Materials
| Name | Company | Catalog Number | Comments |
| (1) Tissue culture incubator (H) | |||
| (1) Heat-resistant plastic enclosure (A) | (delrin or comparable) | ||
| (1) Plastic frame for 35mm GBM lid (M) | (for affixing membrane to petri dish) | ||
| (1-2) linear ft. Clear PFTE (Teflon) | membrane material | ||
| (4) Brass thumb screws | |||
| (2) Small 10kOhm heatsink resistors | used as heating elements inside stage-top incubator enclosure | ||
| (1) Transformer | supply of 9V current to resistors | ||
| (1) Terrarium temperature controller and probe | thermostatic regulation of power to heatsink resistors via transformer | ||
| (1) 2000ml Erlenmeyer flask (K) | muffler for vibration suppression in hose before stage-top incubator | ||
| (1) 1/8" sorbothane sheet | gasket material for base of stage-top incubator | ||
| (1) Airtight ABS (or comparable) plastic box (J) | housing for aquarium pump | ||
| (1) Aquarium pump (I) | |||
| (1) Small computer cooling fan | |||
| (1) Plastic enclosure for cooling fan (L) | |||
| (1) 9V transformer for computer cooling fan | |||
| (20-30ft) Nalgene or silicone hose (F,G,N) | connections between tissue culture and stage-top incubators; moisture traps | ||
| (2) Plastic stopcocks (O) | to open and close air flow before and after stage-top incubator | ||
| (4) Barbed brass hose connectors | for hose connections to/from stage-top incubator and aquarium pump enclosure | ||
| (2) Brass quick couplers | for hose connections to/from stage-top incubator | ||
| (2) Brass quick couplers | for hose connections to/from stage-top incubator | ||
| Table 1. Stage-top incubator parts: | |||
| Poly-D-lysine | Sigma-Aldrich | P7280-5MG | |
| laminin | Roche Group | 11243217001 | |
| 35 mm glass-bottom dishes | MatTek Corp. | P35GC-0-14-C | |
| DMEM | Life Technologies | 10567 | |
| B27 | Life Technologies | 17504-044 | |
| Glutamax | Life Technologies | 35050 | |
| Lipid-rich BSA | Life Technologies | 11020-021 | |
| L-Asparagine | Sigma-Aldrich | P0380-100G | |
| L-Proline | Sigma-Aldrich | A8381-100G | |
| Vitamin B-12 | Sigma-Aldrich | V2876-100MG | |
| 5-HT | Sigma-Aldrich | H9523-25MG | |
| 8-OH-DPAT | Sigma-Aldrich | H8520-25MG | |
| Dopamine | Sigma-Aldrich | H8502-5G | |
| Bromocriptine | Sigma-Aldrich | B2134-25MG | |
| SKF38393 | Sigma-Aldrich | D047-100MG | |
References
- Ligon, L. A., Steward, O. Movement of mitochondria in the axons and dendrites of cultured hippocampal neurons. J Comp Neurol. 427, 340-350 (2000).
- Miller, K. E., Sheetz, M. P. Direct evidence for coherent low velocity axonal transport of mitochondria. J Cell Biol. 173, 373-381 (2006).
- Macaskill, A. F. Miro1 is a calcium sensor for glutamate receptor-dependent localization of mitochondria at synapses. Neuron. 61, 541-555 (2009).
- Morris, R. L., Hollenbeck, P. J. Axonal transport of mitochondria along microtubules and F-actin in living vertebrate neurons. J Cell Biol. 131, 1315-1326 (1995).
- Rintoul, G. L., Filiano, A. J., Brocard, J. B., Kress, G. J., Reynolds, I. J. Glutamate decreases mitochondrial size and movement in primary forebrain neurons. J Neurosci. 23, 7881-7888 (2003).
- Crawley, J. N. Current protocols in neuroscience. , J. Wiley. (1999).
- Fedoroff, S., Richardson, A. Protocols for neural cell culture. , 3rd edn, Humana Press. (2001).
- Chen, S., Owens, G. C., Crossin, K. L., Edelman, D. B. Serotonin stimulates mitochondrial transport in hippocampal neurons. Mol Cell Neurosci. 36, 472-483 (2007).
- Chen, S., Owens, G. C., Edelman, D. B. Dopamine inhibits mitochondrial motility in hippocampal neurons. PLoS One. 3, e2804-e2804 (2008).
- Chen, S., Owens, G. C., Makarenkova, H., Edelman, D. B. HDAC6 regulates mitochondrial transport in hippocampal neurons. PLoS One. , e10848-e10848 (2010).
- Curran, M. A., Kaiser, S. M., Achacoso, P. L., Nolan, G. P. Efficient transduction of nondividing cells by optimized feline immunodeficiency virus vectors. Mol Ther. 1, 31-38 (2000).
