The mouse gubernaculum undergoes inguinoscrotal migration in the first postnatal week and also shows rhythmic contractions in organ culture in response to calcitonin gene-related peptide (CGRP). This study aimed to investigate the ontogeny of gubernacular contractile activity and effect of CGRP in organ culture in relation to the normal time of migration. Two hundred eighty gubernacula from male mice of 17 and 19 days' gestation, and of postnatal days 0,2,4,7, 10, and 14 were studied. Half were cultured with CGRP 714 nmol/L, and the other half were cultured without CGRP as controls. All were examined daily (for 7 days) for contractions. In the control group, the cumulative percentages of contractile gubernacula increased from 5% to 100% with age; in the CGRP group, these values ranged from 65% to 100%. There were significant differences between the two groups from prenatal day 17 to postnatal day 4 (P values ranged from less than .001 to less than 0.05, respectively). With increasing age in the CGRP group, the highest contractile rates were observed at the fewest number of days in culture. From prenatal day 17 to postnatal day 4, the gubernaculum had increasing endogenous contractility, but there was low endogenous contractility without exogenous CGRP. Contractility of gubernacula was enhanced strongly by exogenous CGRP, and the culture days of the peak contractile rates lined up in the reverse order. These results suggest that the days of optimal gubernacular contraction with CGRP in vitro are in keeping with the days of natural migration in vivo. This is consistent with the hypothesis that CGRP (released from the genitofemoral nerve) is important for gubernacular migration.