26understand the mechanisms underlying parental behavior and its evolution are comparative 51 studies across closely-related species that vary in parental care strategies. 52Parental care can be conceptualized as a complex set of inter-related behaviors controlled 53 by brain regions involved in the integration of sensory, social, motivational, and cognitive aspects 54 of care [6]. Across vertebrates, these functions are largely performed by the social decision-55 making network (SDMN; [7]), a highly interconnected group of evolutionarily ancient and 56 functionally conserved brain regions. Although studies on the neural mechanisms of parental 57 behavior are sparse outside mammals, and particularly lacking in amphibians and reptiles, the 58 SDMN provides an ideal starting point for this work as network nodes and connectivity are well 59 understood, highly conserved, and behaviorally important ligand/receptor complexes have been 60 extensively studied. 61Dendrobatid poison frogs show remarkable diversity in parental care across closely 62 related species, including male uniparental care, female uniparental care, and biparental care. 63Parental care in poison frogs involves egg attendance during embryo development, generally 64 followed by transportation of tadpoles "piggyback" to pools of water upon hatching [8][9][10]. In some 65 species, mothers regularly return to nourish growing tadpoles with unfertilized, trophic eggs until 66 the laboratory, allowing us to identify both parental individuals and their non-caregiving partners. 87To control for effects of experience, all pairs successfully reared at least one clutch from egg-88 laying through tadpole transport prior to the experiment. For the non-parental group, we collected 89 frog pairs between parental bouts when they were not caring for eggs or tadpoles, collecting 90 individuals of both the caregiving sex (non-transport; n=10 D. tinctorius, n=7 R. imitator) and their 91 opposite sex partners (non-transport partner; n=9 D. tinctorius, n=8 R. imitator). For the tadpole 92 5 transport group, when we found transporting frogs, we collected both the tadpole transporting 93 individual (tadpole transporter; n=13 D. tinctorius, n=7 R. imitator) and its opposite sex, non-94 transporting partner (transport partner; n=11 D. tinctorius, n=6 R. imitator). All brain tissue was 95 collected in an identical manner: frogs were captured, anesthetized with benzocaine gel, weighed 96 and measured, and euthanized by rapid decapitation. This entire process took less than 5 97 minutes. All procedures were approved by the Harvard University Animal Care and Use 98 Committee (protocol no. 12-10-1). 99 100
Field sample collection 101Oophaga sylvatica (Puerto Quito-Santo Domingo population) were collected in field 102 enclosures in Ecuador in April and May of 2016. We collected non-parental control females (N=8) 103 from enclosures containing only mature females to ensure that frogs were not currently caring for 104 eggs or tadpoles. We collected tadpole transporting females (N=5) fr...