Retinoid-Related Orphan Nuclear Receptor RORB is Required for Saltatorial Locomotion in Rabbits

Saltatorial locomotion is a type of gait based on hopping that in mammals can be found in rabbits, hares, kangaroos, and some species of rodents. The molecular mechanisms that control and fine tune the formation of this type of gait are unknown. Here, we take advantage of one breed of domesticated rabbits, the sauteur d’Alfort, that exhibits an abnormal locomotion behavior characterized by the loss of the typical jumping that characterizes wild-type rabbits. Strikingly, individuals from this breed frequently adopt a bipedal gait using their front legs. Using a combination of experimental crosses and whole genome sequencing, we show that a single locus containing the RAR related orphan receptor B gene (RORB) explains the atypical gait of these rabbits. We found that a splice-site mutation in an evolutionary conserved site of RORB results in several aberrant transcript isoforms incorporating intronic sequence. This mutation leads to a drastic reduction of RORB-positive neurons in the spinal cord, as well as defects in differentiation of the DMRT3-expressing population of interneurons. Our results demonstrate that RORB plays an important role in neuronal differentiation of the rabbit spinal cord that is required for normal saltatorial locomotion. They further suggest that the DMRT3 gene appears to have a general role in regulating gait across species and different types of locomotion.