Table 2.

Shiga toxin and/or LPS administration model with CNS descriptions

Ref.	Animal (a)	toxin	CNS (b)	Gb3/Stx binding	Histopathology (c) (d) (e)	imaging	IHC (f)	Injection route (g)	Other assays
[66]	baboon (h)	Stx1	n.d.	n.d.	EM: mye deg, peri ede, large neu and glia deg, cap normal	n.d.	n.d.	i.v.
[67]	baboon	Stx1	yes	n.d.	n.d.	n.d.	n.d.	i.v.
[39]	weaned YL pigs	Stx2e	yes	n.d.	Gross: Sc ede, CL ede and hrrg HE: CL hrrg but not ede or cap nec, no lesions in CR, BS, thalamus,	n.d.	n.d.	i.v.
[40]	weaned YL pigs	Sup (i) Stx2e	yes	n.d.	HE: CL, sub ede, hrrg, cap nec HE: MB, fib nec, peri eos	n.d.	n.d.	i.v.
[41]	NZW rabbit	Stx1	yes	TLC-Stx1 over lay of CL, BS and Sc (+) at LacCer (j) position	HE: BS, Sc, CL cap narrowed, peri ede, cap damage and fib, Purkinje decreased	n.d.	n.d.	i.v.
[42]	NZW rabbit	Stx1	yes	125I-Stx1 tissue distribution high in cecum, brain, small intestine, colon, Sc	HE: cerv Sc hrrg, inf, ede, fib in cap, lum Sc fib cap, pyk cap	n.d.	Anti-Stx (+)in Sc cap	i.v.
[52]	JW rabbit	Stx2	yes	n.d.	EM: mye deg, axo normal	MRI: V3 (24 h), later BS, cc, lateral amy, CL vermis	Immuno EM-DAB (k): anti-Stx2 (+) at luminal side of cap, deg mye	i.v.	Tracer
[47]	JW rabbit	Stx2	Yes	n.d.	CR ctx, CL ctx, Sc, pyk neu, str neu not affected, inf in MB and CL	n.d.	Anti-Stx2 IHC (+)cap and sub	i.v. and i.t.	Stx2 ↑ in CSF
[65]	JW rabbit	Stx2	n.d.	n.d.	n.d.	MRI: CL at 82 h	n.d.	i.v.	Stx2 ↑ in CSF
[65]	JW rabbit	Stx2	n.d.	n.d.	n.d.	MRI: the rear of CL (contact w CSF) at 48 h	n.d.	i.t.	n.d.
[68]	JW rabbit	Stx2	yes	n.d.	n.d.	MRI: BS and cerv Sc dorsal close to death	n.d.	i.v.	Barorefl ex function
[69]	JW rabbit	Stx1	n.d.	n.d.	n.d.	MRI: MB, BS and Sc ede, cerv Sc dorsal hematoma	n.d.	i.v.	Stx1 ↑ slightly in CSF
[54]	JW rabbit young	Stx2	yes	n.d.	HE: inf CL, myo thickening, fib, pyk and fragmented, pons myo nec	n.d.	Anti-Stx2 IHC(+) in pons cap and myo, Anti-ssDNA IHC small number cap (+),IHC caspase-3 &-9 small number (+)	i.v.
[43]	JW rabbit	Stx2	yes	Anti-Gb3 (+) in cap lum Sc	HE: inf lum Sc with hrrg and fib cap, str neu pyk, hippos pyr neu apop	n.d.	TUNEL + in hippo (DG, CA1, CR ctx neu, pons glia, cap IHC: Ib4↑ (microglia activated) in lum Sc and thalamus	i.v.	qRT-PCR: TNFa↑ and IFNb ↑
[70]	JW rabbit	Stx2	n.d.	n.d.	n.d.	MRI: Gd leak (↑permeability)	n.d.	i.v.
[48]	DB rabbit	Stx2	yes	n.d.	HE: neu deg in the BS	n.d.	n.d.	i.v.
[49]	SD rat	Stx2	n.d.	n.d.	n.d.	n.d.	Anti-Stx IHC (+)peri	i.p.
[49]	SD rat	Stx2	n.d.	n.d.	EM: irregular shape neu, mye deg, hypertrophic axo, astro phago mye, neu apop, deg, vacuol, peri astro ede, non-peri astro gliosis, oligo pathologic	n.d.	Anti-Stx2 (+) in neu, immunoEM-DAB: anti-Stx2 (+) in neu fibers and astro nucleus	i.c.v.
[58]	SD rat	Stx2	n.d.	n.d.	n.d.	n.d.	Stx2 (+) in anterior hippo astro, ips hippo Stx2(+)astro and neu, cont hippo Stx2(+)neuropils	i.c.v.	Str and CR ctx neu↓, cap↓ NADPH-d/NOS activity
[57]	SD rat	Stx2	yes	Anti-Gb3↑ CA1, str neu	n.d.	n.d.	Anti-Stx2↑ MAP2 ↑ CA1 neu. Anti-bax ↑neu (inner ctx, CA1, subV dorsal str, hypothala mic peri	i.c.v.
[50]	SD rat	Sup Stx2	n.d.	n.d.	Nissle: neu pyk hypertrohy axo, ede ctx, subV CL.	n.d.	Anti-AQP4↓ cp	i.p.
[35]	ICR mouse	Stx2	yes	n.d.	n.d.	n.d.	Immune-EM-DAB: anti-Stx2 (+) mye deg, lyso pyr ctx	i.p.	Tracer
[44]	C57BL/6 mouse	Stx2	yes	n.d.	HE: pyk oligo astro nuclei, hrrg sub	n.d.	n.d.	i.v.
[44]	C57BL/6 mouse	Stx2 + LPS	yes	n.d.	HE: hrrg BS sub severe than Stx2 alone	n.d.	n.d.	i.v.
[45]	ICR mouse	Stx2	n.d.	n.d.	HE: congestion CL and hippo, hrrg CL, neu and glia normal	n.d.	anti-Stx2 IHC (+) in RBC and cap CL, MB and thalamus, anti-Stx2 IHC (−) in neu, glia	i.v.
[63]	C57BL/6 a4galt−/−	Stx1 and Stx2	Not suscep tible to Stx1	Anti-Gb3 became negative in cap	n.d.	n.d.	n.d.	Not specified
[53]	C57BL/6 mouse	Stx2	yes	Anti-Gb3 (+) in neu mouse and human Sc, human cap	EM: glia lamellipodia-like foot process interrupts synapse at motor neu of lum Sc		Immuno-gold EM (l): anti-Gb3 and anti-Stx2 double positive in motor neu of lum Sc	i.p.
[51]	NIH mouse	Stx2	n.d.	n.d.	EM: neu, astro and peri ede, synaptic disruption, oligo defect			i.v.	Behavioral motor test +
[61]	Human	n.a.	n.a.	DRG (m), Stx1 binding (+) neu and cap	n.a.	n.a.	n.a.	n.a.
[61]	Rabbit	n.a.	n.a.	DRG, Stx1 binding (+) neu and cap	n.a.	n.a.	n.a.	n.a.
[61]	rat	n.a.	n.a.	DRG, Stx1 binding (+) neu	n.a.	n.a.	n.a.	n.a.
[62]	Human	n.a.	n.a.	DRG, anti-Gb3 and Stx1 binding (+) neu and cap	n.a.	n.a.	n.a.	n.a.
[62]	rabbit	n.a.	n.a.	DRG, anti-Gb3 and Stx1 binding (+) neu and cap	n.a.	n.a.	n.a.	n.a.
[62]	Rat	n.a.	n.a.	DRG, anti-Gb3 and Stx1 binding (+) neu	n.a.	n.a.	n.a.	n.a.
[62]	mouse	n.a.	n.a.	DRG, anti-Gb3 and Stx1 binding (+) neu	n.a.	n.a.	n.a.	n.a.
[60]	C57BL/6 mouse	n.a.	n.a.	Anti-Gb3 (+) neu at olf, CR ctx, str, hippo, hypothalamus, CVOs, CL, MO, Sc V3 ependyma	n.a.	n.a.	n.a.	n.a.

^(a)Animal keys: YL (Yorkshire-Landrace), NZW (New Zealand White), JW (Japanese White), DB (Dutch Belted), SD (Spraque-Dawley)

^(b)Detailed CNS symptoms are summarized in Table 3.

^(c)Histopathology analysis keys are Gross (gross observation in non-stained tissue), HE (hematoxylin-esosin stain that stains cytoplasm in pink and nucleus blue, light microscopic findings (LM)), PAS (Periodic acid-Schiff stain that detects polysaccharides, glycoproteins and glycolipid, LM), LFB (Luxol fast blue stain that stains myelin in blue, LM), EM (electron microscopic findings)

^(d)CNS regions and cell type abbreviations are CR (cerebrum), ctx (cortex), hippo (hippocampus), DG (dentate gyrus), str (striatum and other basal ganglia), CL (cerebellum), MO (medulla oblongata), MB (midbrain), BS (brain stem is used where midbrain, pons or medulla oblongata are not specified), Sc (spinal cord), cerv (cervical), tho (thoracic), lum (lumbaris), sub (subarachinoid space),, Histopathologic feature abbreviations are cap (endothelial cells or capillaries), inf (infarction), hrrg (hemorrhage), fib (fibrin deposition), nec (necrosis), swl (swelling), peri (perivascular), myo (myocytes), apop (apoptotic), mono (monocytes), mye (myelin), deg (degeneration), pyk (pyknotic nuclei), prolif (proliferation/hyperplasia), ede (edema), pyr (pyramidal neuron), inflt (infiltration of blood cells to parenchyma), neu (neuron), Purkinje (Purkinje cells are large neurons in CL), V3 (third ventricle), cc (corpus callosum), amy (amygdala), ips (ipsilateral, injection side of brain), cont (contlateral, opposite of injection side), subV (subventricular region), cp (choroid plexus), CVO (circumventricular organs)

^(e)Histopathologic feature abbreviations are cap (endothelial cells or capillaries), inf (infarction), hrrg (hemorrhage), fib (fibrin deposition), nec (necrosis), swl (swelling), peri (perivascular), myo (myocytes), apop (apoptotic), mono (monocytes), mye (myelin), deg (degeneration), pyk (pyknotic nuclei), prolif (proliferation/hyperplasia), ede (edema), pyr (pyramidal neuron), inflt (infiltration of blood cells to parenchyma), neu (neuron), Purkinje (Purkinje cells are large neurons in CL), glia (glial cells such as astrocytes, microglia and oligodendrocytes), eos (eosinophilic globules, deposits), axo (axon, axoplasm), astro (astrocyte), oligo (oligodendrocyte), phago (phagocytosis), lyso (lysosome), RBC (red blood cells)

^(f)IHC = immunohistochemistry, immunodetection of the target in the tissue sections

^(g)Injection route abbreviations: i.v. (intravenous), i.t. (intrathecal, injection from cysterna magna that makes it possible to inject into cerebrospinal fluid (CSF), i.p. (intraperitoneal), i.c.v. (intracerebroventricular injection that inject solution directly into CNS parenchyma of cerebral cortex/ventricle).

^(h)Baboon in this chart is Papio c. cynocephalus, or Papio c. Anubis

⁽ⁱ⁾Sup = E. coli culture supernatant

^(j)LacCer = lactsylceramide, adding galastose to LacCer completes Gb₃.

^(k)Immuno-EM-DAB: immunodetection of the target with 3,3′-diaminobenzidine (DAB) deposition by EM

^(l)Immunogold EM: immunodetection of the target with 5–10 nm gold particle allows precise localization as well as double labeling.

^(m)DRG = dorsal root ganglion, a peripheral nervous system structure consists of sensory neurons and other cell types.