Impact of post-hatching maturation on the pharmacokinetics of paracetamol in zebrafish larvae

Abstract

Zebrafish larvae are increasingly used in pharmacological and toxicological studies, but it is often overlooked that internal exposure to exogenous compounds, rather than the incubation medium concentration, is driving observed effects. Moreover, as the zebrafish larva is a developing organism, continuous physiological changes impact pharmacokinetic or toxicokinetic processes like the absorption and elimination of exogenous compounds, influencing the interpretation of observations and conclusions drawn from experiments at different larval ages. Here, using paracetamol as paradigm compound, mathematical modelling is used to quantify absorption and elimination rates from internal exposure over time profiles after waterborne treatment, as well as changes in these parameters in post-hatching larvae of 3, 4, and 5 days post fertilisation (dpf). An increase of 106% in absorption rate was observed between 3 and 4 dpf, but no further increase at 5 dpf, and an increase of 17.5% in elimination rate for each dpf. Paracetamol clearance, determined from elimination rate constants and reported total larval volumes of 253, 263, and 300 nL at 3, 4, and 5 dpf respectively, correlates best with higher vertebrates at 5 dpf. This suggests that when studying direct effects of exogenous compounds, experiments with zebrafish larvae are best performed at 5 dpf.

Effective processing and evaluation of chemical imaging data with respect to morphological features of the zebrafish embryo

“…Contrary to the bromine case, the 4-iodophenol showed a more dynamic behavior: With continuing exposure and ZFE development, the iodine is transferred from the yolk to the GI tract. This may be explained by an increased metabolizing capacity of older ZFEs [ 42 , 43 ] as well as the development of the elimination pathways via the liver and kidney [ 44 ]. It has recently been shown that the distribution of toxicants between the yolk and the ZFE body of developing ZFEs can be highly dynamic [ 40 ].…”

Anti‐tuberculosis effect of isoniazid scales accurately from zebrafish to humans

“…Quantification of internal drug exposure in zebrafish larvae after external, waterborne drug treatment is essential for reliable interpretation of drug response, but because of small sample volumes and low concentrations, this is not straightforward. We have built upon recent work to develop an ultra‐sensitive LC–MS based quantification method (Kantae et al, 2016; Van Wijk, Krekels, Kantae, Harms, et al, 2019; Van Wijk, Krekels, Kantae, Ordas, et al, 2019). This methodology proved sensitive enough to quantify isoniazid in larval homogenates and blood samples of only nanolitres in volume.…”

Mechanistic and Quantitative Understanding of Pharmacokinetics in Zebrafish Larvae through Nanoscale Blood Sampling and Metabolite Modeling of Paracetamol

“…In the case of our paradigm compound paracetamol (acetaminophen), internal paracetamol amounts were subsequently used to quantify the pharmacokinetic processes of absorption and elimination through quantification of the rate of absorption and a clearance relative to total larval volume. Although the clearance derived from relative values in zebrafish larvae scaled quite reasonably to reported absolute clearance values in higher vertebrates ((Kantae et al, 2016;Van Wijk et al, 2019)), accurate scaling of clearance requires absolute clearance values in zebrafish larvae. However, quantification of absolute clearance requires blood concentrations, which will also allow for the estimation of a distribution volume, but blood-sampling methods for zebrafish larvae that are only a few hundred nanoliters in total volume (Guo et al, 2017) have not yet been described.…”