Figure 1

Characterization of engineered changes in the ADK expression profile during the postnatal development of Adk-tg cerebellum. (A) Expression profile of ADK-L and ADK-S proteins in the cerebellum from Adk-tg and WT mice at different postnatal developmental stages. Representative cropped blots show ADK (L and S) isoform expression at the cerebellar developmental stages P5, P7, P9, P15, P21, with the long ADK-L isoform appearing as an upper band and the short ADK-S isoform as the lower band. The WT cerebellum showed increased expression of both ADK isoforms (S and L) during developmental maturation. In contrast, the Adk-tg cerebellum was characterized by constant expression of the Adk-S transgene at all developmental stages. (B) Quantitative analysis of the expression profile of ADK-S in the developing cerebellum of Adk-tg and WT mice. In WT cerebellum, ADK-S expression increased progressively and significantly from P5 to P21, while constant levels (~ 1.5-fold increase of ADK-S compared to WT at P5) were found during the development of the Adk-tg cerebellum. (C) Quantitative analysis of the expression profile of ADK-L in the developing cerebellum of Adk-tg and WT mice. In WT cerebellum, ADK-L expression increased progressively and significantly from P5 to P21, whereas ADK-L expression was absent in the cerebellum of Adk-tg mice. Quantitative analysis was performed using Image J V. 1.52 software and expressed as the ratio of optical densities of ADK expression levels normalized to the internal standard beta-tubulin. (D) Immunostaining of ADK (red) and Hoechst (blue) in WT and Adk-tg mice. White arrows show colocalization of Hoechst-positive cells nuclei with Adk-L in the molecular layer (ML) and internal granular layer (IGL). Scale bar is 50 and 25 µm. All values are presented as mean ± SEM (n = 3–4). Two-way ANOVA with Tukey's multiple comparison post-hoc test (*p < 0.05, **p < 0.01, and ***p < 0.001 for significance).