A method for evoking neurotransmitter release without light stimulation has been developed and applied to a retinal slice preparation of the tiger salamander (Ambystoma Tigrum). This method utilizes a micropipette containing hyperosmotic levels of sucrose in Ringer, positioned within the inner plexiform layer (IPL) under visual control. Intermittent pressure (between 0.1 and 2 bars) applied to the pipette evoked release of neurotransmitters which were evaluated with whole-cell recording (WCR) techniques applied to cells in the ganglion cell layer. Pharmacological studies were used to characterize the properties of the hyperosmotic sucrose-evoked response (HSER) and in some cases, we compared the HSER with synaptic currents evoked by light stimulation. The HSER typically consisted of both inhibitory and excitatory components with a reversal potential in between that for chloride (approximately -60 mV) and non-specific cation channels (approximately 0 mV). Relatively pure inhibition or excitation could be revealed through pharmacological techniques by blocking the inhibition with picrotoxin/strychnine or by blocking the glutamatergic neurotransmission with D-AP7 (D-2-amino-7-phosphonoheptanoate) and NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxaline). A comparison of lightevoked responses (LER) and the HSER suggested that they activate the same pool of releasable neurotransmitter.
Bibliographical noteFunding Information:
This work was supported by NIH Grant EY03014 to R.F.M.
- Ganglion cell
- Hyperosmotic sucrose
- Transmitter release