Carbon nanohorns (CNHs), closed cone-shaped cages of sp2-hybridized carbons, are a promising nanomaterial to improve carbon-fiber microelectrode (CFME) dues to their high specific surface area and edge planes, but few studies have tested their electrochemical properties. Here, we tested the dopamine detection at electrodeposited CNHs on CFME (CNH/CFME). The optimized concentration of CNHs in the deposition solution is 0.5 mg/mL, and the optimized electrodeposition waveform is 10 cycles of triangular waveform scanned from –1.0 V and +1.0 V at 50 mV/s. Using fast-scan cyclic voltammetry, the optimized CNH/CFME enhances dopamine peak current to 2.3 ± 0.2 times that of the CFME. To further increase the current, CNH/CFMEs were oxidized in NaOH (ox-CNH/CFME), which creates more defects and surface oxide groups to adsorb dopamine. The oxidative etching further increases the peak current to 3.5 ± 0.2 times of the CFME, and ox-CNH/CFME had a limit of detection of 6 ± 2 nM. The dopamine anodic current at ox-CNH/CFME was stable for 8 h of continuous scanning. The ox-CNH/CFME enhanced the anodic peak current for other cationic neurotransmitters including epinephrine, norepinephrine, and serotonin, but less enhancement was found for ascorbic acid, showing higher selectivity for cationic molecules. CNHs also decreased tissue biofouling at CFME. Thus, electrodeposited CNHs are a promising new method for increasing the surface area and current of CFMEs for dopamine detection.