Hv1/VsoP is a dimeric voltage-gated H + channel in which the gating of one subunit is reportedly coupled to that of the other subunit within the dimer. The molecular basis for dimer formation and intersubunit coupling, however, remains unknown. Here we show that the carboxy terminus ends downstream of the s4 voltage-sensor helix twist in a dimer coiled-coil architecture, which mediates cooperative gating. We also show that the temperature-dependent activation of H + current through Hv1/VsoP is regulated by thermostability of the coiled-coil domain, and that this regulation is altered by mutation of the linker between s4 and the coiled-coil. Cooperative gating within the dimer is also dependent on the linker structure, which circular dichroism spectrum analysis suggests is α-helical. our results indicate that the cytoplasmic coiled-coil strands form continuous α-helices with s4 and mediate cooperative gating to adjust the range of temperatures over which Hv1/VsoP operates.

The mammalian habenula consists of the medial and lateral habenulae. Recent behavioral and electrophysiological studies suggested that the lateral habenula plays a pivotal role in controlling motor and cognitive behaviors by influencing the activity of dopaminergic and serotonergic neurons. Despite the functional significance, manipulating neural activity in this pathway remains difficult because of the absence of a genetically accessible animal model such as zebrafish. To address the level of lateral habenula conservation in zebrafish, we applied the tract-tracing technique to GFP (green fluorescent protein)-expressing transgenic zebrafish to identify habenular neurons that project to the raphe nuclei, a major target of the mammalian lateral habenula. Axonal tracing in live and fixed fish showed projection of zebrafish ventral habenula axons to the ventral part of the median raphe, but not to the interpeduncular nucleus where the dorsal habenula projected. The ventral habenula expressed protocadherin 10a, a specific marker of the rat lateral habenula, whereas the dorsal habenula showed no such expression. Gene expression analyses revealed that the ventromedially positioned ventral habenula in the adult originated from the region of primordium lateral to the dorsal habenula during development. This suggested that zebrafish habenulae emerge during development with mediolateral orientation similar to that of the mammalian medial and lateral habenulae. These findings indicated that the lateral habenular pathways are evolutionarily conserved pathways and might control adaptive behaviors in vertebrates through the regulation of monoaminergic activities.

The mammalian habenula is involved in regulating the activities of serotonergic and dopaminergic neurons. It consists of the medial and lateral habenulae, with each subregion having distinct neural connectivity. Despite the functional significance, manipulating neural activity in a subset of habenular pathways remains difficult because of the poor availability of molecular markers that delineate the subnuclear structures. Thus, we examined the molecular nature of neurons in the habenular subnuclei by analyzing the gene expressions of neurotransmitter markers. The results showed that different subregions of the medial habenula (MHb) use different combinations of neurotransmitter systems and could be categorized as either exclusively glutamatergic (superior part of MHb), both substance P-ergic and glutamatergic (dorsal region of central part of MHb), or both cholinergic and glutamatergic (inferior part, ventral region of central part, and lateral part of MHb). The superior part of the MHb strongly expressed interleukin-18 and was innervated by noradrenergic fibers. In contrast, the inferior part, ventral region of the central part, and lateral part of the MHb were peculiar in that acetylcholine and glutamate were cotransmitted from the axonal terminals. In contrast, neurons in the lateral habenula (LHb) were almost uniformly glutamatergic. Finally, the expressions of Htr2c and Drd2 seemed complementary in the medial LHb division, whereas they coincided in the lateral division, suggesting that the medial and lateral divisions of LHb show strong heterogeneity with respect to monoamine receptor expression. These analyses clarify molecular differences between subnuclei in the mammalian habenula that support their respective functional implications.

The mammalian habenula is involved in regulating the activities of serotonergic and dopaminergic neurons. It consists of the medial and lateral habenulae, with each subregion having distinct neural connectivity. Despite the functional significance, manipulating neural activity in a subset of habenular pathways remains difficult because of the poor availability of molecular markers that delineate the subnuclear structures. Thus, we examined the molecular nature of neurons in the habenular subnuclei by analyzing the gene expressions of neurotransmitter markers. The results showed that different subregions of the medial habenula (MHb) use different combinations of neurotransmitter systems and could be categorized as either exclusively glutamatergic (superior part of MHb), both substance P‐ergic and glutamatergic (dorsal region of central part of MHb), or both cholinergic and glutamatergic (inferior part, ventral region of central part, and lateral part of MHb). The superior part of the MHb strongly expressed interleukin‐18 and was innervated by noradrenergic fibers. In contrast, the inferior part, ventral region of the central part, and lateral part of the MHb were peculiar in that acetylcholine and glutamate were cotransmitted from the axonal terminals. In contrast, neurons in the lateral habenula (LHb) were almost uniformly glutamatergic. Finally, the expressions of Htr2c and Drd2 seemed complementary in the medial LHb division, whereas they coincided in the lateral division, suggesting that the medial and lateral divisions of LHb show strong heterogeneity with respect to monoamine receptor expression. These analyses clarify molecular differences between subnuclei in the mammalian habenula that support their respective functional implications. J. Comp. Neurol. 520:4051–4066, 2012. © 2012 Wiley Periodicals, Inc.

(13)C nuclear magnetic resonance measurements were performed on κ-(BEDT-TTF)(2)Cu(NCS)(2), with the external field placed parallel to the quasi-2D conducting layers. The absorption spectrum is used to determine the electronic spin polarization M(s) as a function of external field H at a temperature T=0.35 K. A discontinuity in the derivative dM(s)/dH at an applied field of H(s)=213±3 kOe is taken as evidence for a Zeeman-driven transition within the superconducting state and stabilization of inhomogeneous superconductivity.