By Adalberto Merighi, Giorgio Carmignoto, C. Cuello
Research of the neural tissue provides exact and weird technical difficulties encountered in daily bench paintings. various books facing mobile and molecular protocols for normal use in mobile biology can be found, yet few are particularly dedicated completely to neurobiology. in addition, the "cross-talk" among researchers with diverse backgrounds, i.e. histologists, mobilephone and molecular biologists and physiologists, continues to be fairly tough, and extremely frequently one continues to be by some means "confined" to his or her personal particular box of craftsmanship by no means bold to discover "mysterious" lands except having the help of a giant laboratory past. the final suggestion past this undertaking used to be to place jointly the contributions from a few famous neuroscientists to provide a booklet that provides a survey of the main up-to-date suggestions for the learn of nerve cells. After many years spent doing examine within the neuroscience box, and having got a superb technical heritage in definite particular fields of neurobiology we've learned how tough is so that it will step right into a diversified know-how. This e-book endeavors to help in that objective.
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Extra resources for Cellular and Molecular Methods in Neuroscience Research
By contrast, at longer time intervals (45 min) it was extensively colocalized with syntaxin 6, a marker of the Trans-Golgi Network (TGN) and of the pericentriolar recycling endosome (Figure 2, C and C′). Studies in Brain Slices Examination of rat brain slices pulse-labeled with fluo-NT and chased for short periods of time (5–10 min) revealed a fairly diffuse distribution of the fluorescent label over nerve cell bodies and neuronal processes within regions documented to express high concentrations of NT1 receptors, such as the basal forebrain (10) and the ventral midbrain (11) (Figure 3A).
5. Rinse with oxygenated Ringer buffer for 5, 10, 15, 30, 45, or 60 minutes at 37°C. To control for nonspecific labeling, incubate additional slices in the presence of 100- to 1000-fold excess of 18 nonfluorescent probe or antagonist. 6. After rinsing, fix the slices for 30 minutes at room temperature with 4% PFA. 7. Immerse the slices overnight in the cryoprotectant solution, flatten on tissue chuck, snap freeze in isopentane at -60°C, and resection at 45 µm thickness in the plane of the slice on a freezing microtome.
1998. Internalization of D1 dopamine receptor in striatal neurons in vivo as evidence of action by dopamine agonists. J. Neurosci. 18:1650-1661. , P. Gaudreau, I. R. Cashman, and A. Beaudet. 1994. Synthesis of a biologically active fluorescent probe for labeling neurotensin receptors. J. Histochem. Cytochem. 42:755-763. , A. Alonso, D. Nouel, G. Gaudriault, M. -P. Vincent, and A. Beaudet. 1995. Somatodendritic internalization and perinuclear targeting of neurotensin in the mammalian brain. J. Neurosci.
Cellular and Molecular Methods in Neuroscience Research by Adalberto Merighi, Giorgio Carmignoto, C. Cuello