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1 Neurophysiology and New Microscopy, Institut National de la Sante et de la Recherche Medicale U603, Centre National de la Recherche Scientifique FRE 2500, Ecole Superieure de Physique et Chimie Industrielles, Paris, France
2 Chromocell Corporation, North Brunswick, New Jersey, USA
3 Department of Human Anatomy and Genetics, University of Oxford, Oxford, United Kingdom
* To whom correspondence should be addressed. E-mail: etienne.audinat{at}univ-paris5.fr.
Previous studies have shown that layer V pyramidal neurons projecting either to sub-cortical structures or the contralateral cortex undergo different morphological and electrophysiological patterns of development during the first three postnatal weeks. To isolate the determinants of this differential maturation, we analyzed the gene expression and intrinsic membrane properties of layer V pyramidal neurons projecting either to the superior colliculus (SC cells) or the contralateral cortex (CC cells) by combining whole-cell recordings and single-cell RT-PCR in acute slices prepared from postnatal day (P) 5-7 or P21-30 old mice. Among the 24 genes tested, the calcium channel subunits 
1B and 1C, the protease Nexin 1 and the calcium-binding protein calbindin were differentially expressed in adult SC and CC cells and the potassium channel subunit Kv4.3 was expressed preferentially in CC cells at both stages of the development. Intrinsic membrane properties, including input resistance, amplitude of the hyperpolarization-activated current and action potential threshold, differed quantitatively between the two populations as early as from the first postnatal week and persisted throughout adulthood. However, the two cell types had similar regular action potential firing behaviors at all developmental stages. Surprisingly, when we increased the duration of anesthesia with ketamine/xylazine or pentobarbital prior to decapitation, a proportion of mature SC cells, but not CC cells, fired bursts of action potentials. Altogether these results indicate that the two populations of layer V pyramidal neurons start to differ already during the first postnatal week and exhibit different firing capabilities following anesthesia.
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