Giuseppe Crescimanno

The Hole-Board is an ethologically based tool for investigating the anxiety-related behavior of rats following manipulation of the central anxiety level. The present paper aims to assess behavioral patterning following pharmacological manipulation of emotional assets in Wistar rats tested in this experimental apparatus. For this purpose, the behavior of three groups of rats injected with saline, diazepam or FG7142 was evaluated using conventional quantitative and multivariate T-pattern analyses. The results demonstrate that quantitative analyses of individual components of the behavior, disjointed from the comprehensive behavioral structure, are of narrow utility in the understanding of the subject’s emotional condition. Among the components of the behavioral repertoire in rodents tested in the Hole-Board, Edge-Sniff and Head-Dip represent the most significant ones to rate anxiety level. They are characterized by a strong bivariate relationship and are also firmly part of the behavioral architecture, as revealed by the T-pattern analysis (TPA), a multivariate technique able to detect significant relationships among behavioral events over time. Edge-Sniff → Head-Dip sequences, in particular, are greatly influenced by the level of anxiety: barely detectable in control animals, they completely disappear in subjects with a reduced level of anxiety and are present in almost 25% of the total of T-patterns detected in subjects whose anxiety level increased.

Temporal pattern analysis is an advanced multivariate technique able to investigate the structure of behavior by unveiling the existence of statistically significant constraints among the interval length separating events in sequence. If on the one hand, such an approach allows investigating the behavioral response to pain in its most intimate and inner features, on the other hand, due to the meaning of the studies on pain, it is of relevant importance that the results utilize intuitive and easily comprehensible ways of representation. The aim of this paper is to show various procedures useful to represent the results originating from the multivariate T-pattern analysis of the behavioral response to pain in Wistar rats tested in a hot-plate and IP injected morphine or saline as a control.

Rationale Little is known about the structural characteristics of the behavior of rats with enhanced anxiety level. To fill this gap, a study was undertaken where effects of an anxiogenic drug were examined on behavioral structure of rats tested in hole board. Objectives This study investigates effects of increased anxiety level on the structure of the behavior of rats tested in hole board Methods Different doses of FG7142 (1, 4, 8 mg/kg IP), a potent anxiety-inducing drug, were administered to three groups of male Wistar rats. A further group was administered saline. Experiments were recorded through a digital camera. Quantitative and multivariate approaches were applied. Results Percent distributions and durations showed increases of immobile sniffing, rearing, head dip, and edge sniff and a significant reduction of grooming activities and of walking. In addition, a decrease of head dip/edge sniff ratio was detected. Transition matrices evidenced that FG7142 provoked evident modifications of behavioral structure mainly of general exploration of environment and focused exploration of the hole. Finally, adjusted residuals showed a reduced effectiveness of FG7142 on transitions from head dip to edge sniff; on the contrary, transitions from edge sniff to head dip underwent evident dose-dependent changes. Conclusions Present study provides a useful tool to analyze behavioral responses to different anxiety conditions. Accordingly, it is demonstrated that a condition of increased anxiety deeply modifies the structure of male Wistar rat’s behavior in hole board. In addition, our results suggest that evaluation of head dip/edge sniff ratio can be considered a reliable index to appraise effects of pharmacological manipulation of anxiety and related behavioral elements.