Heparan sulfate proteoglycans (HSPGs) are constituents of the cell surface and extracellular matrix and are vital for various activities within the cell. The N-deacetylase/N-sulfotransferase (heparin glucosaminyl) family of enzymes, or NDST, modifies heparan sulfate (HS) by catalyzing both the N-deacetylation and the N-sulfation of N-acetylglucosamine residues. In zebrafish, a single ndst3 gene is an orthologue of both mammalian NDST3 and NDST4 genes. The role of ndst3 in zebrafish development has not been investigated and such study may provide insight into the role(s) of both mammalian orthologues. Here, we characterized expression of ndst3 during early development in zebrafish and found it to be predominately neuronal. We found that expression of ndst3 is sensitive to Wnt signaling manipulation, with stimulation of the Wnt pathway resulting in robust expansion of ndst3 expression domains. Finally, using CRISPR/Cas9 genome editing, we mutagenized the ndst3 gene and isolated an allele, ndst3^nu20, resulting in a frameshift and premature protein truncation. We discovered Ndst3 is not essential for zebrafish survival as ndst3^nu20 homozygous mutants are viable and fertile.

硫酸乙酰肝素蛋白聚糖 (HSPG) 是细胞表面和细胞外基质的成分，对于细胞内的各种活动至关重要。N-脱乙酰酶/N-磺基转移酶（肝素葡糖胺基）家族（NDST）通过催化 N-乙酰葡糖胺残基的 N-脱乙酰化和 N-硫酸化来修饰硫酸乙酰肝素 (HS)。在斑马鱼中，单个ndst3基因是哺乳动物NDST3和NDST4基因的直系同源物。ndst3在斑马鱼发育中的作用尚未得到研究，此类研究可能有助于深入了解两种哺乳动物直系同源物的作用。在这里，我们表征了斑马鱼早期发育过程中ndst3的表达，发现它主要是神经元的表达。我们发现ndst3的表达对 Wnt 信号传导操作敏感，刺激 Wnt 通路会导致ndst3表达域的强劲扩展。最后，使用 CRISPR/Cas9 基因组编辑，我们对ndst3基因进行诱变并分离出一个等位基因ndst3 ^nu20，导致移码和过早的蛋白质截短。我们发现 Ndst3 对于斑马鱼的生存并不是必需的，因为ndst3 ^nu20纯合突变体是可行且可繁殖的。