Evolutionary insights into T-type Ca2+ channel structure, function, and ion selectivity from the <i>Trichoplax adhaerens</i> homologue

Abstract

Four-domain voltage-gated Ca²⁺ (Ca_v) channels play fundamental roles in the nervous system, but little is known about when or how their unique properties and cellular roles evolved. Of the three types of metazoan Ca_v channels, Ca_v1 (L-type), Ca_v2 (P/Q-, N- and R-type) and Ca_v3 (T-type), Ca_v3 channels are optimized for regulating cellular excitability because of their fast kinetics and low activation voltages. These same properties permit Ca_v3 channels to drive low-threshold exocytosis in select neurons and neurosecretory cells. Here, we characterize the single T-type calcium channel from <i>Trichoplax adhaerens</i> (TCa_v3), an early diverging animal that lacks muscle, neurons, and synapses. Co-immunolocalization using antibodies against TCa_v3 and neurosecretory cell marker complexin labeled gland cells, which are hypothesized to play roles in paracrine signaling. Cloning and in vitro expression of TCa_v3 reveals that, despite roughly 600 million years of divergence from other T-type channels, it bears the defining structural and biophysical features of the Ca_v3 family. We also characterize the channel's cation permeation properties and find that its pore is less selective for Ca²⁺ over Na⁺ compared with the human homologue Ca_v3.1, yet it exhibits a similar potent block of inward Na⁺ current by low external Ca²⁺ concentrations (i.e., the Ca²⁺ block effect). A comparison of the permeability features of TCa_v3 with other cloned channels suggests that Ca²⁺ block is a locus of evolutionary change in T-type channel cation permeation properties and that mammalian channels distinguish themselves from invertebrate ones by bearing both stronger Ca²⁺ block and higher Ca²⁺ selectivity. TCa_v3 is the most divergent metazoan T-type calcium channel and thus provides an evolutionary perspective on Ca_v3 channel structure-function properties, ion selectivity, and cellular physiology.