Sannes P L, Burch K K, Khosla J, McCarthy K J, Couchman J R
Department of Anatomy, North Carolina State University, College of Veterinary Medicine, Raleigh 27606.
Am J Respir Cell Mol Biol. 1993 Mar;8(3):245-51. doi: 10.1165/ajrcmb/8.3.245.
Histologic preparations of lungs from 1-, 5-, 10-, 18-, and 25-day-old postnatal and adult rats were examined immunohistochemically with antibodies specific against chondroitin sulfate (CS), basement membrane chondroitin sulfate proteoglycan (BM-CSPG), heparan sulfate proteoglycan (HSPG), entactin, and laminin. A monoclonal antibody specific for the glycosaminoglycan portion (CS) of CSPG and a monoclonal antibody against the core protein of CSPG were used in an immunoperoxidase sequence to stain extracellular matrix (ECM) components of pulmonary basement membranes (BMs). Anti-CS stained airway BM strongly and alveolar BM weakly in the adult rat lung, as well as in vascular and airway adventitia. In developing lungs, immunoreactivity was strong in all ECM sites, including BM, at day 1 postnatal, and progressively diminished thereafter except in vascular and airway adventitia. Anti-CSPG stained alveolar, airway, and vascular BMs, in addition to smooth muscle external laminae (EL), in the adult and developing rat. Immunostaining for CSPG required hyaluronidase digestion, whereas CS staining was lost with the same treatment. A polyclonal antibody to the core protein of HSPG was found to be similarly distributed to CSPG by immunoperoxidase staining in adult and developing rat lungs, with the notable exception that little immunoreactivity for HSPG was found in smooth muscle EL. Commercially obtained polyclonal antibodies to entactin and laminin gave immunostaining comparable to that seen with CSPG, except that entactin showed particular affinity for EL. These results offer a more detailed perspective on previous survey observations of CSPG, HSPG, and entactin in the rat lung, and describe the immunoreactivity of CS for the first time.(ABSTRACT TRUNCATED AT 250 WORDS)
使用针对硫酸软骨素(CS)、基底膜硫酸软骨素蛋白聚糖(BM-CSPG)、硫酸乙酰肝素蛋白聚糖(HSPG)、巢蛋白和层粘连蛋白的特异性抗体,对出生后1天、5天、10天、18天、25天的幼鼠以及成年大鼠的肺组织学标本进行免疫组织化学检查。使用一种针对CSPG糖胺聚糖部分(CS)的单克隆抗体和一种针对CSPG核心蛋白的单克隆抗体,通过免疫过氧化物酶序列来染色肺基底膜(BM)的细胞外基质(ECM)成分。抗CS抗体在成年大鼠肺以及血管和气道外膜中强烈染色气道BM,而对肺泡BM染色较弱。在发育中的肺中,出生后第1天所有ECM部位(包括BM)的免疫反应性都很强,此后除血管和气道外膜外逐渐减弱。抗CSPG抗体在成年和发育中的大鼠中除了平滑肌外板(EL)外,还对肺泡、气道和血管BM进行染色。CSPG的免疫染色需要用透明质酸酶消化,而相同处理会使CS染色消失。通过免疫过氧化物酶染色发现,一种针对HSPG核心蛋白的多克隆抗体在成年和发育中的大鼠肺中的分布与CSPG相似,但值得注意的是,在平滑肌EL中几乎没有发现HSPG的免疫反应性。商业获得的针对巢蛋白和层粘连蛋白的多克隆抗体产生的免疫染色与CSPG相当,只是巢蛋白对EL表现出特别的亲和力。这些结果为先前对大鼠肺中CSPG、HSPG和巢蛋白的调查观察提供了更详细的视角,并首次描述了CS的免疫反应性。(摘要截短至250字)