The ink gland in cephalopods is a modified part of the digestive tract that generates melanin used in defense activities against predators. In the present study, we document the histological structure of the ink gland of the cuttlefish Sepia pharaonis following light microscopy (LM) and transmission electron microscopy (TEM) examinations. Based on the observations, we infer the series of cellular events leading to the formation and secretion of melanin. LM observations revealed that the cuttlefish ink gland was cord-like, and could be separated into two distinct regions with different functions, based on markedly different cellular and biochemical characteristics. The region with the function of secreting ink was the epithelial cell region (i.e., mature ink gland cells) and was mainly composed of melanosomes, rough endoplasmic reticulum, Golgi apparatus, and mitochondria. Conversely, the connective tissue region was the site of formation of cells and energy supply. In addition, the most probable sequence of activities from melanogenesis to the release, based on TEM observations, is as follows: (a) formation of a matrix of opaque electronic substances (melanin precursors); (b) melanization in the precursor matrix to form melanin-like particles; (c) further melanin granule development and enlargement, and gradual intensifying of the hue; (d) production of melanin granules (melanosomes) in the precursor matrix; (e) binding of melanosomes to the cytoskeleton and migration to the cell surface; (f) fusion of melanosomes with the cell membrane at the apical pole, and discharge of melanin granules into intercellular spaces or lumen by exocytosis or cell fragmentation. We also investigated how the process of continuous ink release influences ink gland histology and ultrastructure. Inking stimulation induced the release of large amounts of ink by epithelial cells, and was associated with cell disorganization, and severe cellular vacuolization, in addition to severe organelle damage.