Amyloid precursor protein-b facilitates cell adhesion during early development in zebrafish.

Scientific reports (2020-06-25)
Rakesh Kumar Banote, Jasmine Chebli, Tuğçe Munise Şatır, Gaurav K Varshney, Rafael Camacho, Johan Ledin, Shawn M Burgess, Alexandra Abramsson, Henrik Zetterberg

Understanding the biological function of amyloid beta (Aβ) precursor protein (APP) beyond its role in Alzheimer's disease is emerging. Yet, its function during embryonic development is poorly understood. The zebrafish APP orthologue, Appb, is strongly expressed during early development but thus far has only been studied via morpholino-mediated knockdown. Zebrafish enables analysis of cellular processes in an ontogenic context, which is limited in many other vertebrates. We characterized zebrafish carrying a homozygous mutation that introduces a premature stop in exon 2 of the appb gene. We report that appb mutants are significantly smaller until 2 dpf and display perturbed enveloping layer (EVL) integrity and cell protrusions at the blastula stage. Moreover, appb mutants surviving beyond 48 hpf exhibited no behavioral defects at 6 dpf and developed into healthy and fertile adults. The expression of the app family member, appa, was also found to be altered in appb mutants. Taken together, we show that appb is involved in the initial development of zebrafish by supporting the integrity of the EVL, likely by mediating cell adhesion properties. The loss of Appb might then be compensated for by other app family members to maintain normal development.

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Penicillin-Streptomycin, Solution stabilized, with 10,000 units penicillin and 10 mg streptomycin/mL, 0.1 μm filtered, BioReagent, suitable for cell culture
L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
TRI Reagent®, For processing tissues, cells cultured in monolayer or cell pellets
Ionomycin calcium salt from Streptomyces conglobatus, powder, ≥98% (HPLC)
Anti-α-Tubulin antibody, Mouse monoclonal, clone DM1A, purified from hybridoma cell culture
Anti-β-Catenin antibody produced in rabbit, whole antiserum