Effect of Gamma Knife Radiosurgery on Rat Brain Sodium Channel Subunit mRNA Expression

Abstract

Several lines of evidence underscore a possible and delayed antiepileptic effect of Gamma Knife irradiation. This effect could be related to structural and molecular changes. Since voltage-gated Na⁺ channels (NaChs) play a crucial role in neuron excitability, we studied the effect of Gamma Knife (GK) irradiation on the distribution of Na⁺ channel (NaCh) subunit mRNAs in rat brains. A left side irradiation was performed in rats using a stereotactic device adapted for GK radiosurgery. A dose of 100 Gy was administered with the 4 mm collimator. The left dentate gyrus and thalamus coordinates were based on De Groot’s rat stereotactic atlas. The isodose curve distribution was calculated with the dose planning software used in Gamma Knife and superimposed on the target. Na⁺ channels α unit and mRNAs (subtype II and subtype III) expression was studied 1 hour, 30 days and 60 days later. We used non-radioactive in situ hybridization with subtype-specific digoxigenin-labeled cRNA probes. Labeling intensity was evaluated with a densitometric analysis of digitized images from the control side (right) and lesioned side (left) in each rat. No morphological changes were observed one hour after GK irradiation. 30 days later, the upper thalamic nuclei exhibited a few necrotic regions associated with gliosis. In contrast, no lesions were observed in the hippocampus. 60 days later, the necrotic region involving thalamic nuclei was enlarged. NaCh II and III mRNAs expression did not appear to be modified after GK at the three times studied here. In particular, neurons surrounding the GK necrosis continued to express high levels of NaCh mRNAs. Thus, regulation of NaCh II and III subtypes do not appear to explain the functional antiepileptic effect of GK.