Progressive degeneration of nigrostriatal dopamine neurons following intrastriatal terminal lesions with 6-hydroxydopamine: a combined retrograde tracing and immunocytochemical study in the rat

Abstract

In order to develop a rodent model displaying a progressive degeneration of the dopamine neurons of the substantia nigra, we bilaterally injected the tracer substance FluoroGold into the terminal field of the nigrostriatal projection, i.e. the striatum. One week later, rats received unilateral injections of 20 micrograms 6-hydroxydopamine into one of the two striatal tracer deposits. Groups of animals were killed one, two, four, eight and 16 weeks later. Ipsilateral to the lesion there was a progressive loss of FluoroGold-labelled nigral cells, with cell counts dropping from 96% of the contralateral side at one week to 59% at two weeks, 35% at four weeks, 23% at eight weeks and down to 15% at 16 weeks. Labelled nigral neurons ipsilateral to the lesion showed a moderate to marked atrophy at all investigated time points. The number of tyrosine hydroxylase-immunoreactive cells was decreased to 83% of contralateral at one week, 39% at two weeks, 44% at four weeks, 34% at eight weeks and 52% at 16 weeks postlesion. Rhodamine fluorescence immunocytochemistry showed that the proportion of surviving ipsilateral fluorogold-labelled cells displaying immunoreactivity for tyrosine hydroxylase was 69% at one week postlesion, 51% at two weeks, 63% at four weeks, 69% at eight weeks and 76% at 16 weeks. We conclude that injection of 6-hydroxydopamine into the terminal field of nigral dopaminergic neurons causes a progressive degeneration of these cells, starting between one and two weeks after lesion and continuing over eight to 16 weeks. This degeneration is preceded, and accompanied by, cellular atrophy and a partial loss of marker enzyme expression, thus yielding an animal model which mimics the degenerative processes in Parkinson's disease more closely than the animal models available so far. The present model may be helpful in investigating the in vivo effects of putative neuroprotective agents and neurotrophic factors.