Bidirectional Modulation of Intrinsic Excitability in Rat Prelimbic Cortex Neuronal Ensembles and Non-Ensembles after Operant Learning

Study Author Leslie Whitaker

Featured Paper of the Month – February 2018.

Learned associations between environmental stimuli and rewards drive learning and motivated behavior. These memories are thought to be encoded by alterations within specific patterns of sparsely distributed neurons called neuronal ensembles that are selectively activated by reward-predictive stimuli. Here we use the Fos promoter to identify strongly activated neuronal ensembles in rat prelimbic cortex (PLC) and assess altered intrinsic excitability following 10 days of operant food self-administration training. First, we selectively ablated Fos-expressing PLC neurons that were active during food self-administration. Selective removal of these neurons decreased food seeking. We then used male FosGFP transgenic rats to assess selective alterations of intrinsic excitability in Fos-expressing neuronal ensembles (FosGFP+) that were activated during food self-administration and compared these to alterations in less activated non-ensemble neurons (FosGFP-). Using whole cell recordings of pyramidal neurons in a brain slice preparation, we found that food self-administration increased excitability of FosGFP+ neurons and decreased excitability of FosGFP- neurons. Increased excitability of FosGFP+ neurons was driven by increased input resistance. Decreased excitability of FosGFP- neurons was driven by increased contribution of small conductance calcium-activated potassium (SK) channels. Injections of the specific SK channel antagonist apamin into PLC increased Fos expression but had no effect on food seeking. Overall, operant learning increased intrinsic excitability of PLC Fos-expressing neuronal ensembles that play a role in food seeking but decreased intrinsic excitability of Fos-negative non-ensembles.

Publication Information

Whitaker, Leslie R; Warren, Brandon L; Venniro, Marco; Harte, Tyler C; McPherson, Kylie B; Beidel, Jennifer; Bossert, Jennifer M; Shaham, Yavin; Bonci, Antonello; Hope, Bruce T

Bidirectional Modulation of Intrinsic Excitability in Rat Prelimbic Cortex Neuronal Ensembles and Non-Ensembles after Operant Learning. Journal Article

In: J Neurosci, vol. 37, no. 36, pp. 8845–8856, 2017, ISSN: 1529-2401 (Electronic); 0270-6474 (Linking).

Abstract | Links

@article{Whitaker:2017aa,

title = {Bidirectional Modulation of Intrinsic Excitability in Rat Prelimbic Cortex Neuronal Ensembles and Non-Ensembles after Operant Learning.},

author = {Leslie R Whitaker and Brandon L Warren and Marco Venniro and Tyler C Harte and Kylie B McPherson and Jennifer Beidel and Jennifer M Bossert and Yavin Shaham and Antonello Bonci and Bruce T Hope},

url = {https://www.ncbi.nlm.nih.gov/pubmed/28779019},

doi = {10.1523/JNEUROSCI.3761-16.2017},

issn = {1529-2401 (Electronic); 0270-6474 (Linking)},

year  = {2017},

date = {2017-09-06},

journal = {J Neurosci},

volume = {37},

number = {36},

pages = {8845--8856},

address = {Behavioral Neuroscience Research Branch and leslie.ramseychar64nih.gov.},

abstract = {Learned associations between environmental stimuli and rewards drive goal-directed learning and motivated behavior. These memories are thought to be encoded by alterations within specific patterns of sparsely distributed neurons called neuronal ensembles that are activated selectively by reward-predictive stimuli. Here, we use the Fos promoter to identify strongly activated neuronal ensembles in rat prelimbic cortex (PLC) and assess altered intrinsic excitability after 10 d of operant food self-administration training (1 h/d). First, we used the Daun02 inactivation procedure in male FosLacZ-transgenic rats to ablate selectively Fos-expressing PLC neurons that were active during operant food self-administration. Selective ablation of these neurons decreased food seeking. We then used male FosGFP-transgenic rats to assess selective alterations of intrinsic excitability in Fos-expressing neuronal ensembles (FosGFP(+)) that were activated during food self-administration and compared these with alterations in less activated non-ensemble neurons (FosGFP(-)). Using whole-cell recordings of layer V pyramidal neurons in an ex vivo brain slice preparation, we found that operant self-administration increased excitability of FosGFP(+) neurons and decreased excitability of FosGFP(-) neurons. Increased excitability of FosGFP(+) neurons was driven by increased steady-state input resistance. Decreased excitability of FosGFP(-) neurons was driven by increased contribution of small-conductance calcium-activated potassium (SK) channels. Injections of the specific SK channel antagonist apamin into PLC increased Fos expression but had no effect on food seeking. Overall, operant learning increased intrinsic excitability of PLC Fos-expressing neuronal ensembles that play a role in food seeking but decreased intrinsic excitability of Fos(-) non-ensembles.SIGNIFICANCE STATEMENT Prefrontal cortex activity plays a critical role in operant learning, but the underlying cellular mechanisms are unknown. Using the chemogenetic Daun02 inactivation procedure, we found that a small number of strongly activated Fos-expressing neuronal ensembles in rat PLC play an important role in learned operant food seeking. Using GFP expression to identify Fos-expressing layer V pyramidal neurons in prelimbic cortex (PLC) of FosGFP-transgenic rats, we found that operant food self-administration led to increased intrinsic excitability in the behaviorally relevant Fos-expressing neuronal ensembles, but decreased intrinsic excitability in Fos(-) neurons using distinct cellular mechanisms.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

Learned associations between environmental stimuli and rewards drive goal-directed learning and motivated behavior. These memories are thought to be encoded by alterations within specific patterns of sparsely distributed neurons called neuronal ensembles that are activated selectively by reward-predictive stimuli. Here, we use the Fos promoter to identify strongly activated neuronal ensembles in rat prelimbic cortex (PLC) and assess altered intrinsic excitability after 10 d of operant food self-administration training (1 h/d). First, we used the Daun02 inactivation procedure in male FosLacZ-transgenic rats to ablate selectively Fos-expressing PLC neurons that were active during operant food self-administration. Selective ablation of these neurons decreased food seeking. We then used male FosGFP-transgenic rats to assess selective alterations of intrinsic excitability in Fos-expressing neuronal ensembles (FosGFP(+)) that were activated during food self-administration and compared these with alterations in less activated non-ensemble neurons (FosGFP(-)). Using whole-cell recordings of layer V pyramidal neurons in an ex vivo brain slice preparation, we found that operant self-administration increased excitability of FosGFP(+) neurons and decreased excitability of FosGFP(-) neurons. Increased excitability of FosGFP(+) neurons was driven by increased steady-state input resistance. Decreased excitability of FosGFP(-) neurons was driven by increased contribution of small-conductance calcium-activated potassium (SK) channels. Injections of the specific SK channel antagonist apamin into PLC increased Fos expression but had no effect on food seeking. Overall, operant learning increased intrinsic excitability of PLC Fos-expressing neuronal ensembles that play a role in food seeking but decreased intrinsic excitability of Fos(-) non-ensembles.SIGNIFICANCE STATEMENT Prefrontal cortex activity plays a critical role in operant learning, but the underlying cellular mechanisms are unknown. Using the chemogenetic Daun02 inactivation procedure, we found that a small number of strongly activated Fos-expressing neuronal ensembles in rat PLC play an important role in learned operant food seeking. Using GFP expression to identify Fos-expressing layer V pyramidal neurons in prelimbic cortex (PLC) of FosGFP-transgenic rats, we found that operant food self-administration led to increased intrinsic excitability in the behaviorally relevant Fos-expressing neuronal ensembles, but decreased intrinsic excitability in Fos(-) neurons using distinct cellular mechanisms.