Region- and neuronal-subtype-specific expression of Na,K-ATPase alpha and beta subunit isoforms in the mouse brain.

Na,K-ATPase is a ubiquitous molecule contributing to the asymmetrical distribution of Na+ and K+ ions across the plasma membrane and maintenance of the membrane potential, a prerequisite of neuronal activity. Na,K-ATPase comprises three subunits (α, ß, and FXYD). The α subunit has four isoforms in mice, with three of them (α1, α2, and α3) expressed in the brain. However, the functional and biological significances of the different brain isoforms remain to be fully elucidated. Recent studies have revealed the association of Atp1a3, a gene encoding α3 subunit, with neurological disorders. To map the cellular distributions of the α subunit isoforms and their coexpression patterns, we evaluated the mRNA expression of Atp1a1, Atp1a2, and Atp1a3 by in situ hybridization in the mouse brain. Atp1a1 and Atp1a3 were expressed in neurons, whereas Atp1a2 was almost exclusively expressed in glial cells. Most neurons coexpressed Atp1a1 and Atp1a3, with highly heterogeneous expression levels across the brain regions and neuronal subtypes. We identified parvalbumin (PV)-expressing GABAergic neurons in the hippocampus, somatosensory cortex, and retrosplenial cortex as an example of a neuronal subtype expressing low Atp1a1 and high Atp1a3. The expression of Atp1b isoforms was also heterogeneous across brain regions and cellular subtypes. The PV-expressing neurons expressed a high level of Atp1b1 and a low level of Atp1b2 and Atp1b3. These findings provide basic information on the region- and neuronal-subtype-dependent expression of Na,K-ATPase α and ß subunit isoforms, as well as a rationale for the selective involvement of neurons expressing high levels of Atp1a3 in neurological disorders.

GABAergic neurons in the olfactory cortex projecting to the lateral hypothalamus in mice.

Olfaction guides goal-directed behaviours including feeding. To investigate how central olfactory neural circuits control feeding behaviour in mice, we performed retrograde tracing from the lateral hypothalamus (LH), an important feeding centre. We observed a cluster of retrogradely labelled cells distributed in the posteroventral region of the olfactory peduncle. Histochemical analyses revealed that the majority of these retrogradely labelled projection neurons expressed glutamic acid decarboxylase 65/67 (GAD65/67), but not vesicular glutamate transporter 1 (VGluT1). We named this region containing GABAergic projection neurons the ventral olfactory nucleus (VON) to differentiate it from the conventional olfactory peduncle. VON neurons were less immunoreactive for DARPP-32, a striatal neuron marker, compared to neurons in the olfactory tubercle and nucleus accumbens, which distinguished the VON from the ventral striatum. Fluorescent labelling confirmed putative synaptic contacts between VON neurons and olfactory bulb projection neurons. Rabies-virus-mediated trans-synaptic labelling revealed that VON neurons received synaptic inputs from the olfactory bulb, other olfactory cortices, horizontal limb of the diagonal band, and prefrontal cortex. Collectively, these results identify novel GABAergic projection neurons in the olfactory cortex that may integrate olfactory sensory and top-down inputs and send inhibitory output to the LH, which may modulate odour-guided LH-related behaviours.

Opposing Roles of Dopamine Receptor D1- and D2-Expressing Neurons in the Anteromedial Olfactory Tubercle in Acquisition of Place Preference in Mice.

Olfaction induces adaptive motivated behaviors. Odors associated with food induce attractive behavior, whereas those associated with dangers induce aversive behavior. We previously reported that learned odor-induced attractive and aversive behaviors accompany activation of the olfactory tubercle (OT) in a domain- and cell type-specific manner. Odor cues associated with a sugar reward induced attractive behavior and c-fos expression in the dopamine receptor D1-expressing neurons (D1 neurons) in the anteromedial OT. In contrast, odor cues associated with electrical shock induced aversive behavior and c-fos expression in the pamine receptor D2-expressing neurons (D2 neurons) in the anteromedial OT, as well as the D1 neurons in the lateral OT. Here, we investigated whether the D1 and D2 neurons in the anteromedial OT play distinct roles in attractive or aversive behaviors, using optogenetic stimulation and real-time place preference (RTPP) tests. Mice expressing ChETA (ChR2/E123T)-enhanced yellow fluorescent protein (EYFP) in the D1 neurons in the anteromedial OT spent a longer time in the photo-stimulation side of the place preference chamber than the control mice expressing EYFP. On the other hand, upon optogenetic stimulation of the D2 neurons in the anteromedial OT, the mice spent a shorter time in the photo-stimulation side than the control mice. Local neural activation in the anteromedial OT during the RTPP tests was confirmed by c-fos mRNA expression. These results suggest that the D1 and D2 neurons in the anteromedial OT play opposing roles in attractive and aversive behaviors, respectively.

Fish oil feeding reverses hepatomegaly and disrupted hepatic function due to the lack of FXR signaling.

Mice lacking farnesoid X receptor (FXR) are used as a model for nonalcoholic fatty liver disease because their livers exhibit hepatomegaly, hepatic steatosis, and hepatic inflammation. The influence of fish oil feeding on hepatomegaly and disrupted hepatic function was investigated using female Fxr-null mice and wild-type mice fed a fish oil diet (2% fish oil and 2% corn oil) or a control diet (4% corn oil) for 4 weeks. Hepatic n-3 polyunsaturated fatty acid (PUFA) levels, including 22:6 n-3 docosahexaenoic acid (DHA) and 20:5 n-3 eicosapentaenoic acid (EPA) were significantly higher in the fish oil group than those in the control group of Fxr-null mice and wild-type mice. Fxr-null mouse livers of the control group showed a whitish brown coloration, whereas Fxr-null mouse livers of the fish oil group showed a dark brown coloration similar to that of wild-type mice. The liver in Fxr-null mice of the fish oil group was smaller than that of the control group. There was a significant decrease in the levels of hepatic damage-associated diagnostic markers, hepatic and serum bile acids, triglycerides, free fatty acids, and total cholesterol levels in Fxr-null mice because of fish oil feeding. It also reversed elevated mRNA levels of oxidative stress-related genes (Hmox1, Gsta1, and Gsta2) and reduced mRNA levels of transcriptional factors (Pparα and Shp) in Fxr-null mice. These results suggest that fish oil feeding reverses hepatomegaly and disrupted hepatic function due to the lack of FXR signaling.

Large, negative magnetoresistance in an oleic acid-coated Fe(3)O(4) nanocrystal self-assembled film.

An oleic acid-coated Fe3O4 nanocrystal self-assembled film was fabricated via drop casting of colloidal particles on a SiO2/Si substrate. The film exhibited bifurcation of the zero-field-cooled and field-cooled magnetizations around 250 K. The nonlinear current-voltage (I-V) characteristics between the source and drain electrodes in both zero and non-zero magnetic fields (H) were observed above and below the bifurcation temperature. A large negative magnetoresistance (MR ≈ -60%) was achieved at 200 K and H = 1 T. Even at 295 K and 0.2 T, the negative MR (∼ -50%) persisted. A Fowler-Nordheim plot and power-law scaling of the I-V characteristics revealed that the current flows through two-dimensional (2D) percolated electron tunneling paths. The enlargement of MR can be attributed to spin-dependent electron tunneling between magnetically coupled Fe3O4 nanocrystals self-assembled in 2D ordered arrays.

Magnetic and magnetoelectric properties of self-assembled Fe2.5Mn0.5O4 nanocrystals.

We report magnetoresistance of -40%, corresponding to 80% spin polarization, at magnetic field of 0.5 T and 200 K for oleic acid-coated Fe(2.5)Mn(0.5)O(4) nanocrystals (FMO NCs) self-assembled on a SiO(2)/Si substrate by drop casting fabrication. The FMO NCs exhibited spin glass transition around 150 K and nonlinear current-voltage (I-V) characteristics. Fowler-Nordheim plot of the I-V characteristics indicated that electrons tunnel directly barriers between the FMO NCs. Transmission electron microscopy revealed that the FMO NCs are elongated hexagon in shape with size of ∼15 × 20 nm. The FMO NCs self-assembled in two-dimension hexagonal networks of collinear ferromagnetic moments. The [111] easy magnetization axis of each FMO NC was parallel to each other in the hexagonal arrays. Geometrically frustrated lattice of collinear ferromagnetic moments supports both a low and a high intergranular tunneling conductance for the self-assembled FMO NCs without and with magnetic fields, respectively.