Immunofluorescent staining of cultured NG neurons.

<p>NG neurons were stained using antibodies against P2X1, P2X2, P2X3, and P2X4 receptor subunits (A, D, G, and J, respectively). The nuclei of cultured NG neurons were stained with antibodies against NeuN (B, E, H, K). Merged images (C, F, I, L) representing co-staining of P2X receptor subunits and NeuN are shown. The scale bar shown in L is representative of all images, and represents 50 µm.</p

Four types of I_ATPs and their relationship with NG neuron diameter.

<p>(A) The currents activated by ATP (10⁻⁴ M) in NG were characterized according to fast (F), intermediate (I), slow (S), and very slow (VS) kinetics with an expanded time axis. The horizontal bar above the traces indicates the application of ATP. (B) The 10–90% rising time (ordinate) of different types of ATP (10⁻⁴ M)-activated currents (black circles  =  type F, red triangles  =  type I, green squares  =  for type S, and yellow diamonds  =  type VS I_ATPs) against neuron diameter. Each point in the graph represents a single cell. The correlation coefficient was 0.66 (<i>p</i><0.05). (C) Representative traces of type F, I, S, and VS ATP (10⁻⁴ M)-activated currents. (D) Comparisons of the absolute ratios of the four types of I_ATPs in different subpopulations (small, medium, and large sizes) of NG neurons. Specifically, F type: small (13/54, 28.9%), medium (36/54, 80.0%), and large (5/45, 11.1%); I type: small (5/75, 6.7%), medium (56/75, 74.7%), and large (14/75, 18.6%); S type: small (11/126, 8.7%), medium (94/126, 74.6%), and large (21/126, 16.7%); VS type: small (7/59, 11.9%), medium (34/59, 57.6%), and large (18/59, 30.5%).</p

The frequency distribution of the area-size of P2X1–4 subunit-positive neurons.

<p>(A), (B), (C), and (D) show P2X–IR sections that were counterstained with neutral red to precisely calculate the number of P2X1, P2X2, P2X3, and P2X4-positive neurons, respectively (using black positive cells compared with red negative cells). Scale bar in (D)  = 100 µm. (E), (F), (G), and (H) represent the frequency distribution of the area-size of P2X1, P2X2, P2X3, and P2X4-IR, respectively.</p

Concentration-response relationships and the efficacy order of P2X receptor antagonists on I_ATPs.

<p>(A) Sequential current traces of F, I, S, and VS ATP-activated currents recorded from rat NG neurons in response to different concentrations of ATP (from 10⁻⁵ to 3×10⁻³ M). Current traces of each type were obtained from the same neuron. (B) The dose-response curves for each type of I_ATPs. Each point represents the means ± SEM of 10–15 neurons. All ATP-induced currents were normalized to the response induced by 3×10⁻³ M ATP in each type. The holding potential was set at −60 mV. The data for ATP were a good fit to the Hill equation I = I_max/[1+ (EC50/C) n], where C is the concentration of ATP, I is the normalized amplitude of I_ATP, and EC50 is the concentration of ATP for the half maximal current response. (C) The efficacy order of the inhibitory effects of P2X receptor antagonists on four distinct I_ATPs. The columns in the bar graph show the inhibitory effects of the P2X receptor antagonists: PPADS (10⁻⁴M), suramin (10⁻⁴ M), and RB2 (10⁻⁴ M). F-type, suramin >PPADS > RB2; I-type, suramin > PPADS > RB2; S-type, suramin > RB2> PPADS; VS-type: suramin > RB2> PPADS. *<i>p</i><0.05, **<i>p</i><0.01.</p

Distribution and expression of P2X1–4 subunits in NG tissue.

<p>(A) A schematic diagram of the maximum cross section of a rat nodose ganglion (A, corresponding to panel B). (B) Representative graph of the distribution of P2X receptor-positive cells throughout the whole nodose ganglion section under a 20× light microscopic field. (C–F) Immunohistochemical staining using polyclonal antibodies against P2X1 (C), P2X2 (D), P2X3 (E), and P2X4 (F); 100× magnification. Scale bars in B = 500 µm and F = 100 µm.</p

Relationship between the numbers and sizes of four types of I_ATP NG neurons.

<p>Relationship between the numbers and sizes of four types of I_ATP NG neurons.</p

The proportion of P2X1–4 subunit-expressing neurons compared with the total number of NG neurons.

<p>The upper panel shows immunohistochemical staining for P2X1–4 subunits in the maximum cross section of the NG tissues. The lower panel shows that P2X3-IR was the most prevalent neuronal subtype, following by P2X2-IR; P2X4-IR had the lowest prevalence. *<i>p</i><0.05.</p

Relevance of the P2X1–4 subunits on the four types of I_ATP.

<p>(A) Schematic view of the setup for the whole cell patch clamp and a representative image of a recorded cell under the phase contrast microscope and immunohistochemistry. (B) Immunohistochemistry revealed positive or negative staining for P2X1–4 subunits, which correlated with the type of I_ATP and cell size. The samples in each row were from four different neurons that responded to ATP with different types of ATP-activated current. P2X3 staining was positive in all four types of I_ATP neurons. P2X1 was positive in F, I, and S I_ATPs, but negative in VS. P2X2 staining was only absent in neurons with type I I_ATP, and P2X4 was positive in neurons with type F, I, and some S I_ATPs.</p

Cyclic Phase Transition from Hexagonal to Orthorhombic Then Back to Hexagonal of EuF₃ While Loading Uniaxial Pressure and under High Temperature

The structure and photoluminescence properties are investigated under high pressure and high temperature for pure orthorhombic and hexagonal EuF₃ nanocrystals. Under hydrostatic compression, the hexagonal EuF₃ remains stable at pressures up to 26 GPa. Under nonhydrostatic compression, a cyclic phase transition from hexagonal to orthorhombic and then back to hexagonal is observed for the first time. When loading uniaxial compression, the pure hexagonal EuF₃ partly transforms to orthorhombic at 70 MPa, then the orthorhombic EuF₃ transforms to hexagonal at about 3 GPa, and the transition is completed at about 10 GPa. The cyclic phase transition is also observed during the heating process; the hexagonal transforms to orthorhombic at 550 °C and then to hexagonal at 855 °C. The content phase diagrams are obtained under high pressure and at high temperature

Guiding the Formation of Metal–Organic Structures of 1,4-Diaminoanthraquinone through Surface-Based Cu Atoms

The gradual guidance of the formation of metal–organic structures through surface-based Cu atoms for 1,4-diaminoanthraquinones (DAQs) has been studied by scanning tunneling microscopy (STM) at room temperature. On the Ag(110) surface, the transition from a hydrogen-bond network structure to metal–organic coordination structures of DAQs can be induced by introducing foreign copper atoms. Due to the weak interaction between DAQs and Ag(110), thermal treatment easily leads to the desorption of DAQs from the surface. To address this challenge, Cu(111) is selected as the substrate. Under thermal driving and in the presence of copper adatoms, the hydrogen-bond network structure of DAQs on the surface gradually undergoes a transition into a metal-coordinated structure, eventually leading to the formation of metal–organic complexes through amino dehydrogenation. It is demonstrated that the construction of a metal–organic coordination structure on metal surfaces is a result of the competition among factors such as metal atoms, functional groups of molecules, surface chemical activity, and temperature