Phylogenetic position of <i>M</i>. <i>desjardinsi Spiroplasma</i> among <i>Spiroplasma</i> bacteria and relatives based on spot gene sequences.

<p>A maximum likelihood tree (lnL = −2525.6794) is shown. Bootstrap probabilities are given at the nodes. <i>M</i>. <i>desjardinsi Spiroplasma</i> is depicted as red. The scale bar indicates the number of nucleotide substitutions per site. A clade containing <i>S</i>. <i>poulsonii</i>, <i>S</i>. <i>insolitum</i>, <i>S</i>. <i>kunkelii</i>, <i>S</i>. <i>phoeniceum</i>, <i>S</i>. <i>citri</i> and <i>S</i>. <i>melliferum</i> is shaded with light blue. A host organism is given in parenthesis. An accession number is given at the end of each OTU. MK represents a male-killer.</p

Survivorship of offspring produced by wild-caught females.

<p>Proportion of surviving individuals at the first-instar larval stage (II), the pupal stage (III), the pharate-adult (i.e., moving pupal) stage (IV), and the adult stage (V) among embryos (I). A solid line (red) represents a brood that developed to only females. A broken line (blue) represents a brood that developed to males and females with a ratio not significantly deviated from 1:1 (<i>P</i> > 0.05). A photograph of an adult female is given in the inset.</p

qPCR-estimates of endosymbiont population densities in mothers that are categorized by their offspring sex ratio.

<p>(a) <i>Spiroplasma</i> density in mothers (i.e., Relative values of <i>Spiroplasma spoT</i> gene copies per mitochondrial COII gene copy). (b) <i>Rickettsia</i> density in mothers (i.e., Relative values of <i>Rickettsia gltA</i> gene copies per mitochondrial COII gene copy).</p

Effects of tetracycline on survival rates and sex ratio of the matriline MK20.

<p>Effects of tetracycline on survival rates and sex ratio of the matriline MK20.</p

Transmission electron micrographs of <i>Spiroplasma</i> and <i>Rickettsia</i> in the female reproductive tissues of <i>M</i>. <i>desjardinsi</i>.

<p><b>a</b>: Ovaries of a female <i>M</i>. <i>desjardinsi</i>. A circle represents a portion subjected to electron microscopy. <b>b</b>: Undifferentiated part of an ovariole, with <i>Rickettsia</i>-like structure in the follicle cells and <i>Spiroplasma</i>-like structure in the oocyte. <b>c</b>,<b>d, e</b>: Magnified images of (b), showing <i>Spiroplasma</i>-like structure. <b>f</b>: Differentiating part of an ovariole, with <i>Rickettsia</i>-like structure in the oocyte and <i>Spiroplasma</i>-like structure in the sheath. S: <i>Spiroplasma</i>; R: <i>Rickettsia</i>; oc: oocyte; fc: follicle cell; sh: sheath; n: nucleus.</p

Infection status of wild-caught <i>M</i>. <i>desjardinsi</i> females and the sex ratio of their offspring.

<p>Infection status of wild-caught <i>M</i>. <i>desjardinsi</i> females and the sex ratio of their offspring.</p

Further images of <i>Spiroplasma</i> and <i>Rickettsia</i> in the female reproductive tissues of <i>M</i>. <i>desjardinsi</i> observed by transmission electron microscopy.

<p><b>a</b>: Differentiating part of an ovariole, with <i>Spiroplasma</i>-like structure outside and inside the sheath and <i>Rickettsia</i>-like structure in the follicle cells. <b>b</b>: Magnified image of (a), showing the presence of <i>Spiroplasma</i>-like structure in the intercellular space and the sheath, possibly at the timing of intrusion into a follicle cell. <b>c</b>: Cytoplasm of an oocyte, showing <i>Rickettsia</i> and <i>Spiroplasma</i>-like structure. <b>d</b>: <i>Spiroplasm</i>-like structure possibly at the timing of intrusion into the oocyte. S: <i>Spiroplasma</i>; R: <i>Rickettsia</i>; oc: oocyte; fc: follicle cell; sh: sheath; n: nucleus.</p

Intraplantar carrageenan injection induces inflammation-mediated mechanical hyperalgesia.

<p>Paw withdrawal thresholds (PWT) following saline or carrageenan injection were determined using the von Frey test on the ipsilateral (A) and contralateral sides (B). (n = 25 mice for saline and n = 27 mice for carrageenan). *p < 0.05 compared with the respective saline-treated mice (two-way ANOVA with the Bonferroni post hoc test).</p

Cav3.2 protein expression is enhanced in ipsilateral L5 DRG tissues during the sub-acute phase (Day 2) following carrageenan injection.

<p>Cav3.2 protein expression was analyzed and normalized relative to the expression of the housekeeping protein ß-actin. The inset of a representative, original immunoblot shows the up-regulation of Cav3.2 protein (n = 6 mice for each group). *p < 0.05 compared with the saline control (unpaired Student's <i>t-test</i>).</p

Cav3.2 mRNA is expressed in a subset of DRG neurons in normal mice.

<p>(A and B) Macroautoradiographic images of an [³⁵S]-labeled probe from an <i>in situ</i> hybridization showing the specificity of the Cav3.2 probe (A; sense probe) and Cav3.2 mRNA expression (B; anti-sense probe) in mouse DRG tissues. Hybridization signals were obtained only when the anti-sense probe was used. The medium DRG neurons (arrows) were densely labeled, whereas smaller neurons (arrow head) were only weakly labeled. Scale bar = 50 μm (C) Microscopic images of a DIG-labeled probe for <i>in situ</i> hybridization (n = 3 mice for each probe). Scale bar = 50 μm. (D) The proportion of Cav3.2 mRNA-positive neurons relative to the total number of DRG neurons was determined. (E) Histogram showing the proportions of Cav3.2 mRNA-labeled cells based on a cross-sectional area (left). The proportion of cells labeled by an anti-NF-H antibody is shown as a reference on the right side. Only labeled cells that included nuclei were subjected to area measurements.</p