title: Type I TARPs promote dendritic growth of early postnatal neocortical pyramidal cells in organotypic cultures.
abstract: The ionotropic α-amino-3-hydroxy-5-methyl-4-isoxazole propionate glutamate receptors (AMPARs) have been implicated in the establishment of dendritic architecture. The transmembrane AMPA receptor regulatory proteins (TARPs) regulate AMPAR function and trafficking into synaptic membranes. In the current study, we employ type I and type II TARPs to modulate expression levels and function of endogenous AMPARs and investigate in organotypic cultures (OTCs) of rat occipital cortex whether this influences neuronal differentiation. Our results show that in early development [5-10 days in vitro (DIV)] only the type I TARP γ-8 promotes pyramidal cell dendritic growth by increasing spontaneous calcium amplitude and GluA2/3 expression in soma and dendrites. Later in development (10-15 DIV), the type I TARPs γ-2, γ-3 and γ-8 promote dendritic growth, whereas γ-4 reduced dendritic growth. The type II TARPs failed to alter dendritic morphology. The TARP-induced dendritic growth was restricted to the apical dendrites of pyramidal cells and it did not affect interneurons. Moreover, we studied the effects of short hairpin RNA-induced knockdown of endogenous γ-8 and showed a reduction of dendritic complexity and amplitudes of spontaneous calcium transients. In addition, the cytoplasmic tail (CT) of γ-8 was required for dendritic growth. Single-cell calcium imaging showed that the γ-8 CT domain increases amplitude but not frequency of calcium transients, suggesting a regulatory mechanism involving the γ-8 CT domain in the postsynaptic compartment. Indeed, the effect of γ-8 overexpression was reversed by APV, indicating a contribution of NMDA receptors. Our results suggest that selected type I TARPs influence activity-dependent dendritogenesis of immature pyramidal neurons.
authors.affiliation: Developmental Neurobiology Group, Faculty for Biology and Biotechnology, Ruhr University Bochum, Bochum 44780, Germany.
authors.forename: Mohammad I K
authors.initials: MI
authors.lastname: Hamad
authors.forename: Alexander
authors.initials: A
authors.lastname: Jack
authors.forename: Oliver
authors.initials: O
authors.lastname: Klatt
authors.forename: Markus
authors.initials: M
authors.lastname: Lorkowski
authors.forename: Tobias
authors.initials: T
authors.lastname: Strasdeit
authors.forename: Sabine
authors.initials: S
authors.lastname: Kott
authors.forename: Charlotte
authors.initials: C
authors.lastname: Sager
authors.forename: Michael
authors.initials: M
authors.lastname: Hollmann
authors.forename: Petra
authors.initials: P
authors.lastname: Wahle
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015220:Calcium Channels / Q000737:chemistry / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D019579:Neocortex / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D009498:Neurotoxins / Q000633:toxicity; D009778:Occipital Lobe / Q000187:drug effects / Q000378:metabolism; D009924:Organ Culture Techniques; D017434:Protein Structure, Tertiary; D017966:Pyramidal Cells / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020318:Rats, Long-Evans; D018091:Receptors, AMPA / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: D015220:Calcium Channels; D009498:Neurotoxins; D012333:RNA, Messenger; D018091:Receptors, AMPA
keywords: Dendritogenesis; Glutamate receptors; Interneurons; Neocortex; Postnatal development; TARPs