The Harderian gland in domestic birds is a major paraocular excretory gland that has an important role in tear production, as well as in the immune protection of the conjunctiva surface. The exocrine and lymphatic tissue of this gland has been described, but information about nerve supply is still very scarce in the present literature, so the main aim of this study was to provide more detailed and precise data about it. The research was conducted on 20 laying hens (Lohmann Brown) aged 4 months. Dissection and histomorphological examination show that the gland is innervated by the branch of the III (n. oculomotorius), V (n. trigeminus), and VII (n. facialis) cranial nerve. Before reaching the gland, the branches of these nerves enter into the ganglion ethmoideum, and later from this emerge several, thin postganglionic nerve fibers (ramus glanduale membrane nictitanits) that penetrate the capsule of the gland at its anterior extremity. The present study describes in detail the innervation of the Harderian gland, and this data will be useful for further research.