AUTHOR=Meyer Jochen F. , Golshani Peyman , Smirnakis Stelios M. TITLE=The Effect of Single Pyramidal Neuron Firing Within Layer 2/3 and Layer 4 in Mouse V1 JOURNAL=Frontiers in Neural Circuits VOLUME=Volume 12 - 2018 YEAR=2018 URL=https://www.frontiersin.org/journals/neural-circuits/articles/10.3389/fncir.2018.00029 DOI=10.3389/fncir.2018.00029 ISSN=1662-5110 ABSTRACT=The influence of cortical cell spiking activity on nearby cells has been studied extensively in vitro. Less is known, however, about the impact of single cell firing on local cortical networks in vivo. In a pioneering study, Kwan et al. (Kwan 2012) reported that in mouse layer 2/3 (L2/3), under anesthesia, stimulating a single pyramidal cell recruits ~1.7% of neighboring pyramidal units. Here we employ two-photon calcium imaging in layer 2/3 of mouse V1, in conjunction with single-cell patch clamp stimulation in layer 2/3 or layer 4, to probe, in both the awake and lightly anesthetized states, how i) activating single L2/3 pyramidal neurons recruits neighboring units within L2/3 and from layer 4 (L4) to L2/3, and whether ii) activating single pyramidal neurons changes population activity in local circuit. To do this, it was essential to develop an algorithm capable of quantifying how sensitive the calcium signal is at detecting effectively recruited units (“followers”). This algorithm allowed us to estimate the chance of detecting a follower as a function of the probability that an epoch of stimulation elicits one extra action potential (AP) in the follower cell. Using this approach, we found only a small fraction (<0.75%) of L2/3 cells to be significantly activated within a radius of ~200 µm from a stimulated neighboring L2/3 pyramidal cell. This fraction did not change significantly in the awake versus the lightly anesthetized state, nor when stimulating L2/3 versus underlying L4 pyramidal neurons. These numbers are in general agreement with, though lower than, the percentage of neighboring cells (2.1%) reported by Kwan et al. to be activated upon stimulating single L2/3 pyramidal neurons under anesthesia (Kwan 2012). Interestingly, despite the small number of individual units found to be reliably driven, we did observe a modest but significant elevation in aggregate population responses compared to sham stimulation. This underscores the distributed impact that single cell stimulation has on neighboring microcircuit responses, revealing only a small minority of relatively strongly connected partners.