Chambard M, Verrier B, Gabrion J, Mauchamp J
J Cell Biol. 1983 Apr;96(4):1172-7. doi: 10.1083/jcb.96.4.1172.
When cultured in collagen gel-coated dishes, thyroid cells organized into polarized monolayers. The basal poles of the cells were in contact with the collagen gel, whereas the apical surfaces were facing the culture medium. Under these culture conditions, thyroid cells do not concentrate iodide nor respond to acute stimulation by thyroid-stimulating hormone (TSH). To allow the free access of medium components to the basal poles, the gel was detached from the plastic dish and allowed to float in the culture medium. After release of the gel, the iodide concentration and acute response to TSH stimulation were restored. Increased cAMP levels, iodide efflux, and formation of apical pseudopods were observed. When the thyroid cells are cultured on collagen-coated Millipore filters glued to glass rings, the cell layer separates the medium in contact with the apical domain of the plasma membrane (inside the ring) from that bathing the basolateral domain (outside the ring). Iodide present in the basal medium was concentrated in the cells, whereas no transport was observed when iodide was added to the luminal side. Similarly, an acute effect of TSH was observed only when the hormone was added to the basal medium. These results show that the iodide concentration mechanism and the TSH receptor-adenylate cyclase complex are present only on the basolateral domain of thyroid cell plasma membranes.
当在涂有胶原凝胶的培养皿中培养时,甲状腺细胞会组织形成极化的单层。细胞的基底部与胶原凝胶接触,而顶端表面则朝向培养基。在这些培养条件下,甲状腺细胞不会摄取碘化物,也不会对促甲状腺激素(TSH)的急性刺激产生反应。为了使培养基成分能够自由接触细胞基底部,将凝胶从塑料培养皿上分离,使其漂浮在培养基中。凝胶分离后,碘化物摄取能力和对TSH刺激的急性反应得以恢复。观察到细胞内cAMP水平升高、碘外流以及顶端伪足形成。当甲状腺细胞在粘贴于玻璃环上的胶原包被的密理博滤膜上培养时,细胞层将与质膜顶端结构域接触的培养基(环内)与浸润基底外侧结构域的培养基(环外)分隔开来。基底培养基中的碘化物会在细胞内聚集,而当碘化物添加到管腔侧时则未观察到转运现象。同样,只有当激素添加到基底培养基中时才观察到TSH的急性效应。这些结果表明,碘化物摄取机制和TSH受体 - 腺苷酸环化酶复合物仅存在于甲状腺细胞质膜的基底外侧结构域。