Anatomic locations of the injection sites.

<p>(A) Representative cross section showing the injection sites (fluorescent beads) located within the lateral hypothalamic area (LHA), two injections on each side (note: two injections were overlapping on one side). (B) Schematic corresponding section of A illustrating the injection sites (black) and the distribution of MCH neurons (small grey dots, modified from (Paterson & Hahn 2010)) in the LHA. (C) Representative section and (D) the schematic corresponding section from a rat showing the injection sites localized outside the LHA in the hippocampus. Abbreviations: 3v, third ventricle; f, fornix; hip, hippocampus; mtt, mammillothalamic trac; VMH, ventromedial hypothalamus. Scale bar represents 0.5 mm.</p

The effects of knockdown of pMCH mRNA expression on the expression levels of pMCH, orexin-A and orexin receptor 1 (OX1R) mRNA in the LHA (A) and MCH peptide levels in cerebrospinal fluid (CSF; B).

<p>(Mean ± S.E.M.). The levels of pMCH, orexin-A, and OX1R mRNA expression in the LHA were measured by RT-PCR 48 hrs post injections of aCSF (open bars), pMCH siRNA (filled bars) or scrambled siRNA (hatched bars) into the LHA (A; N = 5 each group; one way ANOVA, Bonferroni’s post hoc test). MCH peptide levels in the CSF were measured by EIA 72 hrs post injections of pMCH siRNA (black bar) or scrambled siRNA (hatched bar) (N = 5 & 6 respectively; unpaired t-test). Significant difference is indicated with *(P<0.05) or **(P<0.01).</p

The effect of knockdown of pMCH mRNA expression on body weight and glucose levels.

<p>Body weight before (filled bars) and 72 hrs after (open bars) pMCH siRNA injection or scrambled siRNA injection (N = 6 each group, paired t-test) (A). Basal glucose levels before (filled bars) and 72 hrs after (open bars) pMCH siRNA or scrambled siRNA injection (N = 6 each group, paired t-test) (B). The pMCH siRNA treated group had significant lower glucose levels than that of scrambled siRNA treated group (B; one way ANONA). (Mean ± S.E.M.) Significant difference is indicated with *(P<0.05).</p

Maintenance or Collapse: Responses of Extraplastidic Membrane Lipid Composition to Desiccation in the Resurrection Plant <i>Paraisometrum mileense</i>

<div><p>Resurrection plants usually grow in specific or extreme habitats and have the capacity to survive almost complete water loss. We characterized the physiological and biochemical responses of <i>Paraisometrum mileense</i> to extreme desiccation and found that it is a resurrection plant. We profiled the changes in lipid molecular species during dehydration and rehydration in <i>P. mileense</i>, and compared these with corresponding changes in the desiccation-sensitive plant <i>Arabidopsis thaliana</i>. One day of desiccation was lethal for <i>A. thaliana</i> but not for <i>P. mileense</i>. After desiccation and subsequent rewatering, <i>A. thaliana</i> showed dramatic lipid degradation accompanied by large increases in levels of phosphatidic acid (PA) and diacylglycerol (DAG). In contrast, desiccation and rewatering of <i>P. mileense</i> significantly decreased the level of monogalactosyldiacylglycerol and increased the unsaturation of membrane lipids, without changing the level of extraplastidic lipids. Lethal desiccation in <i>P. mileense</i> caused massive lipid degradation, whereas the PA content remained at a low level similar to that of fresh leaves. Neither damage nor repair processes, nor increases in PA, occurred during non-lethal desiccation in <i>P. mileense</i>. The activity of phospholipase D, the main source of PA, was much lower in <i>P. mileense</i> than in <i>A. thaliana</i> under control conditions, or after either dehydration or rehydration. It was demonstrated that low rates of phospholipase D-mediated PA formation in <i>P. mileense</i> might limit its ability to degrade lipids to PA, thereby maintaining membrane integrity following desiccation.</p></div

Changes in (A) soluble sugar, (B) proline, (C) malondialdehyde (MDA), and (D) relative electricity conductivity during dehydration (Deh) and rehydration (Reh) in <i>P. mileense</i> and <i>A. thaliana</i> leaves.

<p>Values are means ± standard deviation (<i>n</i>  =  4 or 5). Values in the same bar type with different letters indicate that values are significantly different (<i>P</i> < 0.05). </p

Transphosphatidylation activities of <i>A. thaliana</i> and <i>P. mileense</i>.

<p>Reaction products were separated by thin-layer chromatography and monitored by UV colorimetric analysis.</p

Hierarchical clustering analysis of lipid molecular species during dehydration (Deh) and rehydration (Reh) of <i>P. mileense</i> and <i>A. thaliana</i> leaves.

<p>(A) Contents (nmol/mg dry weight) of lipid molecular species. (B) Compositions (mol %) of lipid molecular species. The colored bar within a column represents the lipid molecular species in the corresponding plants and treatments. The color of each bar represents the abundance of the indicated lipid species, which is expressed relative to the change from the mean center of each lipid species within all treatments. Lipid species in the corresponding lipid classes were sorted using class (as indicated), total acyl carbons (within a class), and total double bonds (with class and total acyl carbons) in ascending order. Values are means (<i>n</i> = 4 or 5).</p

Half-lethal dehydration (Deh 4 d), lethal dehydration (Deh 5 d), and subsequent rehydration (Reh) of <i>P. mileense</i> leaves.

<p>White coloration (upper picture) or low <i>F_v</i>/<i>F_m</i> values for variable fluorescence (lower picture). The color bar at the bottom indicates <i>F_v</i>/<i>F_m</i> values.</p

Amount of lipid in each head-group class and total polar lipid during dehydration (Deh) and rehydration (Reh) of <i>P. mileense</i> and <i>A. thaliana</i> leaves.

<p>The percentage relative change in lipids of dehydration RC (F–D) is the value for the difference between the values of Fresh and Deh discs, divided by the value of Fresh discs; that of rehydration RC (D–R) is the value for the significant difference between the values of Deh and Reh discs, divided by the value of Deh discs. Values in the same row with different letters are significantly different (<i>P</i><0.05). Values are means ± standard deviation (<i>n</i> = 4 or 5).</p

Changes of pigments during dehydration (Deh) and rehydration (Reh) in <i>P. mileense</i> and <i>A. thaliana</i> leaves.

<p>Values in the same row with different letters are significantly different (<i>P</i><0.05). Values are means ± SD (<i>n</i> = 4 or 5).</p