He succeeded Isidore Geoffroy Saint-Hilaire (1805-1861) as professor of zoology to the Faculty of Sciences at the University of Paris.
Gratiolet is remembered for his work in neuroanatomy, physiognomy and physical anthropology.
He did extensive research in the field of comparative anatomy, and performed important studies regarding the differences and similarities between human and various primate brains.
With Paul Broca (1824-1880) he performed correlative studies of aphasia and the frontal lobe.
Gratiolet was a vocal critic of Broca regarding the latter's belief that a larger brain equated to higher intelligence.