Extracellular ATP and other extracellular nucleotides acting via P2 receptors in the inner ear initiate a wide variety of signalling pathways important for regulation of hearing and balance. Ectonucleotidases are extracellular nucleotide-metabolising enzymes that modulate purinergic signalling in most tissues. Major ectonucleotidases in the cochlea are likely members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family. In this study, we provide a detailed description of NTPDase1 and NTPDase2 distribution in cochlear tissues using immunocytochemistry. E-NTPDase immunoreactivity was not equally distributed in the tissues bordering scala media. It was observed in the organ of Corti, including sensory and supporting cells, but was notably absent from Reissner's membrane and most of the marginal cells of the stria vascularis. NTPDase1 expression was most prominent in the cochlear vasculature and cell bodies of the spiral ganglion neurones, whereas considerable NTPDase2 immunoreactivity was detected in the stria vascularis. Both E-NTPDases were expressed in the cuticular plates of the sensory hair cells and nerve fibres projecting from the synaptic area underneath the inner and outer hair cells. E-NTPDase localisation corresponds to the reported distribution of some P2X receptor subunits (P2X(2) in particular) in sensory, supporting and neural cells and also P2Y receptor distribution in the vasculature and secretory tissues of the lateral wall. The role for E-NTPDases in purinergic signalling is most likely to regulate extracellular nucleoside triphosphate and diphosphate levels and thus provide termination for extracellular ATP signalling that has been linked to control of cochlear blood flow, electrochemical regulation of sound transduction and to neurotransmission in the cochlea.