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Scientific investigations, in general, and research in neuroscience, in particular, are becoming ever more complex and require the integration of different techniques. Behavioral assays, which are among the most frequently used methodologies in neuroscience, nowadays rely on advanced, sophisticated technologies that require proficient application. Therefore, behavioral core facilities are becoming essential support units, as they provide the specialized expert research services needed to conduct advanced neuroscience. We here review the lessons learned and insights gathered from managing behavioral core facilities in different academic research institutes. This review addresses several issues, including: the advantages of behavioral core facilities, considerations for establishing a behavioral core facility, and the methodological advances made through calibration and standardization of assay protocols and the development of new assays. Collectively, the review highlights the benefits of both working within and collaborating with behavioral core facility units and emphasizes the potential progress in neuro-phenotyping that such facilities provide.

Saltatorial locomotion is a type of hopping gait 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 strain of domesticated rabbits, the sauteur d’Alfort, that exhibits an abnormal locomotion behavior defined by the loss of the typical jumping that characterizes wild-type rabbits. Strikingly, individuals from this strain 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 populations of spinal cord interneurons. Our results show that RORB function is required for the performance of saltatorial locomotion in rabbits.

Author summary

Rabbits and hares have a characteristic jumping gait composed of an alternate rhythmical movement of the forelimbs and a synchronous bilateral movement of the hindlimbs. We have now characterized a recessive mutation present in a specific strain of domestic rabbits (sauteur d’Alfort) that disrupts the jumping gait. The mutation causing this defect in locomotion pattern occurs in the gene coding for the transcription factor RORB that is normally expressed in many regions of the nervous system especially in the spinal cord dorsal horn. Our results show that expression of RORB is drastically reduced in the spinal cord of affected rabbits which results in a developmental defect. This study is an advance in our understanding how locomotion is controlled in vertebrates.

Vertebrate locomotion is orchestrated by spinal interneurons making up a central pattern generator. Proper coordination of activity, both within and between segments, is required to generate the desired locomotor output. This coordination is altered during acceleration to ensure the correct recruitment of muscles for the chosen speed. The transcription factor Dmrt3 has been proposed to shape the patterned output at different gaits in horses and mice. Here, we characterized dmrt3a mutant zebrafish, which showed a strong, transient, locomotor phenotype in developing larvae. During beat-and-glide swimming, mutant larvae showed fewer and shorter movements with decreased velocity and acceleration. Developmental compensation likely occurs as the analyzed behaviors did not differ from wild-type at older larval stages. However, analysis of maximum swim speed in juveniles suggests that some defects persist within the mature locomotor network of dmrt3a mutants. Our results reveal the pivotal role Dmrt3 neurons play in shaping the patterned output during acceleration in vertebrates.

Skilled forelimb movements are initiated by feedforward motor commands conveyed by supraspinal motor pathways. The accuracy of reaching and grasping relies on internal feedback pathways that update ongoing motor commands. In mice lacking the axon guidance molecule EphA4, axonal misrouting of the corticospinal tract and spinal interneurons is manifested, leading to a hopping gait in hindlimbs. Moreover, mice with a conditional forebrain deletion of EphA4, display forelimb hopping in adaptive locomotion and exploratory reaching movements. However, it remains unclear how loss of EphA4 signaling disrupts function of forelimb motor circuit and skilled reaching and grasping movements. Here we investigated how neural circuits controlling skilled reaching were affected by the loss of EphA4. Both male and female C57BL/6 wild-type, heterozygous EphA41/2, and homozygous EphA42/2 mice were used in behavioral and in vivo electrophysiological investigations. We found that EphA4 knock-out (2/2) mice displayed impaired goal-directed reaching movements. In vivo intracellular recordings from forelimb motor neurons demonstrated increased corticoreticulospinal excitation, decreased direct reticulospinal excitation, and reduced direct propriospinal excitation in EphA4 knock-out mice. Cerebellar surface recordings showed a functional perturbation of the lateral reticular nucleus-cerebellum internal feedback pathway in EphA4 knock-out mice. Together, our findings provide in vivo evidence at the circuit level that loss of EphA4 disrupts the function of both feedforward and feedback motor pathways, resulting in deficits in skilled reaching.

Nicotine is a commonly used drug that has been extensively studied for decades. Some of these studies have found cognitive enhancing effects; however, they were usually based on moderate and not the higher doses to which the regular consumer might be accustomed. Here we investigate how higher doses of nicotine may influence the performance of mice in object recognition and in the Y-maze. Further, we examined specific circuits underlying the hippocampal-dependent effects by targeting a subgroup of inhibitory interneurons, referred to as OLMα2 cells. We subjected mice to nicotine during an object recognition task and a working memory task. We found that a high dose of 1.5 mg/kg nicotine impaired memory performance in the object recognition task but not in the working memory task. It was previously demonstrated that OLMα2 cells bidirectionally affect learning in the object recognition task and that these cells respond to nicotine. Subsequently, we subjected mice to nicotine while optogenetically inhibiting OLMα2 cells and found that this intervention reversed the nicotine-induced memory impairment. We conclude that some learning and memory effects from nicotine are hippocampus dependent and are probably mediated by OLMα2 cells.

It is becoming increasingly clear that the hippocampus is functionally diverse across its longitudinal axis. The ventral hippocampus is known to participate anxiety-related behaviors and may, together with the basolateral amygdala (BLA), facilitate threat aversion. Nevertheless, studies of mechanisms and circuit organization for processing value related cues are scarce. Here we investigate a microcircuit involving a subgroup of interneurons, referred to as Oriens lacunosum-moleculare cells, defined by their expression of the nicotinic receptor alpha2 subunit (OLMα2). Such cells can bidirectionally affect the response to predator odor as well as the encoding of object memories. In tracing experiments, we found that the basolateral amygdala mainly projects to the ventral hippocampus, whereas the medial amygdala and claustrum projects to the intermediate hippocampus. Moreover, we found that BLA inputs inhibit intrinsic hippocampal slow oscillation.  Optogenetic stimulation of OLMα2 cells in the intermediate hippocampus caused an increase in approach of objects. Inhibition of ventrally located OLMα2 cells cause an increased avoidance to natural aversive stimuli and could entrain aversion to value neutral odors. In contrast, theta II oscillations, which predominantly appear in the ventral hippocampus during anxiety related behaviors, were absent during object recognition. Theta II oscillations only appeared when the animal was naïve to the arena setting. We conclude that there are functional differences between the intermediate and ventral hippocampus and that OLMα2 cells, in addition to sensory inputs, process emotional value signals from the amygdala.