AbstractThe lateral entorhinal cortex (LEC) provides a major cortical input to the hippocampal formation, equaling that of the medial entorhinal cortex (MEC). To understand the functional contributions made by LEC, basic knowledge of individual neurons, in the context of the intrinsic network, is needed. The aim of this study is to compare physiological and morphological properties of principal neurons in different LEC layers in postnatal rats. Using in vitro whole cell current‐clamp recordings from up to four post hoc morphologically identified neurons simultaneously, we established that principal neurons show layer specific physiological and morphological properties, similar to those reported previously in adults. Principal neurons in L(ayer) I, LII, and LIII have the majority of their dendrites and axonal collaterals alone in superficial layers. LV contains mainly pyramidal neurons with dendrites and axons extending throughout all layers. A minority of LV and all principal neurons in LVI are neurons with dendrites confined to deep layers and axons in superficial and deep layers. Physiologically, input resistances and time constants of LII neurons are lower and shorter, respectively, than those observed in LV neurons. Fifty‐four percent of LII neurons have sag potentials, resonance properties, and rebounds at the offset of hyperpolarizing current injection, whereas LIII and LVI neurons do not have any of these. LV neurons show prominent spike‐frequency adaptation and a decrease in spike amplitudes in response to strong depolarization. Despite the well‐developed interlaminar communication in LEC, the laminar differences in the biophysical and morphological properties of neurons suggest that their in vivo firing patterns and functions differ, similar to what is known for neurons in different MEC layers. © 2011 Wiley Periodicals, Inc.