Figure 4

Immunolocalization of NCX isoforms in primary cultured TG neurons. (A, B, and C) NCX immunoreactivity (red) and nuclei (blue). Intense immunoreactivity was observed for NCX1 (A), NCX2 (B), and NCX3 (C) in TG neurons. (D) Positive control for Pan neuronal marker in TG neurons. Pan neuronal marker includes anti-NeuN, anti-MAP2, anti-βIIItubulin, and anti-neurofilament 200 (NF-H) antibodies. (E, F, and G) Double immunofluorescence staining of NCX isoforms (green) and Pan neuronal marker (red). All NCX isoforms were localized on the somata, dendrites, axons, and perinuclear region of TG neurons. (H) Positive immunoreactivity to NF-H as an A-neuron marker in TG neurons. (I, J, and K) Double staining of NCX isoforms (red) and NF-H (green). (L) Positive immunoreactivity to isolectin B4 (IB4; as a non-peptidergic C-neuron marker) in TG neurons. (M, N, and O) Double staining of NCX isoforms (red) and IB4 (green). (P) Positive immunoreactivity to calcitonin gene-related peptide (CGRP; as a peptidergic C-neuron marker) in TG neurons. (Q, R, and S) Double staining of NCX isoforms (red) and CGRP (green). Intense immunoreactivity for all of NCX isoforms was observed on A-, non-peptidergic C-, and peptidergic C-neurons. No fluorescence was detected in the negative control (data not shown). Scale bar: 20 μm.