सी एलिगेंस Neuromuscular जंक्शन से विदारक और रिकॉर्डिंग
Summary
इलेक्ट्रोफिजियोलॉजी की सुलभ synapses के लिए आवेदन synaptic गतिविधि का एक quantifiable उपाय, synaptic म्यूटेंट विश्लेषण में उपयोगी प्रदान करता है. यह लेख एक विच्छेदन neuromuscular (NMJ) जंक्शनों को बेनकाब करने के लिए इस्तेमाल किया विधि का वर्णन<em> Caenorhabditis एलिगेंस (सी एलिगेंस)</em> और संक्षिप्त का उपयोग करता है जो इस तैयारी लागू किया जा सकता है की कुछ चर्चा.
Abstract
Neurotransmission is the process by which neurons transfer information via chemical signals to their post-synaptic targets, on a rapid time scale. This complex process requires the coordinated activity of many pre- and post-synaptic proteins to ensure appropriate synaptic connectivity, conduction of electrical signals, targeting and priming of secretory vesicles, calcium sensing, vesicle fusion, localization and function of postsynaptic receptors and finally, recycling mechanisms. As neuroscientists it is our goal to elucidate which proteins function in each of these steps and understand their mechanisms of action. Electrophysiological recordings from synapses provide a quantifiable read out of the underlying electrical events that occur during synaptic transmission. By combining this technique with the powerful array of molecular and genetic tools available to manipulate synaptic proteins in C. elegans, we can analyze the resulting functional changes in synaptic transmission.
The C. elegans NMJs formed between motor neurons and body wall muscles control locomotion, therefore, mutants with uncoordinated locomotory phenotypes (known as unc s) often perturb synaptic transmission at these synapses 1. Since unc mutants are maintained on a rich supply of a bacterial food source, they remain viable as long as they retain some pharyngeal pumping ability to ingest food. This, together with the fact that C. elegans exist as hermaphrodites, allows them to pass on mutant progeny without the need for elaborate mating behaviors. These attributes, coupled with our recent ability to record from the worms NMJs 2,3,7 make this an excellent model organism in which to address precisely how unc mutants impact neurotransmission.
The dissection method involves immobilizing adult worms using a cyanoacrylic glue in order to make an incision in the worm cuticle exposing the NMJs. Since C. elegans adults are only 1 mm in length the dissection is performed with the use of a dissecting microscope and requires excellent hand-eye coordination. NMJ recordings are made by whole-cell voltage clamping individual body wall muscle cells and neurotransmitter release can be evoked using a variety of stimulation protocols including electrical stimulation, light-activated channel-rhodopsin-mediated depolarization 4 and hyperosmotic saline, all of which will be briefly described.
Protocol
Discussion
आनुवंशिक मॉडल जीव सी एलिगेंस, आदर्श synaptic प्रसारण के अध्ययन के लिए अनुकूल है synaptic प्रोटीन जीन एन्कोडिंग में परिवर्तन के कार्यात्मक विश्लेषण के माध्यम से. यहाँ हम एक विच्छेदन तकनीक है कि सी. renders वर्ण?…
Acknowledgements
इस लेख JER के लिए NIH अनुदान द्वारा वित्त पोषित किया गया था. मैं cholinergic एक रिकॉर्डिंग के प्रदर्शनों की में प्रयुक्त मोटर न्यूरॉन्स में ट्रांसजेनिक चैनल rhodopsin व्यक्त कीड़े प्रदान करने के लिए डा. एलेक्स Gottschalk धन्यवाद देना चाहूंगा.
References
- Brenner, S. . Genetics. 77 (1), 71-71 (1974).
- Richmond, J. E., Davis, W. S., Jorgensen, E. M. . Nat Neurosci. 2 (11), 959-959 (1999).
- Richmond, J. E. . WormBook. 1, (2006).
- Liewald, J. F., Brauner, M., Stephens, G. J. . Nature methods. 5 (10), 895-895 (2008).
- Lockery, S. R., Goodman, M. B. . Methods in enzymology. 293, 201-201 (1998).
- Brockie, P. J., Mellem, J. E., Hills, T. . Neuron. 31 (4), 617-617 (2001).
- Richmond, J. E., Jorgensen, E. M. . Nat Neurosci. 2 (9), 791-791 (1999).
