Instituto de Histologia e Embriologia, Faculdade de Medicina, Universidade de Coimbra; Hospitais da Universidade de Coimbra .
Eur J Histochem. 2019 Sep 11;63(3):3040. doi: 10.4081/ejh.2019.3040.
In mammals, the alveolarization process develops predominantly after birth. Airway cells display a complex assemblage of glycans on their surface. These glycans, particularly terminal glycan extensions, are important effective carriers of information that change during the differentiation process. Nevertheless, few systematic data are reported about the cell surface sugar residue content during post-natal lung development. In the present work, we aimed to identify and semi-quantify N-acetylgalactosamine (GalNAc)/galactose (Gal) residues on the bronchioloalveolar cell surface in rat lung sections from 1-, 4-, 8- day old and adult animals and link these data with the lung glycocalyx composition. Horseradish peroxidase-conjugated lectin from Glycine max (soybean agglutinin, SBA) was used, and light microscopy methodologies were performed. SBA labelling intensity was studied before and after sialidase pre-treatment, at one-, four- and eight-day-old animals and adult animals. For semi-quantitative evaluation of SBA binding intensity, two investigators performed the analysis independently, blinded to the type of experiment. Reactivity of the lectin was assessed in bronchiolar and respiratory portion/alveolar epithelial cell surfaces. We evidenced a stronger positive reaction when lung sections were pre-treated with neuraminidase before incubation with the lectin in one- and four-day-old animals and adult animals. These results were not so manifest in eight-day-old animals. This binding pattern, generally points towards the presence of terminal but mainly sub-terminal GalNAc/Gal residues probably capped by sialic acids on the rat bronchiolar/respiratory tract epithelial cells. As this glycan extension is common in O- and N-glycans, our results suggest that these glycan classes can be present in bronchioloalveolar cells immediately after birth and exist during the postnatal period. The results observed in eight-day-old rat lung sections may be due to the dramatic lung morphologic changes and the possible underlying biological mechanisms that occur during this age-moment.
在哺乳动物中,肺泡发育过程主要在出生后进行。气道细胞在其表面显示出复杂的聚糖组合。这些糖,特别是末端糖延伸,是在分化过程中发生变化的信息的重要有效载体。然而,关于出生后肺发育过程中细胞表面糖残基含量的系统数据很少。在本工作中,我们旨在鉴定和半定量大鼠肺组织中 1、4、8 天龄和成年动物支气管肺泡细胞表面的 N-乙酰半乳糖胺(GalNAc)/半乳糖(Gal)残基,并将这些数据与肺糖萼组成联系起来。使用了辣根过氧化物酶偶联的 Glycine max(大豆凝集素,SBA),并进行了光学显微镜方法学研究。在 1、4 和 8 天龄和成年动物中,在唾液酸酶预处理前后研究了 SBA 标记强度。为了对半定量评估 SBA 结合强度,两位研究人员独立进行了分析,对实验类型不了解。评估了凝集素在细支气管和呼吸部分/肺泡上皮细胞表面的反应性。在 1 天龄和 4 天龄以及成年动物中,在与凝集素孵育之前,用神经氨酸酶预处理肺切片时,我们证明了更强的阳性反应。在 8 天龄动物中,这些结果并不明显。这种结合模式通常表明存在末端但主要是次末端的 GalNAc/Gal 残基,可能在大鼠细支气管/呼吸道上皮细胞上被唾液酸糖帽。由于这种聚糖延伸在 O-和 N-聚糖中很常见,我们的结果表明这些糖类可以在出生后立即存在于支气管肺泡细胞中,并在出生后期间存在。在 8 天龄大鼠肺切片中观察到的结果可能是由于在这个年龄段肺形态发生剧烈变化和可能发生的潜在生物学机制。