IL-1β and IL-1ra mRNA in CD68 positive cells.

<p>Expression of IL-1β and IL-1ra mRNA in CD68 positive activated macrophages/microglial cells during the first disease phase of EAE. <b>(A, B)</b> IL-1β mRNA (black grains) combined with CD68 immunoreactivity (brown) in the vestibular nucleus of the brain stem. <b>(C, D</b>) IL-1ra mRNA (black grains) combined with CD68 immunoreactivity in the spinal cord. Scale bars = 20 µm.</p

IL-1β and IL-1ra mRNA in first phase and remission of cr-EAE.

<p>Film autoradiographs of affected brain regions in cr-EAE rats after <i>in situ</i> hybridization for IL-1β (left pannels) or IL-1ra (middle pannels) or with sense probes (right panels). Upper and middle pannels are taken from animals in first disease phase, lower panels from animals in remission. <b>(A, B)</b> clusters of IL-1β and IL-1ra mRNA in triangular septal nucleus (TS) and ventral hippocampal commissure (vhc); <b>(D, E)</b> clusters of IL-1β and IL-1ra mRNA in stria medullaris of the thalamus (sm) and habenular nuclei (Hb), and <b>(G, H)</b> IL-1β and IL-1ra mRNA in optic tract (OT). <b>(C, F)</b> sections hybridized with a sense probe for IL-1β, (<b>I</b>) section hybridized with a sense probe for IL-1ra. Scale bar (<b>A–I)</b> = 2 mm.</p

Distribution of IL-1β and IL-1ra mRNA in affected brain regions of cr-EAE rats.

<p>Grey areas represent localization of IL-1β and IL-1ra mRNA in the <b>(1)</b> triangular septal nucleus, <b>(2)</b> septofimbrial nucleus, <b>(3)</b> ventral hippocampal commissure, <b>(4)</b> subfornical organ, <b>(5)</b> paraventricular thalamic nucleus, <b>(6)</b> stria medullaris of the thalamus, <b>(7)</b> 3^rd ventricle, <b>(8)</b> medial preoptic area, <b>(9)</b> nucleus of diagonal band, <b>(10)</b> suprachiasmatic nucleus, <b>(11)</b> supra optic hypothalamic nucleus, <b>(12)</b> optic chiasm, <b>(13)</b> medial habenular nucleus, <b>(14)</b> lateral habenular nucleus, <b>(15)</b> intermediodorsal thalamic nucleus, <b>(16)</b> lateral thalamic nuclei, <b>(17)</b> optic tract, <b>(18)</b> habenular commissure, <b>(19)</b> cerebellar lobules, <b>(20)</b> cerebellar peduncles, <b>(21)</b> parabrachial nucleus, <b>(22)</b> central grey pons, <b>(23)</b> trigeminal nucleus, <b>(24)</b> cochlear nucleus, <b>(25)</b> predorsal bundle, <b>(26)</b> pontine reticular nucleus, <b>(27)</b> vestibular nucleus, and <b>(28)</b> spinal trigeminal tract.</p

Histopathological features of CNS lesions during the first disease phase of cr-EAE.

<p>Left panels: PLP immunoreactivity. Middle panels: CD68 immunoreactivity. Right panels: CD3 immunoreactivity during the first disease phase of cr-EAE in (<b>A–C</b>) the trigeminal tract (WM), and (<b>D–F</b>) the trigeminal nucleus (GM). Scale bar = 60 µm.</p

Semi-quantitative RT-PCR analysis of IL-1β and IL-1ra mRNA.

<p>Q-PCR of IL-1β and IL-1ra mRNA in brainstem and spinal cord during the first phase and relapse of cr-EAE. Data represent mean ± S.E.M. (n = 4) and are expressed relative to GAPDH mRNA. *<i>P</i><0.05 vs first phase.</p

IL-1β expression, CD68 immunoreactivity and Oil-Red O staining.

<p>Expression of mRNA for IL-1β, presence of activated macrophages/microglial cells, and lipid fragmentation. All pictures were taken from animals at the first phase of cr-EAE. <b>(A)</b> IL-1β mRNA in cerebellar lobule. <b>(B)</b> Oil-Red O in cerebellar lobule. Arrows indicate lipid laden cells. <b>(C)</b> CD68 in cerebellar lobule. <b>(D)</b> IL-1β mRNA in the habenula. <b>(E)</b> Absence of IL-1β mRNA in the habenula of a control animal <b>(F)</b> Oil-Red O in the habenula. Sections are counterstained with Cresyl Violet <b>(A, D)</b> or with Mayer's hematoxylin <b>(B, E, F)</b> D3V: dorsal third ventricle, Hb habenula nuclei, LPMR: lateral posterior thalamic nucleus, sm: stria medullaris. Scale bars = 20 µm.</p

CD68 positive cells in the brain stem and the spinal cord during cr-EAE.

<p>Left panels: first phase of disease, middle panels: remission, and right panels: relapse. <b>(A–C)</b> CD68 positive cells in the brain stem, and <b>(D–F)</b> in the spinal cord. Note the difference in the morphology of CD68 positive cells in the brain stem and spinal cord during the relapse phase. Scale bar = 20 µm. Arrows in <b>C</b> indicate ramified CD68 positive cells; arrowheads in <b>F</b> indicated cells with an amoeboid morphology.</p

Neurological symptoms during cr-EAE.

<p>Male dark agouti rats were immunized with MOG and neurological symptoms were monitored daily. Data represent mean and S.E.M. of neurological scores. Note the typical multiphasic course of the disease with a first phase followed by complete or partial remission and subsequent relapse.</p

Oligonucleotide primers used for cDNA amplification.

<p>Oligonucleotide primers used for cDNA amplification.</p

TG activity in spinal cord post-mortem tissue derived from EAE animals treated with vehicle, BJJF078 or ERW1041E.

<p>(a-f) <i>In situ</i> TG activity was detected in spinal cord sections derived from (a,d) vehicle, (b,e) BJJF078 and (c,f) ERW1041E treated mice. (d-f) Co-incubation of the sections with the TG2 inhibitor Z006 diminished TG activity. Scale bar: 50 μm. n = 11 or 12 animals/group.</p