The binding of [ 3 H]ryanodine was determined in microsomal membrane preparations obtained from cultured cerebellar granule cells. A K D of 1 nM and a B max of 64 fmol/mg protein were calculated from saturation experiments. This binding was calcium dependent and maximum values were obtained at 100-3
Changes in the properties of developing glycine receptors in cultured mouse spinal neurons
✍ Scribed by Juan C. Tapia; Luis G. Aguayo
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 222 KB
- Volume
- 28
- Category
- Article
- ISSN
- 0887-4476
No coin nor oath required. For personal study only.
✦ Synopsis
We studied several neurophysiological properties of in vitro maturing glycine receptors in mouse spinal cord neurons cultured for various times: 3-7 days (early), 10-12 days (intermediate), and 17-24 days (mature), using whole-cell and gramicidin-perforated techniques. The glycine-activated Cl Ϫ conductance increased about 6-fold during in vitro development, and the current density increased from 177 Ϯ 42 pA/pF in early to 504 Ϯ 74 pA/pF in mature neurons. The sensitivity to glycine increased transiently from 39 Ϯ 2.8 µM in early neurons to 29 Ϯ 1 µM in intermediate neurons. Using whole-cell recordings, we found that E Cl did not change during development. With the gramicidin-perforated technique, on the other hand, E Cl shifted from Ϫ27 to Ϫ52 mV with development. Thus, immature neurons were depolarized by the activation of glycine receptors, whereas mature neurons were hyperpolarized. The current decayed (desensitized) during the application of 500 µM glycine. The decay was single exponential and the time constant increased from 2,212 Ϯ 139 msec in early neurons to 4,580 Ϯ 1,071 msec in mature neurons. Picrotoxin (10 µM) inhibited the current to a larger extent in early neurons (46 Ϯ 6% of control), and the sensitivity of these receptors to strychnine (IC 50 ) increased from 23 Ϯ 3 nM to 9 Ϯ 1 nM in mature neurons. In conclusion, several properties of spinal glycine receptors changed during in vitro neuronal maturation. This indicates that, similar to GABA A receptors, the functions of these receptors are developmentally regulated. These changes should affect the excitability of spinal neurons as well as other maturation processes.
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