Differential vulnerabilities of substantia nigra catecholamine neurons to excitatory amino acid-induced degeneration in rat midbrain slices

Although differential vulnerability in different regions of the central nervous system is a characteristic feature of neurodegenerative disorders in vivo, its cellular basis is not well understood. In the present study we investigated whether catecholamine neurons in different regions of the substantia nigra (SN) are differentially vulnerable to excitatory amino acid-induced damage in a midbrain slice preparation. Rats were anesthetized by halothane inhalation and killed, the brain was rapidly removed, and 300-microm-thick midbrain slices were cut horizontally on a vibratome. The slices were incubated at 35 degrees C for 2 h in saline buffer containing either kainic acid (KA) or N-methyl-d-aspartate (NMDA) (10-50 microM). They were then fixed and cut into 30-microm sections that were coplanar with the horizontal slice. Individual catecholamine neurons were identified in these thin sections using an antibody to tyrosine hydroxylase coupled to diaminobenzidine. Catecholaminergic neurons in the dorsal and ventral tiers of the SN were readily identified by reference to an atlas of the distribution of catecholamine neurons in the horizontal plane. Using dendritic degeneration as a sensitive index of damage, and submaximal concentrations of KA and NMDA, we found that catecholamine neurons in the dorsal tier were more vulnerable than those in the ventral tier. For example, KA (10 microM) caused a significant reduction in the proportion of neurons with dendrites in the dorsal tier (from 60 to 34%) without altering the dendritic arbor of ventral tier neurons. After treatment with 50 microM KA, only 11% of dorsal tier neurons retained any dendrites while 45% of ventral tier neurons retained their dendrites. These differences were statistically significant (P<0.001). A similar differential vulnerability was apparent in slices treated with NMDA; neurons in the dorsal tier lost dendrites before detectable damage in the ventral tier. An understanding of the comparative anatomical, neurochemical, and physiological properties of vulnerable (dorsal tier) and resistant (ventral tier) catecholamine neurons in rat SN may provide significant insights into the mechanisms and treatment of neurodegenerative disorders involving catecholamine neurons.