The zebrafishgoosepimples/myosin Vbmutant exhibits cellular attributes of human microvillus inclusion disease

Abstract

Bullet points (Key findings):• myosin Vb is expressed in the zebrafish intestine.• goosepimples/myosin Vb function is essential for the morphogenesis, cell shape modulation, secretion of glycoproteins and prevention of the formation of microvillus inclusions in the zebrafish intestine.• The function of myosin Vb in the intestine is conserved between fish and humans.• The goosepimples mutant is a good animal model for microvillus inclusion disease in humans. Grant Sponsor: Wellcome Trust-DBT India alliance (MS; 500129-Z-09-Z), Department of Science and Technology, India (MS; SR/S2/RJN-06/2006), TIFR-DAE (MS; 12P-121). 2 Abstract Background: Microvillus inclusion disease (MVID) is a life threatening enteropathy characterised by perpetual diarrhoea. Recent analysis has revealed that enterocytes in MVID patients exhibit reduction of microvilli, presence of microvillus inclusion bodies and intestinal villus atrophy. Genetic linkage analysis has identified mutations in myosin Vb gene as the main cause of MVID. An animal model that develops ex-utero and is tractable genetically as well as by microscopy would be highly useful to study the cellular basis of the formation of inclusion bodies.Results: Here we report that the intestine of the zebrafish goosepimples (gsp)/myosin Vb (myoVb) mutant show severe reduction in the intestinal folds-structures similar to mammalian villi. The loss of folds is further correlated with changes in the shape of enterocytes. In a striking similarity with MVID patients, zebrafish gsp/myoVb mutant larvae exhibit microvillus atrophy, microvillus inclusions and accumulation of secretory material in enterocytes.Conclusion: We propose that the zebrafish gsp/myoVb mutant is a valuable model to study the pathophysiology of MVID. Owing to the advantages of zebrafish in screening libraries of small molecules, the gsp mutant will be an appropriate tool to identify compounds that would alleviate the formation of microvillus inclusions and have therapeutic value.

Voltage-Sensing Phosphatase (VSP) Regulates Endocytosis-Dependent Nutrient Absorption in Chordate Enterocytes

“…This age also corresponds to the period when higher mortality was observed in Dr-VSP -/larvae (Figure 5A). We measured key parameters that represent cell morphology: central height, apical width, basal width, and microvillus length (Sidhaye et al, 2016). The measurement revealed that Dr-VSP -/-LREs were significantly shorter in all other parameters except for the microvillus length (Figure 7B).…”

“…Quantification of cellular dimensions of LREs was done based on the previous procedure (Sidhaye et al, 2016). We performed immunostaining with Acti-stain TM 488 Phalloidin (Cytoskeleton, USA) and anti-Na + -K + ATPase Antibody (Ab76020; Abcam, USA) to outline individual cell (Figure 7A).…”