Sugahara Daisuke, Kawakami Hayato, Akimoto Yoshihiro
Laboratory of Microscopic Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
PLoS One. 2025 Jul 8;20(7):e0326157. doi: 10.1371/journal.pone.0326157. eCollection 2025.
The colonic mucin layer, comprising highly glycosylated mucin proteins, is crucial for maintaining colonic health. Its region-specific glycosylation patterns are indispensable for adapting to distinct physiological and microbial environments along the colon, thus ensuring appropriate mucin layer function. However, the mechanisms underlying this region-specific glycosylation remain unknown. Here, using fluorescence-based immunohistological analyses of the colon from experimental mice, we demonstrated that along with contribution of goblet cells, as conventionally believed, mucin glycosylation involves deep crypt secretory (DCS) cells, a specialized mucin-producing cell population in the colon. Based on cKit/CD117 as a DCS cell marker, DCS and goblet cells are inversely distributed along the mouse colon: DCS cells predominate proximally, constituting nearly 70% of mucin-producing cells, whereas goblet cells are more abundant distally, indicating a dynamic shift in the predominant mucin-producing cell population along the colon. Immunofluorescence staining revealed that DCS cells produce distinctive mucin-glycans, including those with the Core3-glycan motif that exhibit region-specific distributions in the mucin layer. We found that the gradient distribution of DCS cells predominantly shapes their region-specific distribution, whereas the inverse distribution of goblet cells corresponds to the distal distribution of sulfated and sialylated glycans. Furthermore, the in situ Proximity Ligation Assay for specifically detecting Muc2 with distinct glycosylation, revealed that DCS and goblet cells produce different types of α1,2-fucosylated glycans on Muc2, indicating that the shift in the predominant mucin-producing cells drives region-specific α1,2-fucosylation on Muc2 across colonic regions. Although DCS cells are implicated in supporting the stem cell niche, their involvement in mucin production was unclear. We highlight the critical role of DCS cells in establishing regional glycosylation patterns. Our findings provide new insights into the cellular basis of mucin glycosylation, as well as their potential impact on colonic health and disease susceptibility in specific colonic regions.
结肠黏液层由高度糖基化的黏蛋白组成，对维持结肠健康至关重要。其区域特异性糖基化模式对于适应结肠不同的生理和微生物环境不可或缺，从而确保黏液层功能正常。然而，这种区域特异性糖基化的潜在机制仍不清楚。在此，我们通过对实验小鼠结肠进行基于荧光的免疫组织学分析，证明了与传统认知中杯状细胞的作用一样，黏蛋白糖基化涉及深部隐窝分泌（DCS）细胞，这是结肠中一种专门的黏蛋白产生细胞群体。基于cKit/CD117作为DCS细胞标志物，DCS细胞和杯状细胞沿小鼠结肠呈反向分布：DCS细胞在近端占主导，占黏蛋白产生细胞的近70%，而杯状细胞在远端更为丰富，这表明沿结肠主要黏蛋白产生细胞群体发生了动态变化。免疫荧光染色显示，DCS细胞产生独特的黏蛋白聚糖，包括具有Core3-聚糖基序的那些，它们在黏液层中呈现区域特异性分布。我们发现，DCS细胞的梯度分布主要塑造了它们的区域特异性分布，而杯状细胞的反向分布与硫酸化和唾液酸化聚糖的远端分布相对应。此外，用于特异性检测具有不同糖基化的Muc2的原位邻近连接分析表明，DCS细胞和杯状细胞在Muc2上产生不同类型的α1,2-岩藻糖基化聚糖，这表明主要黏蛋白产生细胞的转变驱动了跨结肠区域Muc2上的区域特异性α1,2-岩藻糖基化。尽管DCS细胞被认为与支持干细胞生态位有关，但其在黏蛋白产生中的作用尚不清楚。我们强调了DCS细胞在建立区域糖基化模式中的关键作用。我们的研究结果为黏蛋白糖基化的细胞基础及其对特定结肠区域结肠健康和疾病易感性的潜在影响提供了新的见解。
