Nano-Sampling and Reporter Tools to Study Metabolic Regulation in Zebrafish

Abstract

In the past years, evidence has emerged that hallmarks of human metabolic disorders can be recapitulated in zebrafish using genetic, pharmacological or dietary interventions. An advantage of modeling metabolic diseases in zebrafish compared to other “lower organisms” is the presence of a vertebrate body plan providing the possibility to study the tissue-intrinsic processes preceding the loss of metabolic homeostasis. While the small size of zebrafish is advantageous in many aspects, it also has shortcomings such as the difficulty to obtain sufficient amounts for biochemical analyses in response to metabolic challenges. A workshop at the European Zebrafish Principal Investigator meeting in Trento, Italy, was dedicated to discuss the advantages and disadvantages of zebrafish to study metabolic disorders. This perspective article by the participants highlights strategies to achieve improved tissue-resolution for read-outs using “nano-sampling” approaches for metabolomics as well as live imaging of zebrafish expressing fluorescent reporter tools that inform on cellular or subcellular metabolic processes. We provide several examples, including the use of reporter tools to study the heterogeneity of pancreatic beta-cells within their tissue environment. While limitations exist, we believe that with the advent of new technologies and more labs developing methods that can be applied to minimal amounts of tissue or single cells, zebrafish will further increase their utility to study energy metabolism.

Differential Responses of Neural Retina Progenitor Populations to Chronic Hyperglycemia

“…However, our model provides a system to identify regenerative mechanisms that are robust to metabolic dysregulation. Another disadvantage relates to the small size of the zebrafish, which is advantageous for maintenance, but poses challenges for performing biochemical and metabolic analyses [71]. Whereas in rodents, glucose levels can be measured repeatedly in the same animals, our diabetic animals do not tolerate this.…”

Commensal Microbiota Regulate Vertebrate Innate Immunity-Insights From the Zebrafish

“…Fluorescence activated cell sorting (FACS) is one technological approach to overcome some of these limitations (31, 73). Additionally, the continued development of microdissection techniques using microneedles to sample small blood volumes or tissues will permit molecular analyse of distinct cellular populations from zebrafish larvae (130). These approaches, in combination with the advent of low-input molecular techniques (from PCR to single-cell sequencing), present exciting opportunities to parse the effects of the microbiota on rare and/or specific cell populations.…”

Genetically encoded thiol redox-sensors in the zebrafish model: lessons for embryonic development and regeneration

“…Isolating cells with specific redox signaling states for further studies could be accomplished using fluorescence activated cell sorting (FACS) based on sensor oxidation state, analogous to previous studies in zebrafish embryos targeting cells labelled according to signaling pathway activity, tissue identity or cell cycle phase (Dickmeis et al 2001;Dickmeis et al 2004;Wragg et al 2020). However, FACS may alter the redox state of cells (Llufrio et al 2018), so results should be compared with data obtained by laser capture microdissection (Veldman et al 2007;Zoidl et al 2004) or other nano-sampling techniques (Dickmeis et al 2019). Such single-cell based analyses should allow to produce a clearer picture of which H 2 O 2 effects in which tissues and cells are important for which developmental and regenerative processes.…”