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. Author manuscript; available in PMC: 2019 Feb 7.
Published in final edited form as: Neuron. 2018 Jan 31;97(3):670–683.e6. doi: 10.1016/j.neuron.2018.01.016

Figure 5. Separate populations of vCA1 neurons project to the Basal Amygdala and Lateral Hypothalamus.

Figure 5

(A) Anterograde tracing of vCA1 axon terminals. CamKII-ChR2-eYFP virus was injected into vCA1 (left), and ChR2-eYFP axon terminals were visualized in the BA (middle) and LHA subfields (right).

(B) Average fluorescence of ChR2-eYFP terminal fields were similar between BA and LHA fields (paired t-test, t(4)=1.886, p=0.13, Nmice=5)

(C) In vitro slice recordings of BA cells in ChR2-eYFP vCA1 injected mice. 5ms 473nm light pulses elicited large optical EPSC (mean onset latency= 0.93 ms +/− 0.12 SEM; purple example trace, middle bar graph, Ncells=10), which were abolished by APV/NBQX infusion (black example trace; right bar graph Mann Whitney U=0.00, p<0.05, Ncells=4)

(D) In vitro slice recordings of LHA cells in ChR2-eYFP vCA1 injected mice. 5ms 473nm light pulses elicited large optical EPSC (mean onset latency= 1.56 ms +/− 0.09 SEM; purple example trace, middle bar graph, Ncells=10), which were abolished by APV/NBQX infusion (black example trace; right bar graph Mann Whitney U=0.00, p<0.01, Ncells=5)

(E) Ctb retrograde labeling of vCA1 projections to the BA (Ctb-555) and LHA (Ctb-488). Left, representative images of CTB injection sites in the BA (bottom) and LHA (top). Right, representative image of retrogradely labeled vCA1 neurons projecting to BA (red cells), LHA (green cells) or both (yellow cells, white arrows).

(F) Quantification of retrogradely labeled vCA1 neurons. vCA1-BA and LHA projecting neurons are largely non-overlapping as only ~3% of counted cells were dual-labeled (paired t-tests BA or LHA vs Dual, BA vs Dual t(10)=4.309, LHA vs Dual t(10)=5.832, p<0.01 for both).

(G) Left diagram, CA1 deep and superficial lamination. Right, cumulative distribution of vCA1-BA (red line) or vCA1-LHA (green line) labeled neuron distances from the inner radial border. The vCA1-LHA neuron distance distribution was significantly right shifted relative to vCA1-BA neurons, indicating that LHA neurons are organized deeper in the CA1 pyramidal layer (KS test, p<0.0001, KS stat=0.2999).

All data error bars represent mean +/− SEM