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. 2019 May 29;39(22):4268–4281. doi: 10.1523/JNEUROSCI.2816-18.2019

Figure 4.

Figure 4.

Nicotine uncaging reveals nAChR functional upregulation in IPN neurons following chronic nicotine. a, PA-Nic was used to uncage nicotine with laser flash photolysis adjacent to IPN neurons in coronal brain slices of control and cNIC-treated mice. b, Variable IPN neuron morphology. Representative 2PLSM images of IPN neurons with complex (left image) and sparse (middle image) dendritic arbors are shown. Some neurons (middle image, boxed area exploded view in right image) have clear dendritic spines. c, Representative nicotine uncaging (50 ms, 2 mW, perisomatic stimulus) responses in an IPN neuron of a control and cNIC-treated mouse. Inset shows exploded view of the initial uncaging event. d, Summary data for all initial/first nicotine uncaging responses in IPN neurons of control [n = 11 cells; n = 3 mice (1 male/2 female)] and cNIC-treated [n = 11 cells; n = 4 mice (1 male/3 female)] mice. Cells/mice used were also used to derive data in c, e, f, and g. p Value, Unpaired t test. e, Summary time-series data for repeated (2 min interstimulus interval) nicotine uncaging responses in IPN neurons of control and cNIC-treated mice. Data at 0 min are the same data as in d but are replotted for clarity. Summary data from e for control (f) and cNIC-treated (g) mice are replotted on a normalized scale with individual cells shown in gray. h, Representative nicotine uncaging (50 ms, 2 mW; perisomatic stimulus) responses in IPN neurons from naive mice are shown for slices acutely treated with control ACSF or donepezil (1 μm; superfusion). Inset shows exploded view of the initial uncaging event. i, Summary data for all initial/first nicotine uncaging responses in control [n = 8 cells, 3 mice (2 male/1 female)] and donepezil-treated [n = 12 cells, 5 mice (3 male/2 female)] IPN neurons. p Value, Unpaired t test. j, Summary time-series data for repeated (2 min interstimulus interval) nicotine uncaging responses in control and donepezil-treated IPN neurons. Data at 0 min are the same data as in i but are replotted for clarity. k, l, Summary data from j for control (k) and donepezil-treated (l) slices are replotted on a normalized scale with individual cells shown in gray. m, Summary data for all initial/first nicotine uncaging responses in IPN neurons from cNIC-treated mice, where slices were either exposed to ACSF [control; n = 13 cells, 2 mice (1 male/1 female)] or donepezil [1 μm, superfusion; n = 11 cells, 2 mice (1 male/1 female)]. p Value, Unpaired t test.