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. 2013 Mar 20;33(12):5227–5240. doi: 10.1523/JNEUROSCI.4452-12.2013

Figure 1.

Figure 1.

Nerve ligation induces behavioral allodynia and spinal kalirin expression. A, Spinal nerve ligation [SNL IPSI; **p < 0.01 vs sham operation (sham IPSI), n = 7], but not the sham operation, statistically decreased the mechanical threshold of the ipsilateral hindpaw when tested on days 1, 3, 7, 14, and 21 compared with the presurgery controls (day −1; ##p < 0.01 vs day −1, n = 7). In contrast, neither the sham operation (sham CONTRA) nor the spinal nerve ligation (SNL CONTRA) affected the mechanical threshold of the contralateral hindpaw tested at these time points (p > 0.05 vs day −1). B, Representative Western blots of kalirin expression after spinal nerve ligation. The dorsal horn lysates were blotted for kalirin, and the level of GAPDH was used as a loading control. IB, immunoblotting. C, Statistical analysis revealed that spinal nerve ligation statistically upregulated the level of kalirin in the ipsilateral dorsal horn when compared with the presurgery control [*p < 0.05, **p < 0.01 vs day −1, n = 7; ##p < 0.01 vs sham operation (sham IPSI), n = 7]. In contrast, the sham operation exerted no effect on kalirin expression quantified at identical time points (p > 0.05 vs day −1, n = 7). The data show the mean ± SEM. D, Spinal nerve ligation (II, SNL7D), but not the sham operation (I, sham 7D), notably enhanced the kalirin immunoreactivity (red) in the ipsilateral dorsal horn (IPSI) of spinal sections obtained on day 7 after surgery when compared with the contralateral dorsal horn (CONTRA). In contrast, the immunoreactivity of NeuN (III, green) was not different between the ipsilateral and the contralateral dorsal horns. Double-staining images revealed that the nerve ligation-enhanced kalirin immunoreactivity was colabeled with NeuN (IV, yellow). Amplified images of IV showed that NeuN (V, green), but not OX-42 (VI, green) or GFAP (VII, green), was colocalized with the enhanced kalirin immunoreactivity (V, yellow). Scale bar, 50 μm; thickness, 50 μm. Each of these immunofluorescence images was replicated in seven sample preparations with similar results each time.