Colonisation of freshwater habitats by marine animals is a remarkable
evolutionary event that has enriched biodiversity in freshwater
ecosystems. For successful freshwater colonisation, high physiological
plasticity is presumed to be necessary, but its evolutionary basis has
not been detailed. Marine-originated amphidromous species, which
regularly migrate between freshwater and marine environments, have
repeatedly lost migratory behaviour in many lineages, which sometimes
triggered species radiation in freshwater habitats. Since amphidromous
species typically visit the sea during the larval period, the difficulty
in the evolution of larval freshwater tolerance is a bottleneck for
freshwater colonisation. To elucidate the key evolutionary changes that
enhance the physiological plasticity for freshwater colonisation, we
compared larval gene expression changes depending on salinity conditions
among three congeneric amphidromous goby species (Gymnogobius) with
varying dependences on freshwater habitats. First, an otolith
microchemical analysis and rearing experiment under laboratory
conditions confirmed the presence of freshwater residents only in G.
urotaenia and higher larval survivorship of this species both in
seawater and freshwater conditions than the obligate amphidromous G.
petschiliensis and G. opperiens. Larval whole-body transcriptome
analysis revealed that G. urotaenia exhibited the greatest differences
in the expression levels of several osmoregulatory genes, including
aqp3, which is critical for water discharge from their body during early
fish development. Thus, we obtained the results that consistently
support the importance of enhanced osmoregulatory plasticity for
establishing freshwater forms, and further identified some important
evolutionary changes for larval freshwater adaptation and colonisation
in the goby group.