By Richard C. Rayne
Birkbeck collage, U.K. Bench-side significant other for researchers who search to spot, localize, or degree neurotransmitters and/or to spot websites of neurotransmitter motion. 36 participants, 6 U.S. broad references. DNLM: Neurotransmitters-physiology.
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Additional resources for Neurotransmitter Methods (Methods in Molecular Biology)
J Neuroscl Meth. 20, 83-90. 13. Kauppinen, R. A. and Nicholls, D. G. (1986) Synaptosomal bioenergettcs, the role of glycolysis, pyruvate oxidation and responses to hypoglycaemla. Eur J. Blochem. 158,159-165. 14. , Ghijsen, W. E. J. , and Lopes da Silva, F. H. (1994) Presynapttc plastictty* the regulation of Ca2+-dependent transmitter release Prog. Neurobzol 42,539-574. 15. McGeer, P. , Eccles, J. , and McGeer, E. G. (1987) Molecular Neurobiology the Mammalian Brain. Plenum, New York. of Synaptosomes 47 16.
Ca2+-free aCSF: the same as item 3 (high-Ca2+ aCSF) except add water mstead of CaCl, stock. 05 mA4. Stored at 4“C, the solution is stable for 2-3 d. 5. V, Alphen a/d Ryn, the Netherlands) and dinonylphthalate (Merck, Amsterdam, the Netherlands) at 45:55% (v/v). 3. 1, Preparation of Percoll Gradients (On Day of Use) 1. Add 2 mL of 23% Percoll/sucrose solution to each of three or four open-top, ultra-clear centrifuge tubes (see Note 2). 2. Using a Pasteur pipet, carefully layer 6 mL of 10% Percolhsucrose solutron onto the 23% PercolYsucrose solution in each tube.
The cell membrane should be smooth, clearly visible, and “phase-bright” under phase contrast mtcroscopy, the soma showing a bluish-green tinge with the dendrites more blue. Cells meeting these criteria generally then fulfill the electrophystologtcal criteria. Unhealthy cells have somas spotted with dark, granular patches or a uniform dark appearance For isolated dorsal raphe neurons, the electrophystological criteria mclude membrane potentials more negative than -55 mV and the ability to fire regular action potentials of brief duration that overshoot 0 mV (see Fig.