Measuring behavioral and endocrine responses to novelty stress in adult zebrafish

Abstract

Several behavioral assays are currently used for high-throughput neurophenotyping and screening of genetic mutations and psychotropic drugs in zebrafish (Danio rerio). In this protocol, we describe a battery of two assays to characterize anxiety-related behavioral and endocrine phenotypes in adult zebrafish. Here, we detail how to use the 'novel tank' test to assess behavioral indices of anxiety (including reduced exploration, increased freezing behavior and erratic movement), which are quantifiable using manual registration and computer-aided video-tracking analyses. In addition, we describe how to analyze whole-body zebrafish cortisol concentrations that correspond to their behavior in the novel tank test. This protocol is an easy, inexpensive and effective alternative to other methods of measuring stress responses in zebrafish, thus enabling the rapid acquisition and analysis of large amounts of data. As will be shown here, fish anxiety-like behavior can be either attenuated or exaggerated depending on stress or drug exposure, with cortisol levels generally expected to parallel anxiety behaviors. This protocol can be completed over the course of 2 d, with a variable testing duration depending on the number of fish used.

Effect of novelty stress on behavioral responses of Danio rerio and assessment of dose-dependent effects of anxiolytics of benzodiazepine structure with phenazepam as an example

“…Калуев и др. [9] применили показатели тревожности и адаптирова-ли их непосредственно для Danio rerio. Рыбка по-мещается сначала в мерный стакан с растворенным фармакологическим веществом (или водой), а за-тем в небольшой просмотровый аквариум, где ре-гистрируются двигательная активность, фризинг, число переходов в верхнюю половину аквариу ма и время нахождения в последней.…”

Habenular kisspeptin modulates fear in the zebrafish

“…To evaluate the effect of Kiss1 on anxiety, a novel tank diving test was performed (n = 20 per group) according to the protocol implemented by Cachat and coworkers (14). In brief, the following behavioral parameters were recorded: total time spent at the bottom-half of the tank, number of top-to-bottom transitions, and total distance traveled in either the top or bottom half of the tank.…”

“…In this study, to systematically examine anxiety and fear, we used two different experimental procedures. The novel tank diving test was used to analyze anxiety (bottom dwelling and top-to-bottom transitions in the tank) (14), and alarm substance (also known as alarm pheromone) was introduced to create fear response (erratic movements and freezing) (15). Further, to substantiate the role of endogenous Kiss1 in the absence of fish-specific Kiss-R1 antagonist and zebrafish mutants (or knockout) of kiss1/kissr1, we conjugated zebrafish Kiss1 peptide to saporin (SAP), a ribosomeinactivating protein (16), to selectively inactivate Kiss-R1-expressing neurons, and we examined AS-evoked fear response.…”

Automated High-throughput Behavioral Analyses in Zebrafish Larvae

“…Zebrafish larvae are ideally suited for high-throughput analyses because of their small size 7,8 . Software is commercially available for automated analyses of behavior in larval and adult zebrafish [9][10][11][12][13][14] . However, this software can be expensive and has limited options for measuring complex behaviors of zebrafish larvae in multi-well plates.…”