Following yolk-sac absorption and gravel emergence pink salmon (Oncorhynchus gorbuscha) migrate into seawater (SW) at as small as 0.2 g. This life-history strategy is in contrast with most anadromous salmonid species that generally spend 1-2 years growing in fresh water (FW) and physiologically preparing for life in SW before they migrate to SW as smolts. This study characterized for the first time the ontogeny of SW tolerance in pink salmon around the period of yolk-sac absorption. Post-hatch juvenile pink salmon were either held in FW for 26 weeks or transferred to SW every two weeks for 20 weeks to follow % survival, whole body (WB) Na+ and water content, as well as changes in wet and dry mass, gill Na+K+ATPase (NKA) activity and α1a and α1b mRNA isoform expression. An increase in gill NKA activity and the ratio of the α1b/α1a isoform expression, a plateau in WB water and Na+ levels, and the switch from catabolic to anabolic growth were all observed at the time of yolk-sac absorption in fish retained in freshwater. At this time, morbidity following subsequent SW transfer fell to 0% from a high of 100% for newly hatched alevins, but then rose to 25% in older fry, suggesting that a window of increased salinity tolerance exists for pink salmon at the time of yolk-sac absorption. This proposed window of SW tolerance is similar to the smolt window that has been identified for other salmonids; but in pink salmon appears to be endogenously mediated, as fish were reared under constant (12L:12D) photoperiod and at 5˚C throughout the study. Moreover, smoltification is incomplete since transfer to SW further elevated gill NKA activity and increased gill NKA α1b/α1α isoform expression ratio 8-fold at yolk-sac absorption. Thus, even the most SW-tolerant fish were not fully prepared for SW before entry, but responded directly to SW by further increasing hypo-osmoregulatory ability. This study filled the previously existing void of knowledge regarding the acquisition of salinity tolerance in juvenile pink salmon.