Parisian-born Charcot studied medicine in his native city and received his MD in 1853. His interest in disease of the nervous system led to his appointment, in 1862, to the Salpêtrière Hospital for nervous and mental disorders. This marked the beginning of a long and distinguished association. Charcot described the pathological changes associated with several degenerative conditions of the nervous system, including the disintegration of ligaments and joint surfaces (known as Charcot's disease) that occurs in advanced stages of locomotor ataxia. His studies of brain damage in cases of speech loss (aphasia) and epilepsy supported the findings of his contemporary, Paul Broca, that is, different bodily functions are controlled by different regions of the cerebral cortex.

In 1872, Charcot was appointed professor of pathological anatomy at the faculty of medicine and later (1882) became professor of neurology at the Salpêtrière. He was increasingly concerned with the link between mind and body in cases of hysteria and trauma. With his eloquent manner and a dramatic presentation of his lectures on a small stage, he became a widely celebrated teacher. Among many famous students was Sigmund Freud, who was influenced by Charcot's use of hypnosis on patients.

Charcot's son, Jean, became a famous polar explorer.