Modulation by serotonin reveals preferred recurrent excitatory connectivity in layer II of rat neocortex

We reported that in layer II pyramidal cells of rat somatosensory cortex, 10 μM serotonin (5-HT) alters miniature excitatory postsynaptic current frequency in a subset of cells (47%, "responders", RC; "non-responders", NC otherwise) via 5-HT2 receptors (5-HT2R) but in all pairs reduced evoked excitatory postsynaptic current amplitude by ~50% (Agahari FA, Stricker C. 2021. Serotonergic modulation of spontaneous and evoked transmitter release in layer II pyramidal cells of rat somatosensory cortex. Cereb Cortex. 31:1182-1200. https://doi.org/10.1093/cercor/bhaa285.) suggestive of preferential connectivity. We provide different lines of evidence that distinguish these subsets. First, after 5-HT exposure, changes in miniature excitatory postsynaptic current, spontaneous EPSC frequency, or whole-cell noise (σw) were restricted to postsynaptic cells in pairs (PO) and RC but absent in presynaptic (PR) and NC. Second, exposure caused a large change in holding current with a small variability in NC, but a small one with a large variability in PO/RC. In addition, ΔRin in PO/RC was larger than in PR/NC, with a negative correlation between ΔIhold and ΔRin in NC, a positive in PO, but none in RC. Third, an unbiased classifier identified most PO as RC and all PR as NC. Our data establish two distinct sets of pyramidal cells having a preferred connectivity from NC → RC. 5-HT2R-mediated modulation of transmitter release may likely reduce the signal-to-noise ratio in the ipsilateral but leave the output to the contralateral side unaffected.