Neurotransmitter metabolites (dihydroxyphenylacetic acid [DOPAC], homovanillic acid [HVA], and 5-hydroxyindoleacetic acid [5-HIAA]) in CSF of 10 healthy dogs were evaluated with reverse-phase high-pressure liquid chromatography and electrochemical detection. Neurotransmitter metabolite concentrations determined in CSF collected from the cisterna magna were compared with those values in CSF collected from the lumbar dorsal subarachnoid space. Amounts of DOPAC (P = 0.0444), HVA (P = 0.0001), and 5-HIAA (P = 0.0316) were significantly lower in lumbar spinal fluid compared with those values in the cervical spinal fluid. Metabolite concentrations in cervical and lumbar CSF were: DOPAC = 2.81 +/- 0.73 ng/ml of CSF and 1.28 +/- 0.57 ng/ml; HVA = 98.29 +/- 12.42 ng/ml and 4.68 +/- 1.61 ng/ml; and 5-HIAA = 46.29 +/- 8.17 ng/ml and 36.96 +/- 4.07 ng/ml, respectively. Cytologic evaluations of cervical and lumbar CSF revealed a similar concentration of 3 +/- 1 WBC/microliters in both fluids. A significant (P = 0.0002) difference in protein concentration between the 2 regions was observed, with 16.1 +/- 1.8 mg of protein/dl in the cervical CSF and 27.2 +/- 2.3 mg of protein/dl in the lumbar CSF. Between the cisterna magna and lumbar dorsal subarachnoid space of dogs, a rostrocaudal gradient existed for neurotransmitter metabolites, and a caudorostral gradient existed for protein.