Retrograde labelling of sub-coeruleus neurones using an adenoviral vector.
An adenoviral vector containing PRSx8-Kir2.1-IRES-EGFP was injected into the lumbar dorsal horn of the rat. Retrograde labelling is observed in pontospinal regions (A5-A7). Although EGFP expression is sparse, it is possible to detect expression by using anti-EGFP (RED, 1:4000, A11122, Invitrogen, 1:1000, Biotin SP anti-rabbit, 711-066-152, Jackson ImmunoResearch and strept-avidin Cy3, 1:1000, Sigma). These retrogradely labelled neurones were DβH positive (BLUE , 1:5000, MAB308, chemicon, 1:200, AMCA conjugated anti-mouse, 715-156-150, Jackson ImmunoResearch). Key: 4 – overlay; scale bars 50μM.