Spontaneous Auditory Neuron Activity

Shreya Sriram ,
1 George Ordiway 1 & Jason Tait Sanchez1,2,3

1Roxelyn and Richard Department of Communication Sciences and Disorders, 2Knowles Hearing Research Center, 3Department of Neurobiology, Northwestern University, Evanston, IL. USA

Abstract NMc sEPSCs AMPA-R Mediated sEPSCs

Synaptic connections between neurons is typically established through AMPA-R & GABAA-R AMPA-R Mediated GABAA-R Mediated AMPA-R Mediated AMPA-R Blocker Washout
spontaneous activity in vertebrates. Spontaneous activity is the occurrence of A Mediated sEPSCs B sEPSCs C sEPSCs A sEPSCs B (+CNQX) C (-CNQX)
neuronal events – such as intrinsic action potentials (APs) or postsynaptic
currents – in the absence of external stimuli. Generally, when a neuron is
isolated in a brainstem slice, spontaneous activity is not seen because
physiological conditions are not applicable. However, we have previously
reported spontaneous intrinsic AP activity in a subgroup of neurons in the
avian auditory brainstem (termed NMc). The goal of this project aimed to +PTX & AP-V +CNQX & AP-V +PTX & AP-V +PTX & AP-V +PTX & AP-V

characterize spontaneous synaptic activity of NMc neurons. We identified 0

spontaneous excitatory postsynaptic currents (sEPSCs) that were mediated
by ionotropic AMPA receptors (AMPA-Rs) and GABAA receptors (GABAA-Rs).
D D

Amplitude (pA)
Background

sEPSC Frequency (#)

A B

Figure 4: AMPA-R mediated sEPSCs from a NMc neuron. Example traces acquired before (A),
during (B), and after (C) bath application of CNQX (AMPA-R blocker). (B) Average amplitude before
and after AMPA-R blocker. Error bars = standard error of the mean.

GABAA-R Mediated sEPSCs

E
GABAA-R Mediated GABAA-R Blocker Washout
A B C
sEPSC Amplitude (pA)
sEPSCs (+PTX) (-PTX)

C Glutamate GABA
+CNQX & AP-V +CNQX & AP-V +CNQX & AP-V

Glu Glu Glu Glu GABA GABA

Amplitude (pA)
AMPA NMDA GABAA Figure 2. Spontaneous EPSCs from NMc neurons. (A) Example traces of sEPSCs without blocking
AMPA-Rs and GABAA-Rs. (B) AMPA-R mediated sEPSCs or (C) GABAA-R mediated sEPSCs. Violin
Receptor Receptor Receptor plots comparing sEPSCs frequency (D) or sEPSCs amplitude (E) during the three conditions .

sEPSCs Correlations
Figure 1. Structure and function of NMc neurons. (A) NMc neurons receive multiple synaptic
inputs onto their dendrites and soma, unlike traditional adendritic NM neurons which receive 1-2
A B
sEPSC Amplitude (pA)
sEPSC Frequency (#)

large Endbulb of Held synapses. (B) NMc neurons are more excitable, have a lower threshold for AP Figure 5: GABAA-R mediated sEPSCs from a NMc neuron. Example traces acquired before (A),
firing and are more depolarized compared to traditional NM neurons. (C) Schematic of the during (B), and after (C) bath application of PTX (GABAA-R blocker). (B) Average amplitude before and
neurotransmitters glutamate and GABA, and the postsynaptic receptors they bind to. Figure 1 (A & after GABAA-R blocker. Error bars = standard error of the mean.
B) adopted from Hong and Sanchez (2018), Journal of Experimental Neuroscience.

Conclusions
Method
1. sEPSCs of NMc neurons are mediated by AMPA-Rs and GABAA-Rs.
Whole-cell voltage clamp recordings were obtained from domesticated
C
sEPSC Frequency (#)

chickens at embryonic (E) days E19-E21. Spontaneous postsynaptic

excitatory currents (sEPSCs) were recorded by voltage clamping neurons
near their resting membrane potential (-60 mV) for 2 minutes. Synaptic
transmission through AMPA-Rs and GABAA-Rs were blocked depending on
which synaptic currents were isolated and studied. sEPSC properties were
recorded and analyzed before, during, and after bath application of CNQX
(AMPA-R blocker), or PTX (GABAA-R blocker). NMDA-Rs were blocked with
AP-V to isolate the other currents. Data reported are from 19 NMc neurons.
Funding: NIH, NIDCD R01 DC017167 & R03 DC013841 (JTS)
School of Communication and The Knowles Hearing Research Center, Northwestern University
2. Frequency and amplitude of sEPSCs were not correlated with low voltage
activated potassium currents (LVAK+), suggesting that spontaneous
synaptic activity is not related to intrinsic excitability.
3. sEPSC frequency and amplitude were correlated, suggesting that more
synaptic events have larger average amplitudes.
sEPSC Amplitude (pA)
Figure 3. Correlations of sEPSC frequency, amplitude and low voltage activated potassium 4. Spontaneous AMPA and GABA neurotransmission suggests that NMc
current (LVAK+). The relationship of sEPSC frequency (A) and amplitude (B) as a function of LVAK+ synaptic inputs are complex and specialized.
was not significant. (C) The relationship between sEPSC frequency and amplitude was significant.