शारीरिक, आंदोलन द्वारा संग्राहक न्यूरॉन्स के morphological और neurochemical विशेषता

Summary

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.

Abstract

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 static^1,2 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.

Protocol

1. पशु तैयारी Pentobarbital सोडियम (50mg/kg आईपी) और एक हीटिंग पैड पर जगह के साथ चूहे anesthetize. जानवरों की क़ैंची के साथ पीछे खोपड़ी overlying त्वचा दाढ़ी. विश्वास दिलाता हूं कि संज्ञाहरण के स्तर के एक शल्य चिकित्सा स्तर एक व?…

Discussion

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 …

Materials

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