Short- and long-lasting consequences of in vivo nicotine treatment on hippocampal excitability Rachel E. Penton, Michael W. Quick, Robin A. J. Lester Supplementary Figure 1. Histogram showing the maximal fepsp slope for different periods of withdrawal. 4-week (sal-slices, n = 12; nic-slices, n = 20), 3-month (sal-slices, n = 17; nic-slices, n = 22), 6-month (sal-slices, n = 6; nic-slices, n = 10), 9-month (salslices, n = 4; nic-slices, n = 5). Data are presented as the mean ± s.e.m.
Supplementary Figure 2. A, B, Mean miniature EPSP frequency (A) and amplitude (B) in CA1 pyramidal cells recorded in the presence of 1µM TTX from 1-day withdrawn animals (sal-cells, n = 4; nic-cells, n = 4). C, Timecourse of the resting membrane potential during TTX wash-in (sal-cells, n = 5; nic-cells, n = 4). Error bars are only shown once per minute for clarity. Data are presented as mean ± s.e.m. 2
Supplementary Figure 3. A, Mean current-voltage (I-V) curves for 500 ms hyperpolarizing and subthreshold depolarizing pulses in CA1 pyramidal cells from 1-day withdrawn animals in the presence of CNQX + D-APV (sal-cells, n = 5; nic-cells, n = 4). B, Mean current-voltage (I-V) curves for 500 ms hyperpolarizing and subthreshold depolarizing pulses in CA1 pyramidal cells from 1-day withdrawn animals in the presence of TTX (sal-cells, n = 4; nic-cells, n = 4). Data are presented as the mean ± s.e.m. 3
Supplementary Figure 4. A, Diagram of DG isolated slice. The scalpel cut is indicated. B, Mean frequencies of spontaneously occurring spikes recorded from DG stratum granulosum in both intact (sal-slices, n = 8; nic-slices, n = 7) and DG isolated (sal-slices, n = 8; nic-slices, n = 7) slices from 1-day withdrawn animals. Data are presented as the mean ± s.e.m. DG isolated, slices with connections severed between CA3 and dentate gyrus. 4
Supplementary Figure 5. A, Top: Examples of orthodromically evoked suprathreshold fepsps recorded in stratum radiatum (top traces) and pspikes recorded in stratum pyramidale (bottom traces) from 9-month withdrawn slices (sal-slices: red ; nic-slices: blue). Bottom: Examples of antidromically evoked aspikes recorded in stratum pyramidale (bottom traces) from 9-month withdrawn slices (sal-slices: red ; nic-slices: blue). Stimulus artifacts have been removed for clarity and black triangles represent time of stimulus. B, Mean extracellular E-S curves from long-term withdrawn (4-week + 9- month) no CA3 slices (sal-slices, n = 15; nic-slices, n = 19). C, Mean I-O curves for aspikes from long-term withdrawn (4-week + 9-month) no CA3 slices (sal-slices, n = 7; nic-slices, n = 7). Data are presented as the mean ± s.e.m. no CA3, slices with the CA3 region removed. 5
Supplementary Figure 6. A, Mean extracellular E-S curves from saline-treated 1-day withdrawn animals in no DG + no CA3 slices (n = 8) in ACSF (closed symbols) and in the presence of 500 nm nicotine (open symbols). B, Mean extracellular E-S curves from nicotine-treated 1-day withdrawn animals in no DG + no CA3 slices (n = 6) in ACSF (closed symbols) and in the presence of 500 nm nicotine (open symbols). Data are presented as the mean ± s.e.m. no DG, slices with connections severed between CA3 and dentate gyrus. no CA3, slices with the CA3 region removed. 6
Supplementary Table 1 Treatment Saline Nicotine Number of neurons 22 25 RMP (mv) -68.91 ± 0.85-64.24 ± 0.79* R input (MΩ) 30.18 ± 1.35 34.28 ± 1.56 Sag ratio 0.84 ± 0.01 0.85 ± 0.01 mahp (mv) -4.37 ± 0.27-4.83 ± 0.34 Action potential threshold (mv) -53.89 ± 1.32-54.46 ± 0.87 Action potential height (mv) 76.88 ± 0.97 77.46 ± 0.95 Action potential half-width (ms) 0.97 ± 0.01 0.99 ± 0.01 fahp (mv) -61.53 ± 1.49-62.51 ± 0.91 Single-cell properties following 1-day of withdrawal from chronic nicotine or saline. Membrane properties and action potential characteristics measured by intracellular recordings from CA1 pyramidal cells in hippocampal slices prepared from saline- or nicotine-treated animals following 1-day of withdrawal. Values represent mean ± s.e.m. * P < 0.05. 7
Supplementary Table 2 Treatment Saline Nicotine Number of neurons 14 14 RMP (mv) -69.79 ± 1.59-67.14 ± 1.35 R input (MΩ) 31.21 ± 1.94 38.36 ± 2.70* Sag ratio 0.86 ± 0.01 0.87 ± 0.01 mahp (mv) -4.90 ± 0.34-5.30 ± 0.62 Action potential threshold (mv) -52.30 ± 1.87-54.71 ± 1.21 Action potential height (mv) 76.30 ± 1.47 77.80 ± 1.49 Action potential half-width (ms) 1.00 ± 0.02 1.02 ± 0.01 fahp (mv) -60.88 ± 1.88-63.06 ± 1.39 Single-cell properties following long-term withdrawal from chronic nicotine or saline in the presence of blockers of synaptic transmission. Membrane properties and action potential characteristics measured by intracellular recordings from CA1 pyramidal cells in the presence of blockers in hippocampal slices prepared from saline- or nicotine-treated animals following long-term withdrawal (4- week + 3-month). Values represent mean ± s.e.m. * P < 0.05. 8