Autophagy is a major intracellular pathway for the degradation and recycling of long-lived proteins and cytoplasmic organelles. Like apoptotic programmed cell death, autophagy is an essential part of growth regulation and maintenance of homeostasis in multicellular organisms. Autophagic vacuole formation is also activated as an adaptive response to a variety of extracellular and intracellular stimuli, including nutrient deprivation, hormonal or therapeutic treatment, bacterial infection, aggregated and misfolded proteins and damaged organelles. Mediators of class I and class III PI3 kinase signaling pathways and trimeric G proteins play major roles in regulating autophagosome formation during the stress response. Defective autophagy is the underlying cause of a number of pathological conditions, including vacuolar myopathies, neurodegenerative diseases, liver disease, and some forms of cancer. This chapter provides an overview of the morphology and molecular basis of autophagosome formation and offers a glimpse into the role of autophagy in normal growth and development, while discussing the pathological implications of its deregulation.