SARS-CoV-2 spike host cell surface exposure promoted by a COPI sorting inhibitor

Via an insufficient coat protein complex I (COPI) retrieval signal, the majority of SARS-CoV-2 spike (S) is resident in host early secretory organelles and a tiny amount is leaked out in cell surface. Only surface-exposed S can be recognized by B cell receptor (BCR) or anti-S therapeutic monoclonal antibodies (mAbs) that is the trigger step for B cell activation after S mRNA vaccination or infected cell clearance by S mAbs. Now, a drug strategy to promote S host surface exposure is absent. Here, we first combined structural and biochemical analysis to characterize S COPI sorting signals. A potent S COPI sorting inhibitor was then invented, evidently capable of promoting S surface exposure and facilitating infected cell clearance by S antibody-dependent cellular cytotoxicity (ADCC). Importantly, with the inhibitor as a probe, we revealed Omicron BA.1 S is less cell surface exposed than prototypes because of a constellation of S folding mutations, possibly corresponding to its ER chaperone association. Our findings not only suggest COPI is a druggable target against COVID-19, but also highlight SARS-CoV-2 evolution mechanism driven by S folding and trafficking mutations.

Application of Tetrode Technology for Analysis of Changes in Neural Excitability of Medial Vestibular Nucleus by Acute Arterial Hypotension / 대한평형의학회지

OBJECTIVES: Excitability o medial vestibular nucleus (MVN) in the brainstem can be affected by changes in the arterial blood pressure. Several animal studies have demonstrated that acute hypotension results in the alteration of multiunit activities and expression of cFos protein in the MVN. In the field of extracellular electrophysiological recording, tetrode technology and spike sorting algorithms can easily identify single unit activity from multiunit activities in the brain. However, detailed properties of electrophysiological changes in single unit of the MVN during acute hypotension have been unknown. METHODS: Therefore, we applied tetrode techniques and electrophysiological characterization methods to know the effect of acute hypotension on single unit activities of the MVN of rats. RESULTS: Two or 3 types of unit could be classified according to the morphology of spikes and firing properties of neurons. Acute hypotension elicited 4 types of changes in spontaneous firing of single unit in the MVN. Most of these neurons showed excitatory responses for about within 1 minute after the induction of acute hypotension and then returned to the baseline activity 10 minutes after the injection of sodium nitroprusside. There was also gradual increase in spontaneous firing in some units. In contrast small proportion of units showed rapid reduction of firing rate just after acute hypotension. CONCLUSIONS: Therefore, application of tetrode technology and spike sorting algorithms is another method for the monitoring of electrical activity of vestibular nuclear during acute hypotension.

Registro de actvvidad eléctrica en la retina deuna rata albina empleando una mmatriz de microelectrooos / Recording of Electrical Activity in the Retina of an Albino Rat Employing a Microelectrode Array

Las matrices de microelectrodos (MEA) son dispositivos que permiten la detección de potenciales de acción o espigas en poblaciones de células excitables, ofreciendo varias aplicaciones en el campo de las neurociencias y la biología. Este trabajo muestra un protocolo para el registro de espigas en una población de células ganglionares retinales empleando una matriz de microelectrodos. La retina de una rata albina fue extraída y preparada para ser estimulada in vitro con luz led blanca, con el fin de registrar sus espigas evocadas ante estos estímulos. Cada microelectrodo puede registrar espigas de más de una célula ganglionar, razón por la cual se determinó a qué célula pertenece cada espiga aplicando un procedimiento conocido como "clasificación de espigas". El trabajo permitió obtener el registro de un periodo de estimulación y otro de no estimulación, con el fin de representar los potenciales de acción evocados con luz y los espontáneos. Los registros fueron almacenados para visualizar las espigas de las células ganglionares y poder aplicar la herramienta de clasificación de espigas. De este modo, se almacenan los instantes de tiempo en los cuales cada célula ganglionar registrada generó potenciales de acción. Este trabajo conllevó al establecimiento de un protocolo de experimentación básico enfocado al uso de matrices MEA en el laboratorio de adquisición de potenciales extracelulares de la Universidad Antonio Nariño Sede Bogotá, no sólo para caracterizar los potenciales de acción de células ganglionares retinales, sino también para otro tipo de células que puedan ser estudiadas empleando matrices de microelectrodos.

The microelectrode arrays (MEA) are devices that allow the detection of action potentials or spikes in populations of excitable cells, offering a wide spectrum of applications in topics of Neurosciences and Biology. This work describes a protocol for recording of spikes in a population of retinal ganglion cells employing a microelectrode array. The retina of an albino rat was dissected and prepared to be stimulated in vitro with white led light and to record their evoked spikes. Each microelectrode can record spikes from more than a ganglion cell, for which it was necessary to determine which cell fires each spike applying a procedure known as spike sorting. The work allowed to obtain the recording of a stimulation period and another of non-stimulation, representing evoked and spontaneous action potentials. The recordings were saved, in order to visualize the action potentials of the ganglion cells detected and to apply a computational method for the spike sorting. In this way, it was saved the time stamps in which each action potential was fired by its respective cell. This work established a basic experimentation protocol focused to the use of MEA devices in the laboratory for acquisition of extracellular potentials at the Antonio Nariño University - Bogota Headquarters, not only for characterization of action potentials fired by retinal ganglion cells populations, but also for other kind of cells that can be studied employing MEA devices.

Waveform Sorting of Rabbit Retinal Ganglion Cell Activity Recorded with Multielectrode Array / 의학물리

Since the output of retina for visual stimulus is carried by neurons of very diverse functional properties, it is not adequate to use conventional single electrode for recording the retinal action potential. For this purpose, we used newly developed multichannel recording system for monitoring the simultaneous electrical activities of many neurons in a functioning piece of retina. Retinal action potentials are recorded with an extra-cellular planar array of 60 microelectrodes. In studying the collective activity of the ganglion cell population it is essential to recognize basic functional distinctions between individual neurons. Therefore, it is necessary to detect and to classify the action potential of each ganglion cell out of mixed signal. We programmed M-files with MATLAB for this sorting process. This processing is mandatory for further analysis, e.g. poststimulus time histogram (PSTH), auto-correlogram, and cross-correlogram. We established MATLAB based protocol for waveform classification and verified that this approach was effective as an initial spike sorting method.