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. 2021 May 17;12:2859. doi: 10.1038/s41467-021-22915-5

Fig. 8. LA connections, flatmaps of BLAa anterograde tracing, and BLAa functional recordings.

Fig. 8

a Summarized brain-wide connections of LA (ventromedial) neurons. For full list of abbreviations see Table 1. b Anterograde maps in top panels show weak projections from LA neurons to hippocampal regions SUBv and CA1v. Ellipse denotes location of CTb injection, which validates these sparse connections. c Retrograde map shows labeled cells in auditory cortical areas (AUD) following an LA retrograde tracer injection. Ellipse denotes location of PHAL injection in the primary auditory cortical area (AUDp), which shows strong anterograde label in LA validating AUD→LA connections. d LA receives strong input from ventromedial hypothalamic nucleus (VMH), as shown by the retrograde map. An AAV-RFP injection in VMH validates the VMH→LA connection showing strong label in LA, but also in BMAp (VMH→BMAp). The outputs of BLA.am (e), BLA.al (f), and BLA.ac (g) domains are represented at the macroscale level (gray matter region resolution) on a partial mouse brain flatmap75. The strength values of detected connections were binned into tertiles, and these are represented qualitatively as strong (maroon), moderate (red), weak (pink), and none (gray). h Longitudinal half of the entire CNS flatmap showing orientation and major brain divisions. The key to the color codes for connection strength is also shown. iq Functional characterization of synaptic innervation of projection-defined BLAa neurons. i, l, o Schematic drawings of experimental design and proposed circuitry. AAV-hSyn-ChR2-YFP (ChR2) is injected into PL (i), AId (l), or ILA (o) to label projection axons in BLA.am, BLA.al, and BLA.ac, respectively. Red retrobeads are injected into CP caudal dorsomedial (CPc.dm), CP caudal ventral (CPc.v), or CA3 to back-label projection neurons in BLA.am, BLA.al, or BLA.ac, respectively. Recordings are made from BLAa neurons labeled with red retrobeads while ChR2 labeled axons in BLA are stimulated. j, m, p LED pulse (5 ms)-evoked averaged response traces of an example neuron recorded at −70mV and +10 mV. Recordings were made in the presence of TTX and 4-AP to block polysynaptic inputs so that only monosynaptic excitatory responses are elicited. Average (± standard deviation) peak amplitude and latency of light pulse evoked EPSCs in red retrobead labeled BLA.am (k; 11/12 recorded neurons), BLA.al (n; 7/8 recorded neurons), and BLA.ac (q; 12/13 recorded neurons) neuron populations. Source data are provided as a Source Data file. r Left: excitatory and inhibitory responses (superimposed traces) evoked by blue light stimulation (5 ms). Right, responses from the same recorded cell in the presence of TTX and 4AP to show blockade of IPSPs.