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ANA_21966_sm_SupplFigure1.TIF34818KSupplementary Figure 1: Predominantly neuronal alpha-synuclein immunopositive inclusion bodies in the cerebral cortex, amygdala, hypothalamus, and thalamus (A) Allocortical entorhinal region: abundant alpha-synuclein immunopositive LBs in the deep layers V and VI (arrows). (B) Allocortical hippocampus: Severe affection of the CA1, CA2, and CA3 sectors and slight affection of the fascia dentata (FD) by alpha-synuclein immunopositive neuronal inclusions bodies. (C) Amygdala: Close-meshed network of alpha-synuclein immunopositive LBs and LNs in the central nucleus (CE) and markedly affected intercalated nucleus (IC). (D) Hypothalamus: Remarkable involvement of the tuberomammillary nucleus (TUM). (E) Dense mesh of alpha-synuclein immunopositive LBs and LNs in the intralaminar central medial (CEM) and cucullar (CU) thalamic nuclei, as well as in the anterodorsal nucleus (AD) of the thalamus. Note the additional slight involvement of the thalamic anteroprincipal (AP) and mediodorsal nuclei (MD). (F) Severe affection of the thalamic intralaminar central medial (CEM) nucleus and the paraventricular nuclei (PV). (G) Selective vulnerability of the islands of the intralaminar central lateral nucleus (CL) of the thalamus intermingled among the nearly unaffected nerve cells of the medial subnucleus of the pulvinar (PU m). (H) Dense network of LBs and LNs in the limitans-suprageniculate complex (LI-SG) lying at the border between the thalamus and the midbrain. Note the marked involvement of the pretectum (PR). (A-H: anti-alpha-synuclein immunocytochemistry, 100 μm PEG sections). Abbreviations: AD – Anterodorsal nucleus of the thalamus; AP – Anteroprincipal nucleus of the thalamus; CA1 – CA1 sector of the hippocampus; CA2 – CA2 sector of the hippocampus; CA3 – CA3 sector of the hippocampus; CE – Central nucleus of the amygdala; CEM – Central medial nucleus of the thalamus; CL – Central lateral nucleus of the thalamus; CU – Cucullar nucleus of the thalamus; FD – Fascia dentata; FO – Fornix; IC – Intercalate nucleus of the amygdala; LI-SG – Limitans-suprageniculate-complex of the thalamus; MD – Mediodorsal nucleus of the thalamus; PR – Pretectum; PU m – Pulvinar, medial subnucleus; PV – Paraventricular nuclei of the thalamus; TUM – Tuberomammillary nucleus of the hypothalamus; IV – Cortical layer IV; V – Cortical layer V; VI – Cortical
ANA_21966_sm_SupplFigure2.tif33247KSupplementary Figure 2: Alpha-synuclein immunopositive neuronal and oligodendroglial inclusions in subthalamic and brainstem nuclei, as well as alpha-synuclein immunopositive brainstem fiber tracts (A) Abundance of alpha-synuclein immunopositive neuronal (arrow) and oligodendroglial inclusions in the zona incerta (ZI) and slightly affected subthalamic nucleus (SU) (arrow). (B) High prevalence of alpha-synuclein immunopositive LBs (arrow), LNs, and coiled bodies in the substantia nigra (SN). Note the considerable affection of the red nucleus (RD). (C) Dense network of alpha-synuclein immunopositive LBs and LNs in the precerebellar reticulotegmental nucleus of the pons (RTTG) and markedly affected oral subnucleus of the pontine reticular formation (PNO). (D) Large amounts of LBs and LNs in the raphe magnus (RMG) and gigantocellular reticular nuclei (GI). Additional LBs (arrow) are present in the dorsal accessory subnucleus of the inferior olive (IOD). (E) Severe affection of the dorsal motor vagal (DMV) and solitary nuclei (SOL), as well as alpha-synuclein immunopositive vagal nerve (X). (F) Severe alpha-synuclein immunopositive inclusion body pathology in the intermediate reticular zone (IRZ) and the precerebellar lateral reticular nucleus (LRT). Note the alpha-synuclein immunopositive fibers of the dorsal spinocerebellar tract (arrows). Alpha-synuclein immunopositive (G) vestibular nerve (VIII) and (H) olivocerebellar fibers (arrowheads). (A-H: anti-alpha-synuclein immunocytochemistry, 100 μm PEG sections). Abbreviations: DMV – Dorsal motor vagal nucleus; ICP – Inferior cerebellar peduncle; IOD – Inferior olive, dorsal accessory subnucleus; IOP – Inferior olive, principal subnucleus; IRZ – Intermediate reticular zone; GI – Gigantocellular reticular nucleus; LRT – Lateral reticular nucleus; PNO – Pontine reticular formation, oral subnucleus; RD – Red nucleus; RMG – Raphe magnus nucleus; RTTG – Reticulotegmental nucleus of the pons; SN – Substantia Nigra; SOL – Solitary nuclei; SOL – Solitary tract; SU – Subthalamic nucleus; ZI – Zona incerta; VIII – Vestibular nerve; X – Vagal nerve
ANA_21966_sm_SupplFigure3.tif23712KSupplementary Figure 3: No colocalization of alpha-synuclein and tau pathologies in the dorsal raphe nucleus and intermediate reticular zone of the A30P patient Immunofluorescent images displaying concomitant (A) alpha-synuclein and (B) tau inclusion pathologies in the intermediate reticular zone of the A30P patient, which (C) do not colocalize. (D) The alpha-synuclein and (E) tau aggregates likewise (F) do not colocalize in the dorsal raphe nucleus of the A30P patient. (A-F: Immunofluorescence staining of alpha-synuclein inclusion pathology with alexa 568 chromogen (red) and tau inclusion pathology with alexa 488 (green), 100 μm PEG sections).
ANA_21966_sm_suppltext.doc59KSupplemental Material
ANA_21966_sm_Suppl_TABLE1.doc44KTable 1: Distribution and extent of alpha-synuclein immunopositive neuronal and oligodendroglial inclusions in the cerebral cortex, basal forebrain, basal ganglia and amygdala of the A30P index patient affected by both LBs and/or LNs and coiled bodies (L, Lewy bodies and/or Lewy neurites; C, coiled bodies; not discernible 0, mild +, marked ++, severe +++).
ANA_21966_sm_Suppl_TABLE2.doc44KTable 2: Distribution and extent of alpha-synuclein immunopositive neuronal and oligodendroglial inclusions in the thalamus and hypothalamus of the A30P index patient affected by both LBs and/or LNs (L, Lewy bodies and/or Lewy neurites; C, coiled bodies; not discernible 0, mild +, marked ++, severe +++).
ANA_21966_sm_Suppl_TABLE3.doc42KTable 3: Distribution and extent of alpha-synuclein immunopositive neuronal and oligodendroglial inclusions in subthalamic nuclei, the midbrain and pons of the A30P index patient affected by both LBs and/or LNs (L, Lewy bodies and/or Lewy neurites; C, coiled bodies; not discernible 0, mild +, marked ++, severe +++).
ANA_21966_sm_Suppl_TABLE4.doc36KTable 4: Distribution and extent of alpha-synuclein immunopositive neuronal and oligodendroglial inclusions in the medulla oblongata and cerebellum of the A30P index patient affected by both LBs and/or LNs (L, Lewy bodies and/or Lewy neurites; C, coiled bodies; not discernible 0, mild +, marked ++, severe +++).
ANA_21966_sm_Suppl_TABLE5.doc37KTable 5: Extent of alpha-synuclein immunoreactive fibers in the central nervous white matter components of the A30P index patient showing additional oligodendroglial inclusions (IF, immunoreactive fibers; C, coiled bodies; not discernible 0, mild +, marked ++, severe +++).
ANA_21966_sm_Suppl_TABLE6.doc29KTable 6: Clinical and pathological characteristics of brain donors used for biochemical studies.

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