Receptive field size reduction in ON- and OFF-RGCs.

<p>The abscissa is animal age (wk) and vertical coordinate depicts relative percentage of RF size reduction. Filled diamond and solid line represents ON-RGC while open circle and dashed line shows OFF-RGC RF size reduction.</p

Visual response properties of ON- and OFF-RGCs of 20 wk db/db mice after in vitro SOD application.

<p><b>Panel A:</b> Discharge patterns of two ON-center RGCs in response to 0.02 cyc/deg sinusoidal drifting gratings of different contrast levels. The top two traces exhibit discharge patterns of an ON-RGC (305-C3, top left) recorded from a db/m retina (eccentricity: 0.94 mm) and bottom two traces show discharge patterns of an ON-RGC (121-C3, bottom left) recorded from a db/db retina (eccentricity: 1.17 mm). Contrast levels are shown to the left of the response traces. <b>Panel B:</b> Discharge patterns of two OFF-RGCs. The top two traces exhibit discharge patterns of an OFF-RGC (307-C3, top right) recorded from a db/m retina (eccentricity: 1.71 mm) and bottom two traces show discharge patterns of an OFF-RGC (121-C4) recorded from a db/db retina (eccentricity: 1.21 mm). <b>Panel C:</b> contrast response profiles of the two ON cells exhibited in Panel A. The abscissa is contrast (%) and vertical coordinate depicts contrast response magnitude (spk/sec). The open triangles depict db/m ON; open squares represent db/db ON. <b>Panel D:</b> The contrast response profiles of two OFF-RGCs illustrated in Panel B. filled triangles: db/m OFF; and filled squares: db/db OFF. <b>Panel E:</b> Histogram comparing average contrast sensitivity of ON and OFF RGCs of within each group. For db/db mice, as a comparison was made between the control (without SOD treatment) and ON-RGCs that received SOD treatment, the average contrast gain of latter was significantly increased (p<0.05, n = 6). As a comparison was made between the control and OFF-RGCs that received SOD treatment, the average contrast sensitivity of latter was significantly improved (p<0.01, n = 6). For db/m mice, after the SOD treatment, the contrast gain of ON-RGCs was not significantly changed whereas that of OFF-RGCs was significantly suppressed (p<0.01, n = 7). <b>Panel F:</b> Histogram comparing the average RF size of RGCs after SOD treatment. Other conventions are as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030343#pone-0030343-g004" target="_blank">Figure 4</a>.</p

Contrast responses of RGCs recorded from 20 wk old db/m and db/db retinas.

<p><b>Panel A:</b> Discharge patterns of two ON-center RGCs in response to 0.02 cyc/deg sinusoidal drifting gratings at two different contrast levels, 60% and 20%, respectively. The top two traces exhibit discharge patterns of an ON-RGC (229-C1, top left) recorded from a db/m retina (eccentricity: 1.27 mm) and bottom two traces show discharge patterns of an ON-RGC (129-C2, bottom left) recorded from a db/db retina (eccentricity: 1.05 mm). Contrasts are shown to the left of the response traces. <b>Panel B:</b> Discharge patterns of two OFF-RGCs. The top two traces exhibit discharge patterns of an OFF-RGC (408-C6, top right) recorded from a db/m retina (eccentricity: 0.95 mm) and the bottom two traces show discharge patterns of an OFF-RGC (024-C2) recorded from a db/db retina (eccentricity: 1.41 mm). <b>Panel C:</b> contrast gain of the two ON cells exhibited in Panel A. The abscissa is the contrast and vertical coordinate depicts contrast response magnitude (spk/sec). The open triangles depict db/m ON; open squares represent db/db ON. <b>Panel D:</b> compares the contrast gain of two OFF-RGCs illustrated in Panel B: filled triangles: db/m OFF; and filled squares: db/db OFF. Panel E: Histogram comparing average contrast gains of ON and OFF RGCs of within each group. Other conventions are as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030343#pone-0030343-g002" target="_blank">Figure 2</a>.</p

ROS levels of retinal neurons before and after SOD treatment in 8 and 20 wk db/m and db/db mice.

<p>In situ ROS generation was determined in freshly prepared retinal sections by immunohistochemical staining for dihydroethidium (DHE). The quantitatively analyze was performed by using an image analysis software (Adobe Photoshop CS5, CA). The thickness of each retinal section was 10 µm. Micrographs of retinal sections that were stained with DHE: <b>Panel A:</b> a: 8 wk db/m; b: 20 wk db/m; c: 20 wk db/m, 2-hour SOD treatment; d: 20 wk db/m 5-hour SOD treatment; e: 8 wk db/db; f: 20 wk db/db; g: 20 wk db/db, 2-hour SOD treatment; h: 20 wk db/db, 5-hour SOD treatment. ONL: outer nucleus layer; OPL: outer plexiform layer; INL: inner nucleus layer; GCL: ganglion cell layer. <b>Panel B:</b> quantitative analysis of retinal ROS levels. The fluorescent intensity of DHE labeled neurons was quantified (means±SEM, n = 4). In comparison with db/m mice, the intracellular ROS generation was significantly increased in retinas of 8 and 20 wk db/db mice (8 wk: *p<0.05; 20 wk: ***p<0.0001, one-way ANOVA). <b>Panel C:</b> SOD treatment for 2- and 5- hour significantly reduced the ROS levels in 20 wk db/db mice retina (2 hours: **p<0.001; 5 hours: ***p<0.0001, one-way ANOVA). Scale bar: 20 µm. Other conventions are as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030343#pone-0030343-g002" target="_blank">Figures 2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030343#pone-0030343-g003" target="_blank">3</a>.</p

Average receptive field diameter of retinal ganglion cells in db/m and db/db mice.

<p>Values are means±SEM.</p><p>**P<0.001</p><p>*p<0.05 vs dm.</p

Animal body weight and glucose level.

<p>Values are means±SEM.</p><p>***p<0.0001</p><p>*p<0.05 vs dm.</p

image_3.jpg

<p>Evidence is accumulating that group 2 innate lymphoid cells (ILC2) play an important role in allergic airway inflammation by producing a large amount of type 2 cytokines. But it remains poorly understood how its activities are properly controlled in vivo. Here, we demonstrated that prostaglandin E₂ (PGE₂) had a profound inhibitory effect on IL-33-induced ILC2 expansion and IL-5 and IL-13 production in vitro. This effect was mimicked by PGE₁-alcohol but attenuated by ONO-AE3-208, indicating a selective action through the E-prostanoid 4 (EP4) receptor. In the IL-33-induced asthma model, coadministration of PGE₂ or PGE₁-alcohol resulted in diminished IL-5 and IL-13 production, reduced eosinophilia and alleviated lung pathology. In contrast, EP4-deficient mice displayed an exacerbated inflammatory response in another ILC2-mediated asthma model induced by Alternaria extract. Mechanistic studies demonstrated that the PGE₂-mediated inhibition of ILC2 was dependent on cyclic adenosine monophosphate (cAMP) production. Further downstream, PGE₂-EP4-cAMP signaling led to suppression of GATA3 and ST2 expression, which is known to be critical for ILC2 activation. These findings reveal a novel function of PGE₂ as a negative regulator of ILC2 activation and highlight an endogenous counter-regulatory mechanism for the control of innate allergic inflammatory responses.</p

image_2.jpg

<p>Evidence is accumulating that group 2 innate lymphoid cells (ILC2) play an important role in allergic airway inflammation by producing a large amount of type 2 cytokines. But it remains poorly understood how its activities are properly controlled in vivo. Here, we demonstrated that prostaglandin E₂ (PGE₂) had a profound inhibitory effect on IL-33-induced ILC2 expansion and IL-5 and IL-13 production in vitro. This effect was mimicked by PGE₁-alcohol but attenuated by ONO-AE3-208, indicating a selective action through the E-prostanoid 4 (EP4) receptor. In the IL-33-induced asthma model, coadministration of PGE₂ or PGE₁-alcohol resulted in diminished IL-5 and IL-13 production, reduced eosinophilia and alleviated lung pathology. In contrast, EP4-deficient mice displayed an exacerbated inflammatory response in another ILC2-mediated asthma model induced by Alternaria extract. Mechanistic studies demonstrated that the PGE₂-mediated inhibition of ILC2 was dependent on cyclic adenosine monophosphate (cAMP) production. Further downstream, PGE₂-EP4-cAMP signaling led to suppression of GATA3 and ST2 expression, which is known to be critical for ILC2 activation. These findings reveal a novel function of PGE₂ as a negative regulator of ILC2 activation and highlight an endogenous counter-regulatory mechanism for the control of innate allergic inflammatory responses.</p

image_4.jpg

<p>Evidence is accumulating that group 2 innate lymphoid cells (ILC2) play an important role in allergic airway inflammation by producing a large amount of type 2 cytokines. But it remains poorly understood how its activities are properly controlled in vivo. Here, we demonstrated that prostaglandin E₂ (PGE₂) had a profound inhibitory effect on IL-33-induced ILC2 expansion and IL-5 and IL-13 production in vitro. This effect was mimicked by PGE₁-alcohol but attenuated by ONO-AE3-208, indicating a selective action through the E-prostanoid 4 (EP4) receptor. In the IL-33-induced asthma model, coadministration of PGE₂ or PGE₁-alcohol resulted in diminished IL-5 and IL-13 production, reduced eosinophilia and alleviated lung pathology. In contrast, EP4-deficient mice displayed an exacerbated inflammatory response in another ILC2-mediated asthma model induced by Alternaria extract. Mechanistic studies demonstrated that the PGE₂-mediated inhibition of ILC2 was dependent on cyclic adenosine monophosphate (cAMP) production. Further downstream, PGE₂-EP4-cAMP signaling led to suppression of GATA3 and ST2 expression, which is known to be critical for ILC2 activation. These findings reveal a novel function of PGE₂ as a negative regulator of ILC2 activation and highlight an endogenous counter-regulatory mechanism for the control of innate allergic inflammatory responses.</p

Silencing of Long Noncoding RNA AK139328 Attenuates Ischemia/Reperfusion Injury in Mouse Livers

<div><p>Recently, increasing evidences had suggested that long noncoding RNAs (LncRNAs) are involved in a wide range of physiological and pathophysiological processes. Here we determined the LncRNA expression profile using microarray technology in mouse livers after ischemia/reperfusion treatment. Seventy one LncRNAs were upregulated, and 27 LncRNAs were downregulated in ischemia/reperfusion-treated mouse livers. Eleven of the most significantly deregulated LncRNAs were further validated by quantitative PCR assays. Among the upregulated LncRNAs confirmed by quantitative PCR assays, AK139328 exhibited the highest expression level in normal mouse livers. siRNA-mediated knockdown of hepatic AK139328 decreased plasma aminotransferase activities, and reduced necrosis area in the livers with a decrease in caspase-3 activation after ischemia/reperfusion treatment. In ischemia/reperfusion liver, knockdown of AK139328 increased survival signaling proteins including phosphorylated Akt (pAkt), glycogen synthase kinase 3 (pGSK3) and endothelial nitric oxide synthase (peNOS). Furthermore, knockdown of AK139328 also reduced macrophage infitration and inhibited NF-κB activity and inflammatory cytokines expression. In conclusion, these findings revealed that deregulated LncRNAs are involved in liver ischemia/reperfusion injury. Silencing of AK139328 ameliorated ischemia/reperfusion injury in the liver with the activation of Akt signaling pathway and inhibition of NF-κB activity. LncRNA AK139328 might be a novel target for diagnosis and treatment of liver surgery or transplantation. </p> </div