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. Author manuscript; available in PMC: 2020 Mar 2.
Published in final edited form as: Cell Rep. 2020 Feb 18;30(7):2360–2373.e5. doi: 10.1016/j.celrep.2020.01.055

Figure 5. Characterization of DLS PV Neurons.

Figure 5.

(A) Mapping of tdTomato-expressing cells in the medial and lateral part of DLS in the progeny of PV-Cre mice bred with Ai14 mice. Scale bar: 100 μm. Representative images for 3 independent animals. Scale bar: 100 μm.

(B) Quantifications of tdTomato-expressing cells in DLS along the rostrocaudal and mediolateral axis. Data (means ± SEM; n = 3 mice per group) were analyzed using paired two-tailed Student t test. *p < 0.05, medial versus lateral.

(C) Immunohistochemistry for PV and endogenous expression of tdTomato in the DLS of GAD2-Cre::Ai14, SST-Cre::Ai14 and PV-Cre::Ai14. Note the lack of overlap between PV and tdTomato in GAD2-Cre::Ai14 and SST-Cre:Ai14 mice. Representative images for 3 independent animals (single experiment). Scale bar: 25 μm.

(D) Quantifications of the overlap of tdTomato-expressing cells with PV immunpositive cells in the cingulate cortex, DLS, MS, nucleus accumbens shell and core, and caudate putamen medial and lateral of PV-Cre::Ai14. Data (means ± SEM; n = 3 mice per group).

(E) PV-Cre::TVA bigenic mice were injected with helper virus (AAV8-EF1a-FLEX-HB) followed by pseudotyped G-deleted rabies virus (EnvA-SADΔG-mCherry) in the DLS. Yellow arrowheads denote starter cells, which are positive for both GFP (helper) and mCherry (rabies). Representative images for 2 independent animals. Scale bar: 50 μm. Means ± SEM; n = 2 mice per group.

(F) Presynaptic partners were identified in the MS/DBN, dorsal subiculum, dorsal CA1, and dorsal and ventral CA3. Representative images for 2 independent animals. Scale bar: 100 μm. Means ± SEM; n = 2 mice per group.

(H–K) CA3 provides powerful synaptic input to PV neurons in DLS.

(H) Acute slices obtained from 6-week-old PV-Cre::Ai14 bigenic mice injected with CaMKII-ChR2 into CA3 were used for ex vivo slice electrophysiology. Solid yellow arrowhead indicates tdTomato-labeled PV neuron. Representative image for 4 independent animals. Scale bar: 10 μm.

(I) Light-evoked short latency (2 ± 0.7ms) excitatory postsynaptic currents (eEPSCs) were detected in PV and non-PV neurons following CA3 terminal activation. A light-evoked delayed (46 ± 5 ms) inhibitory current (eIPSC) was observed in a subset of non-PV neurons.

(J) Example recordings of blue-light-evoked inputs onto DLS neurons. Traces show synaptic currents (EPSCs: bottom, IPSCs: top) evoked in both PV- and non-PV-expressing neurons (red and black, respectively), and the 5-ms light pulse is indicated by a blue box.

(K) Average amplitude for both cell types. Data (means ± SEM; n = 6, 4 animals per group) were analyzed using unpaired Student two-tailed t test (detailed in Table S1).

(L) Immunohistochemistry for eYFP in the DLS, DBN, NAcc, BNST, LH, PVN, and SUM of PV-Cre mice injected with AAV5-DIO-ChR2-eYFP. Representative images for 3 independent animals (single experiment). Scale bar: 100 μm.