Mesodiencephalic dopamine neurons play a central role in the regulation of a wide range of brain functions ranging from voluntary movement to reward associated behaviours. These functions are regulated by distinct subtypes of dopamine neurons located in the ventral midbrain substantia nigra pars compacta and ventral tegmental area that project to different brain regions by forming the nigrostriatal, mesolimbic, and mesocortical pathways. The molecular mechanisms that drive the midbrain dopaminergic trajectory formation are largely unknown. Autophagy is the major cytoplasmatic turnover pathway and has been shown to be important to neural system development. Here we show that autophagy machinery is present in the growth cones of dopaminergic neurons and is temporally regulated during their growth and guidance. Furthermore, autophagy level changes dynamically in dopaminergic neurons in response to both chemo-repulsive and chemo-attractive guidance cues. To characterize the role of autophagy in dopaminergic axon growth/guidance, we used CRISPR-Cas9 gene editing as well as a conditional knock-out mice (cKO) for the essential autophagy genes (Atg12, Atg5) deleted in dopaminergic neurons. ATG5 cKO axons exhibit axonal swellings and decreased branching in vitro and in vivo, likely due to aberrant microtubule looping. Strikingly, deletion of autophagy-related genes blunted completely the response of dopaminergic neurons to chemo-repulsive and chemo-attractive guidance cues. Our data demonstrate that autophagy plays a central role to tightly regulate dopaminergic neurons development and improve our understanding about basic physiological processes orchestrating axonal growth and guidance.