Neurobiology of pair bonding in Chaetodon lunulatus (f. Chaetodontidae), with respect to isotocin, arginine vasotocin V1a, dopamine D1 and D2, and mu-opioid receptor systems

This collection contains data pertaining to the neurobiology of pair bonding in Chaetodon lunulatus butterflyfish, with respect to isotocin, arginine vasotocin V1a, dopamine D1 and D2, and mu-opioid receptor systems.Two data sets are included in the collection: 1) effects of pharmacological administration of saline, and receptor blockades on baseline preferential affiliation with partner (partner preference) in males. 2) brain region specific gene expression (mRNA abundance) of receptors in pair bonded and solitary individuals.Summary of methods for effects of pharmacological administration of receptor blockades on partner preference (dataset 1): Receptor types examined include isotocin receptor (ITR), arginine vasotocin V1a receptor (V1aR), dopamine D1 subtype receptor (D1R), and mu-opioid receptor (MOR). Study was conducted on adult males. Baseline preferential affiliation with partner over non-partner (“partner preference”) was established using a partner preference testing paradigm (see description and methods of partner preference test in “Nowicki: Classification of social systems of six butterflyfish species (f. Chaetodontidae; g. Chaetodon) at Lizard Island, Great Barrier Reef, Australia, 2013-2015”) data collection. Following, the focal male was removed and delivered either a saline control or receptor blockade treatment (ITR blockade: desGly-NH2-d(CH2)5[D-Try2,Thr4]OVT; V1aR blockade: SR49059; D1R blockade: SCH23390; MOR blockade: CTAP) via intramuscular injection. The positions of intra- and extra-pair females were then switched and opaque barriers were placed between chambers, and the treatment male was re-introduced into the central zone of the center chamber to undergo drug activation (or in the case of saline a “sham” drug activation) in isolation for 30 min. Thereafter, the opaque barriers were removed, and tests of partner preference were repeated. Each male was only tested once.Summary of methods for comparing brain region specific gene expression (mRNA abundance) of receptors between pair bonded and solitary individuals (dataset 2): Paired and solitary individuals were collected from the wild, sacrificed, sexed histologically, and brains dissected and frozen in liquid nitrogen. Frozen brains were sectioned coronally, brain regions of interest were removed using a hand-held tissue extraction tool and butterflyfish brain atlas as reference, RNA was extracted and reverse transcribed into cDNA. Using C. lunulatus transcriptome as a reference, primers were designed for pre-amplifying target genes using PCR, and quantified target genes using RT qPCR. Target genes included: ITR, V1aR, D1R, D2R, MOR, and normalizer gene 18S ribosomal. During qPCR, samples were run in technical triplicate, from which gene expression (Ct) values was averaged. Since qPCR assay efficiency was not the same across individual assays, averaged gene expression (Ct) values were standardized to assay efficiency prior to normalizing to 18S ribosomal RNA.This project was conducted in accord to permits and animal ethic standards put forth by James Cook University and granted to JPN.