Introduction to Solid Supported Membrane Based Electrophysiology

The electrophysiological method we present is based on a solid supported membrane (SSM) composed of an octadecanethiol layer chemisorbed on a gold coated ...

Goethe University Frankfurt

Nociception generally evokes rapid withdrawal behavior in order to protect the tissue from harmful insults. Most nociceptive neurons responding to mechanical insults display highly branched dendrites, an anatomy shared by Caenorhabditis elegans FLP and PVD neurons, which mediate harsh touch responses. Although several primary molecular nociceptive sensors have been characterized, less is known about modulation and amplification of noxious signals within nociceptor neurons. First, we analyzed the FLP/PVD network by optogenetics and studied integration of signals from these cells in downstream interneurons. Second, we investigated which genes modulate PVD function, based on prior single-neuron mRNA profiling of PVD.

Optogenetic analysis of a nociceptor neuron and network reveals ion channels acting downstream of primary sensors.

Nociception &eacute;voque g&eacute;n&eacute;ralement le comportement de retrait rapide afin de prot&eacute;ger le tissu d&#39;insultes nuisibles. La plupart des neurones nociceptifs r&eacute;pondant aux agressions m&eacute;caniques affichent dendrites tr&egrave;s ramifi&eacute;es, une anatomie partag&eacute;e par Caenorhabditis elegans FLP et PVD neurones, qui n&eacute;gocient les r&eacute;ponses dures touch. Bien que plusieurs capteurs nociceptives mol&eacute;culaires principaux ont &eacute;t&eacute; caract&eacute;ris&eacute;s, on conna&icirc;t moins la modulation et l&#39;amplification des signaux nocifs dans les neurones nocicepteurs. Tout d&#39;abord, nous avons analys&eacute; le r&eacute;seau FLP/PVD par optogenetics et &eacute;tudi&eacute; l&#39;int&eacute;gration des signaux de ces cellules dans des interneurones en aval. Deuxi&egrave;mement, nous avons &eacute;tudi&eacute; Quels g&egrave;nes modulent la fonction PVD, bas&eacute;e sur le pr&eacute;c&eacute;dent single-neurone ARNm profilage des PVD.

Optogenetic analyse d&#39;un r&eacute;seau de neurones nocicepteurs et r&eacute;v&egrave;le des canaux ioniques agissant en aval de capteurs principaux.

Nocicezione evoca generalmente comportamento rapido ritiro al fine di proteggere il tessuto da insulti nocivi. La maggior parte dei neuroni nocicettivo risponde agli insulti meccanici display altamente ramificati dendriti, un&#39;anatomia condivisa da Caenorhabditis elegans FLP e PVD neuroni, che mediano risposte tocco duro. Anche se diversi primari sensori molecolari nocicettivo sono state caratterizzate, meno &egrave; conosciuto circa la modulazione e l&#39;amplificazione dei segnali nocivi all&#39;interno dei neuroni nocicettori. In primo luogo, abbiamo analizzato la rete FLP/PVD da optogenetics e studiato integrazione dei segnali da queste cellule in interneurons a valle. In secondo luogo, abbiamo studiato geni che modulano la funzione PVD, basato sul precedente singolo neurone mRNA profilatura di PVD.

Analisi Optogenetic di un neurone di nocicettori e rete rivelano scanalature dello ione, che agisce a valle di sensori primari.

C. elegans is used extensively as a model system in the neurosciences due to its well defined nervous system. However, the seeming simplicity of this nervous system in anatomical structure and neuronal connectivity, at least compared to higher animals, underlies a rich diversity of behaviors. The usefulness of the worm in genome-wide mutagenesis or RNAi screens, where thousands of strains are assessed for phenotype, emphasizes the need for computational methods for automated parameterization of generated behaviors. In addition, behaviors can be modulated upon external cues like temperature, O(subscript)2(/subscript) and CO(subscript)2(/subscript) concentrations, mechanosensory and chemosensory inputs. Different machine vision tools have been developed to aid researchers in their efforts to inventory and characterize defined behavioral "outputs". Here we aim at providing an overview of different worm-tracking packages or video analysis tools designed to quantify different aspects of locomotion such as the occurrence of directional changes (turns, omega bends), curvature of the sinusoidal shape (amplitude, body bend angles) and velocity (speed, backward or forward movement).

Keeping track of worm trackers.

C. elegans est largement utilis&eacute; comme un syst&egrave;me mod&egrave;le en neurosciences en raison de son syst&egrave;me nerveux bien d&eacute;fini. Cependant, la simplicit&eacute; apparente de ce syst&egrave;me nerveux dans la structure anatomique et la connectivit&eacute; neuronale, du moins par rapport &agrave; des animaux sup&eacute;rieurs, sous-tend une riche diversit&eacute; de comportements. L&#39;utilit&eacute; du ver dans le g&eacute;nome entier mutagenesis ou RNAi &eacute;crans, o&ugrave; des milliers de souches sont &eacute;valu&eacute;s pour le ph&eacute;notype, souligne la n&eacute;cessit&eacute; de m&eacute;thodes informatiques pour le param&eacute;trage automatique des comportements g&eacute;n&eacute;r&eacute;s. En outre, les comportements peuvent &ecirc;tre modul&eacute;s sur des facteurs externes comme la temp&eacute;rature, concentrations de O(subscript)2(/subscript) et CO(subscript)2(/subscript), m&eacute;canosensoriels et chimiosensorielles entr&eacute;es. Outils de vision de machine diff&eacute;rents ont &eacute;t&eacute; d&eacute;velopp&eacute;s pour aider les chercheurs dans leurs efforts &agrave; inventorier et &agrave; caract&eacute;riser le comportements d&eacute;finis &laquo; sorties &raquo;. Ici, nous visons &agrave; fournir un aper&ccedil;u des diff&eacute;rentes ver-suivi colis ou analyse vid&eacute;o outils con&ccedil;us pour quantifier les diff&eacute;rents aspects de la locomotion par exemple l&#39;apparition de changements de direction (virages, virages om&eacute;ga), courbure de la forme sinuso&iuml;dale (amplitude, angle de pliage des corps) et la vitesse (vitesse, mouvement vers l&#39;avant ou arri&egrave;re).

Suivi des traqueurs de ver.

C. elegans &egrave; ampiamente utilizzato come un sistema di modello in Neuroscienze grazie al suo sistema nervoso ben definito. Tuttavia, l&#39;apparente semplicit&agrave; di questo sistema nervoso nella struttura anatomica e connettivit&agrave; neuronale, almeno rispetto agli animali superiori, sottende una ricca diversit&agrave; di comportamenti. L&#39;utilit&agrave; del worm in genome-wide mutagenesi o schermi RNAi, dove migliaia di ceppi &egrave; valutati per fenotipo, sottolinea la necessit&agrave; di metodi computazionali per la parametrizzazione automatica dei comportamenti generati. Inoltre, comportamenti possono essere modulati su segnali esterni come temperatura, delle concentrazioni di O(subscript)2(/subscript) e CO(subscript)2(/subscript), che mechanosensory e ingressi chemosensory. Macchina diversa visione strumenti sono stati sviluppati per aiutare i ricercatori nel loro sforzo di inventario e caratterizzare il comportamento definite &quot;uscite&quot;. Qui miriamo a fornire una panoramica dei diversi pacchetti di worm-monitoraggio o analisi video strumenti progettati per quantificare i diversi aspetti della locomozione quali il verificarsi di cambiamenti direzionali (curve, curve di omega), curvatura della forma sinusoidale (ampiezza, angoli di curvatura del corpo) e velocit&agrave; (velocit&agrave;, movimento all&#39;indietro o in avanti).

Wagner Steuer Costa

Buchmann Institute for Molecular Life Sciences

Goethe University Frankfurt

Introduction to Solid Supported Membrane Based Electrophysiology

Wagner Steuer Costa

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Buchmann Institute for Molecular Life Sciences

Goethe University Frankfurt

Introduction to Solid Supported Membrane Based Electrophysiology

Buchmann Institute for Molecular Life Sciences