Abstract:

Objective To investigate expression of Parkinson disease(PD) related protein PINK1 in rat brain. Methods To obtain the antiserum against human PINK1, two New Zealand white rabbits(2.5 kg) were immunized with recombinant Histag fusion human PINK1 protein; the antiserum was purified by salt fractionation, the specificity and sensitivity of the antiserum were verified by Western blotting. The location and distribution of the protein were further determined by immunohistochemistry. Results The human recombinant PINK1 gene was highly expressed as inclusions in prokaryotic expression system. After first salt fractionation, western blotting analysis demonstrated that the antiserum detected a 40 000 band in recombinant human PINK1 protein, a 40 000 band and a 66 000 band in rat brain homogenates which is anticipated. Immunohistochemical staining on normal adult rat brain sections showed that the PINK1immunoreactive substance presents in rat brain bulbus olfactorius, cortex, hippocampus, striatum, cerebellum, substantia nigra and red nucleus. Conclusion A polyclonal rabbit antiserum against human PINK1 was prepared and the immunoreactive PINK1 substance was widely distributed in rat CNS. The antiserum provided a useful experimental tool for profound studies on the tissue expression profile, intercellular location and biological function of PINK1.

Key words: PINK1, polyclonal antiserum, rat, recombinant protein