Establishment of open-source semi-automated behavioral analysis system and quantification of the difference of sexual motivation between laboratory and wild strains

Abstract

Behavioral analysis plays an important role in wide variety of biological studies, but behavioral recordings often tend to be laborious and are associated with inevitable human-errors. It also takes much time to perform manual behavioral analyses while replaying the videos. On the other hand, presently available automated recording/analysis systems are often specialized for certain types of behavior of specific animals. Here, we established an open-source behavioral recording system using Raspberry Pi, which automatically performs video-recording and systematic file-sorting, and the behavioral recording can be performed more efficiently, without unintentional human operational errors. We also developed an Excel macro that enables us to easily perform behavioral annotation with simple manipulation. Thus, we succeeded in developing an analysis suite that mitigates human tasks and thus reduces human errors. By using this suite, we analyzed the sexual behavior of a laboratory and a wild medaka strain and found a difference in sexual motivation presumably resulting from domestication.

Sex differences in autism‐like behavior and dopaminergic neurons in substantia nigra of juvenile mice prenatally exposed to valproate

“…In male mice, our data indicate that VPA modifies the number of TH + neurons in the substantia nigra on the PND 31. This effect might be accomplished by a reduction in the number of TH + precursor cells that appear in the midbrain since embryonic development (E12.5) (Blaess et al, 2011;Sonne et al, 2022) and are significantly vulnerable to VPA exposure (Roullet et al, 2013). This vulnerability to VPA may explain some differences found in structural changes described in early life (PND 7) with other VPA models (Ádám et al, 2020).…”

“…Then, we quantified the absolute number of marbles buried and the frequency of digging behavior (Figure 2b). The Ethogramer is a semi‐automated tool (Tomihara et al., 2021) that was used to record the time and frequency of behaviors and design a visual raster of the recorded behaviors (Figures 1c and 2c). The Ethogramer assigns a color bar that identifies a specific behavior, and the width of the bar indicates the time spent on that behavior.…”

Involvement of the kisspeptin system in regulation of sexual behaviors in medaka

“…Medaka is a small teleost that has many experimental advantages because its genome database is almost fully available for various genetic manipulations. It also shows stereotypical sequence of sexual behaviors that is necessary for successful spawning 33 , 34 , 35 and regularly spawns every morning under appropriate breeding conditions. In addition, unlike mammals, kisspeptin in teleosts is not involved in the HPG axis regulation, 6 indicating that medaka is an advantageous model animal to search for novel and reproductive state-dependent functions such as sexual behavior in vertebrates by using a combination of genome-editing and behavior analysis.…”

A novel open-source raspberry Pi-based behavioral testing in zebrafish

“…Nowadays, animal behavior are usually observed by expensive commercial devices or human eyes, which are time-and labor-intensive, defying high-throughput detection. In addition, video capturing of animal behavior and subsequent computer analysis can automatically obtain and process a large number of video signals to give accurate description of behavior phenotype, allowing high-throughput study of animal behavior changes [7][8][9]. However, most of the zebrafish behavior assay setup on the market is sophisticated and expensive [10][11][12][13], thus greatly limited its application.…”

“…It has been widely used in laboratories, education, robots, and other fields [14][15][16][17]. In addition, Raspberry Pi's rich remoteoperated features can greatly reduce experimental bias and error, thus easy for automation to obtain huge amount of accurate dataset [9]. These advantages make Raspberry Pi an appealing choice to develop low-cost, and high-throughput animal behavior assay setup.…”