5-hydroxymethylcytosine (5hmC) is considered as a novel
DNA modification and plays an important role in cancer, stem cells, and developmental diseases. In this study, we demonstrated the existence of
RNA 5hmC modification in mouse brain
RNA by using a dot blot analysis method. Our data indicated that 5hmC modification in
RNA samples was less than that in
DNA samples. Further, we optimized the conditions for 5hmC detection in
RNA samples such as
DNase treatment, denature
reagents, denature time, sample air-dry time, and the cross-linking time between
RNA and membrane. Our results demonstrated that
DNase treatment and denature
reagents were two important factors that affected the 5hmC detection in
RNA samples. By using the optimal conditions for
RNA 5hmC detection, we found that the brainstem, the hippocampus, and the cerebellum had high levels of 5hmC modification and 5mC modification in
RNA. Finally, we found that
RNA 5hmC modification decreased in
MPTP-induced
Parkinson's disease model in mice. These suggest that 5hmC modification in
RNA might play an important regulative role on
protein or
microRNA expression in these brain tissues. Because
DNA 5hmC modification plays an important role in neural differentiation and development as well as neurological diseases, the significance of 5hmC modification in
RNA in different neurological diseases needs further investigation. In summary, our study demonstrated for the first time the abundance of 5hmC modification in brain
RNA by using a dot blot analysis method and proved that dot blot analysis is a useful method for 5hmC detection in
RNA samples.