipient” via sensory or transcranial stimulation. We propose that brain states may thus be transferred between people by acquiring an associated cortical signature from a donor, which, following processing, may be applied to a recipient through sensory or transcranial stimulation. This technique may provide a novel and effective neuromodulation approach to the noninvasive, non-pharmacological treatment of a variety of psychiatric and neurological disorders for which current treatments are mostly limited to pharmacotherapeutic interventions.

Cortical spheroids display oscillatory network dynamics

Three-dimensional brain cultures can facilitate the study of central nervous system function and disease, and one of the most important components that they present is neuronal activity on a network level. Here we demonstrate network activity in rodent cortical spheroids while maintaining the networks intact in their 3D state. Networks developed by nine days in culture and became more complex over time. To measure network activity, we imaged neurons in rat and mouse spheroids labelled with a calcium indicator dye, and in mouse spheroids expressing GCaMP. Network activity was evident when we electrically stimulated spheroids, was abolished with glutamatergic blockade, and was altered by GABAergic blockade or partial glutamatergic blockade. We quantified correlations and distances between somas with micron-scale spatial resolution. Spheroids seeded at as few as 4,000 cells gave rise to emergent network events, including oscillations. These results are the first demonstration that self-assembled rat and mouse spheroids exhibit network activity consistent with in vivo network events. These results open the door to experiments on neuronal networks that require fewer animals and enable high throughput experiments on network-perturbing alterations in neurons and glia.

Cortical spheroids display oscillatory network dynamics

Three-dimensional brain cultures can facilitate the study of central nervous system function and disease, and one of the most important components that they present is neuronal activity on a network...

A proof of concept ‘phase zero’ study of neurodevelopment using brain organoid models with Vis/near-infrared spectroscopy and electrophysiology

Homeostatic control of neuronal excitability by modulation of synaptic inhibition (I) and excitation (E) of the principal neurons is important during brain maturation. The fundamental features of in-utero brain development, including local synaptic E–I ratio and bioenergetics, can be modeled by cerebral organoids (CO) that have exhibited highly regular nested oscillatory network events. Therefore, we evaluated a 'Phase Zero' clinical study platform combining broadband Vis/near-infrared(NIR) spectroscopy and electrophysiology with studying E–I ratio based on the spectral exponent of local field potentials and bioenergetics based on the activity of mitochondrial Cytochrome-C Oxidase (CCO). We found a significant effect of the age of the healthy controls iPSC CO from 23 days to 3 months on the CCO activity (chi-square (2, N = 10) = 20, p = 4.5400e−05), and spectral exponent between 30–50 Hz (chi-square (2, N = 16) = 13.88, p = 0.001). Also, a significant effect of drugs, choline (CHO), idebenone (IDB), R-alpha-lipoic acid plus acetyl-l-carnitine (LCLA), was found on the CCO activity (chi-square (3, N = 10) = 25.44, p = 1.2492e−05), spectral exponent between 1 and 20 Hz (chi-square (3, N = 16) = 43.5, p = 1.9273e−09) and 30–50 Hz (chi-square (3, N = 16) = 23.47, p = 3.2148e−05) in 34 days old CO from schizophrenia (SCZ) patients iPSC. We present the feasibility of a multimodal approach, combining electrophysiology and broadband Vis–NIR spectroscopy, to monitor neurodevelopment in brain organoid models that can complement traditional drug design approaches to test clinically meaningful hypotheses.