आंदोलन के दौरान स्तनधारी न्यूरॉन्स के vivo शारीरिक प्रतिक्रिया में यों और neuronal आकारिकी, neurochemical phenotype और synaptic microcircuitry के साथ न्यूरॉन के शरीर क्रिया विज्ञान सहसंबंधी एक तकनीक वर्णित है.
The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor functions. Most investigations of sensorimotor mechanisms rely on either examination of neurons while an animal is static1,2 or record extracellular neuronal activity during a movement.3,4 While these studies have provided the fundamental background for sensorimotor function, they either do not evaluate functional information which occurs during a movement or are limited in their ability to fully characterize the anatomy, physiology and neurochemical phenotype of the neuron. A technique is shown here which allows extensive characterization of individual neurons during an in vivo movement. This technique can be used not only to study primary afferent neurons but also to characterize motoneurons and sensorimotor interneurons. Initially the response of a single neuron is recorded using electrophysiological methods during various movements of the mandible followed by determination of the receptive field for the neuron. A neuronal tracer is then intracellularly injected into the neuron and the brain is processed so that the neuron can be visualized with light, electron or confocal microscopy (Fig. 1). The detailed morphology of the characterized neuron is then reconstructed so that neuronal morphology can be correlated with the physiological response of the neuron (Figs. 2,3). In this communication important key details and tips for successful implementation of this technique are provided. Valuable additional information can be determined for the neuron under study by combining this method with other techniques. Retrograde neuronal labeling can be used to determine neurons with which the labeled neuron synapses; thus allowing detailed determination of neuronal circuitry. Immunocytochemistry can be combined with this method to examine neurotransmitters within the labeled neuron and to determine the chemical phenotypes of neurons with which the labeled neuron synapses. The labeled neuron can also be processed for electron microscopy to determine the ultrastructural features and microcircuitry of the labeled neuron. Overall this technique is a powerful method to thoroughly characterize neurons during in vivo movement thus allowing substantial insight into the role of the neuron in sensorimotor function.
यहाँ सचित्र विधि एक शक्तिशाली तकनीक है जो एकल न्यूरॉन्स के समारोह में महत्वपूर्ण अंतर्दृष्टि प्रदान करता है और कैसे व्यक्तिगत न्यूरॉन्स की प्रतिक्रिया neuronal सर्किट के लिए योगदान देता है 9 यह ज्ञान …
The authors have nothing to disclose.
मैं एंथोनी टेलर vivo intracellular रिकॉर्डिंग और intracellular धुंधला तकनीक के प्रारंभिक विकास के साथ मदद के लिए ब्राउन और दाऊद मैक्सवेल में प्रारंभिक प्रशिक्षण के लिए धन्यवाद. मैं एम. रजत collocalization मैक्रो के साथ मदद के लिए धन्यवाद. किसके साथ कई विद्वानों मैं इस तकनीक के विकास सहित, एम. आर. डोंगा Moritani, पी. लुओ, आर Ambalavanar में अंतर्दृष्टि प्रदान की सहयोग किया है. एनआईएच DE10132 अनुदान, DE15386 और RR017971 से काफी समर्थन के साथ इस तकनीक को विकसित किया गया था.
Name of reagent or equipment | Company | Catalogue number | Comments |
---|---|---|---|
electromagnetic vibrator | Ling Dynamic Systems | V101 | |
signal generator | Feedback Systems | PFG605 | capable of producing trapezoidal output signal |
electrode glass | Sutter Instruments | AF100-68-10 | with filament |
electrode puller | Sutter Instruments | Model P-2000 or P-80 | |
biotinamide | Vector Laboratories | SP-1120 | stored at 4°C |
Texas Red avidin DCS | Vector Laboratories | A-2016 | |
tetramethlyrhodamine | Molecular Probes | D-3308 | 3000 molecular weight, lysine fixable |
mouse anti-synaptophysin antibody | Chemicon | MAB5258 | |
fluorescent Nissl stain | Neurotrace, Molecular Probes | N-21480 | |
electrode tester | Winston Electronics | BL-1000-B | to measure electrode impedance |
electrometer | Axon Instruments | Axoprobe 1A, Axoclamp 2B |