1. Copyright © 2010 American Medical Association. All rights reserved.
From: Autoantibody-Mediated Dysfunction of Sympathetic Neurons in Guillain-Barré Syndrome
Arch Neurol. 2010;67(2): doi: /archneurol
Figure Legend:
Staining properties, cell viability, and noradrenaline synthesis of sympathetic neurons during treatment with IgG from patients with Guillain-Barré syndrome (GBS) and control patients. A, Staining of sympathetic neurons with GBS-IgG (green) and antibody against tyrosine hydroxylase (red, bar = 50 μm). Cell nuclei are labeled with 4′,6-diamidino-2-phenylindol (DAPI). The IgG from controls and patients with GBS stained predominantly the cell bodies of rodent sympathetic neurons (see the Table for summary of staining pattern). B, Survival of sympathetic neurons in the presence of different human IgG. Treatment with GBS-IgG or control IgG does not affect average cell viability after 24 hours, as determined by fluorescence-based cell viability assay. C, Average change of noradrenaline levels in supernatants of sympathetic neurons 24 hours after treatment with GBS-IgG or control IgG. Noradrenaline levels declined in cultures treated with control IgG, but not in those exposed to GBS-IgG. CIDP indicates chronic inflammatory demyelinating polyradiculoneuropathy; RRMS, relapsing-remitting multiple sclerosis; *P < .05; error bars, standard error of the mean.
Date of download: 11/7/2017
Copyright © 2010 American Medical Association. All rights reserved.

2. Copyright © 2010 American Medical Association. All rights reserved.
From: Autoantibody-Mediated Dysfunction of Sympathetic Neurons in Guillain-Barré Syndrome
Arch Neurol. 2010;67(2): doi: /archneurol
Figure Legend:
Tyrosine hydroxylase expression of sympathetic neurons. A, Mean expression of tyrosine hydroxylase messenger RNA (mRNA) in sympathetic neurons in the presence of different IgG and after treatment with intravenous immunoglobulins (IVIg). Guillain-Barré syndrome (GBS)-IgG increased the expression of tyrosine hydroxylase mRNA in sympathetic neurons. Prior treatment of cells with IVIg prevented the increase of tyrosine hydroxylase mRNA (IVIg and GBS-IgG), in contrast to preincubation of IVIg and GBS-IgG before cell exposure. B, Western blot of cell lysates revealed increased tyrosine hydroxylase expression in sympathetic neurons exposed to GBS-IgG. C, Densitometric analysis of Western blot tyrosine hydroxylase bands in relation to protein concentration as determined by actin band intensity confirms mRNA data. Error bars indicate standard error of the mean; *P < .05; AU, arbitrary units; CIDP, chronic inflammatory demyelinating polyradiculoneuropathy; PBS, phosphate-buffered saline; preinc, preincubated; RRMS, relapsing-remitting multiple sclerosis.
Date of download: 11/7/2017
Copyright © 2010 American Medical Association. All rights reserved.

3. Copyright © 2010 American Medical Association. All rights reserved.
From: Autoantibody-Mediated Dysfunction of Sympathetic Neurons in Guillain-Barré Syndrome
Arch Neurol. 2010;67(2): doi: /archneurol
Figure Legend:
Guillain-Barré syndrome (GBS)-IgG and the beat rate of cardiomyocytes innervated by sympathetic neurons. A, Cocultures of sympathetic neurons and cardiomyocytes stained against tyrosine hydroxylase (red) and α-actinin (green), a marker to visualize cardiomyocytes (bar = 250 μm). B, Staining against tyrosine hydroxylase (green) and synaptophysin (red), a presynaptic marker for synaptic vesicle glycoprotein confirms synapse formation in cocultures of sympathetic neurons and cardiomyocytes (indirectly labeled with 4′,6-diamidino-2-phenylindol [bar = 50 μm]). C, Fluorescence-staining (left) and light microscopy (right) of a representative cluster of cardiomyocytes and innervating sympathetic neurons. Cells are stained with antibodies against tyrosine hydroxylase (red) and α-actinin (green) (bar = 100 μm). Arrowheads indicate the cell bodies of innervating sympathetic neurons. (A video of the cell cluster on the right is available here). D, Analysis of the average cardiomyocyte beat rate of cardiomyocytes without sympathetic neurons. The mean beat rate is not altered in cardiomyocytes, which were exposed to GBS-IgG or relapsing-remitting multiple sclerosis (RRMS)-IgG. E, Cardiomyocyte beat rate in cocultures of sympathetic neurons and cardiomyocytes, incubated with GBS-IgG and RRMS-IgG. F, Mean cardiomyocyte beat rate in cocultures of sympathetic neurons and cardiomyocytes, incubated with GBS-IgG, with GBS-IgG after preincubation with intravenous immunoglobulin (IVIg), or with GBS-IgG after preincubation with prazosin and propranolol. Error bars indicate standard error of the mean. *P < .05.
Date of download: 11/7/2017
Copyright © 2010 American Medical Association. All rights reserved.