Interneurons transiently express the ERG K+ channels during development of mouse spinal networks in vitro.
During spinal cord maturation neuronal excitability gradually differentiates to meet different functional demands. Spontaneous activity, appearing early during spinal development, is regulated by the expression pattern of ion channels in individual neurons. While emerging excitability of embryonic motoneurons has been widely investigated, little is known about that of spinal interneurons. Voltage-dependent K+ channels are a heterogeneous class of ion channels that accomplish several functions. Recently voltage-dependent K+ channels encoded by erg subfamily genes (ERG channels) were shown to modulate excitability in immature neurons of mouse and quail. We investigated the expression of ERG channels in immature spinal interneurons, using organotypic embryonic cultures of mouse spinal cord after 1 and 2 weeks of development in vitro. We report here that all the genes of the erg family known so far (erg1a, erg1b, erg2, erg3) are expressed in embryonic spinal cultures. We demonstrate for the first time that three ERG proteins (ERG1A, ERG2 and ERG3) are co-expressed in the same neuronal population, and display a spatio-temporal distribution in the spinal slices. ERG immuno-positive cells, representing mainly GABAergic interneurons, were present in large numbers at early stages of development, while declining later, with a ventral to dorsal gradient. Patch clamp recordings confirmed these data, showing that ventral interneurons expressed functional ERG currents only transiently. Similar expression of the erg genes was observed at comparable ages in vivo. The role of ERG currents in regulating neuronal excitability during the earliest phases of spinal circuitry development will be examined in future studies.
Furlan
F
F
Physiology and Pathology Department, Center for Neuroscience B.R.A.I.N., Psychology Faculty, University of Trieste, via Sant'Anastasio 12, 34134, Trieste, Italy.
Guasti
L
L
Avossa
D
D
Becchetti
A
A
Cilia
E
E
Ballerini
L
L
Arcangeli
A
A
D000818:Animals; D004622:Embryo, Mammalian; D051638:Ether-A-Go-Go Potassium Channels / Q000096:biosynthesis* / Q000235:genetics; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017403:In Situ Hybridization; D007395:Interneurons / Q000378:metabolism*; D051379:Mice; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000196:embryology* / Q000378:metabolism
D051638:Ether-A-Go-Go Potassium Channels; D020033:Protein Isoforms