Fig 1: Co-labeled cells express choline acetyltransferase and vesicular glutamate transporter 2. Most neurons labeled with dextran dyes coming from the carotid chemoreceptors (CB, Alexa 488, green) and pulmonary nociceptors (cell bodies in the nodose ganglion, ND, Alexa 647, red) in the nucleus tractus solitarius (NTS) and nucleus ambiguus (NA) were labeled with vesicular glutamate transporter 2 (Vglut2, Cy3, yellow) and in close relation to choline acetyltransferase (ChAT, Dylight 405, magenta) stained cells indicative of an excitatory phenotype. Bottom right inset shows a lack of staining but with tracers remaining, when the primary antibodies were omitted to serve as no primary control staining. ×40, scale bar = 20 μm. Images taken with Leica SP8 Confocal Microscope.
Fig 2: Expression profiles of α-syn in lesions of patients with FCD IIb and TSC. (A) Moderate-to-strong α-syn immunoreactivity was observed in the neuropils of CTX but was neglectable in the neuronal cytoplasm. (B) Representative image showing weak α-syn signal in the neuropils of FCD IIb lesions and moderate α-syn immunoreactivity in DN (double arrow) and BC (arrow) of FCD IIb lesions. (C) Strong but less of α-syn positive signal was observed in the neuropils of TSC lesions, and α-syn immunoreactivity was barely detected in DN (arrow) and GC (double arrow) of TSC lesions. (D) Statistical analysis results showing that the average OD value of α-syn was reduced in neuropils of both FCD IIb and TSC lesions (***p < 0.001, unpaired two-tailed t-test, n = 7, 9, 8 specimens for each group). (E) Representative images showing α-syn was expressed in vimentin-positive BC in FCD IIb lesions (arrows). (F) α-syn was barely expressed in VGLUT2-positive excitatory neurons in CTX. (G) Representative image showing the co-expression of α-syn and VGLUT2 was intensified in the neuropils, DN (double arrow), and BC (arrow) in FCD IIb lesions. (H) The co-expression of α-syn and VGLUT2 was intensified in the neuropils of TSC lesions as well but not observed in DN and BC. (I) α-syn was not detected in vimentin-positive GCs in TSC lesions. (J) Representative image showing α-syn was barely expressed in VGAT-positive neurons in CTX. (K) Representative image showing significant co-expression of α-syn and VGAT in DN (double arrow) and BC (arrow) of FCD IIb lesions. (L) The co-expression of α-syn and VGAT was intensified in the neuropils of TSC lesions but not observed in DN and GC of TSC lesions. (M) Representative image showing slightly α-syn expression in a NeuN-positive neuron and nearby neuropils in FCD IIb lesions (arrow). (N–P) Representative images showing α-syn was not detected in GFAP-positive astrocytes ((N), arrows), Iba1-positive microglia (O, arrow), and Olig2-positive oligodendrocytes ((P), arrows) in FCD IIb lesions. (Q–T) Representative images showing α-syn was not expressed in NeuN-positive neurons in TSC lesions ((Q), arrow), GFAP-positive ((R), arrows), Iba1-positive microglia ((S), arrows), and Olig2-positive oligodendrocytes in TSC lesions ((T), arrow). Scale bars: 50 μm for the upper panel of (A–C), 20 μm for the bottom panel of (A–C), and 10 μm for images (E–T).
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