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Their localization in the pituitary during gonadal sex differentiation has been studied mainly in southern (<jats:italic>Oryzias latipes<\/jats:italic>) strains of medaka fish, with that in northern medaka (<jats:italic>O. sakaizumii<\/jats:italic>) remaining poorly understood. Hence, in this study, we characterized gonadal differentiation and gonadotrophic cells during sex differentiation in two northern strains (HNI and Kaga) and two southern strains (Hd\u2010rR and d\u2010rR\/Tokyo). All strains exhibited similar sex differentiation at hatching, such as (1) sex difference in germ cell number (XX\u2009&gt;\u2009XY), and (2) the transition of XX germ cells into meiosis, and (3) presence of glycoprotein\u2010\u03b1 (Gpa)\u2010 positive cells. However, follicle\u2010stimulating hormone\u2010\u03b2 (Fshb)\u2010positive cells were first detected in the pituitary 1 day post\u2010hatching in HNI. Exposure to high\u2010temperature conditions and to cortisol in a dose dependent manner resulted in the localization of Fshb cells in the pituitary at hatching. This study demonstrates differences in gonadotropin subunit expression between northern and southern strains of medaka, and suggests that Fsh is not involved in early gonadal sex differentiation, such as the sex difference in germ cell number, and that high\u2010temperature induce Fshb expression via cortisol production in medaka. <jats:italic>J. Exp. 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