Abstract:
:The polarity of retinal pigmented epithelia (RPE) from chicken embryos was studied in primary cell culture. Since cultured RPE approximates the morphological polarity of RPE in vivo, we investigated whether this polarity extends to the distribution of plasma membrane proteins that are peculiar to RPE. In contrast to other epithelia, the Na+,K(+)-ATPase of RPE is located in the apical rather than basolateral plasma membrane. To examine this property, we cultured RPE on extracellular matrix-coated filters. Primary cultures were compared to embryonic RPE in situ using electron microscopy and indirect immunofluorescence of frozen sections. The viability and morphology of RPE was improved by using a serum-free medium containing a bovine pituitary extract in conjunction with an extracellular matrix coating derived from Engelbreth-Holm-Swarm tumors. Cultured RPE mimicked the morphology of RPE in vivo with microvilli and junctional complexes on the apical pole and infoldings along the basolateral plasma membrane. Functional tight junctions formed as demonstrated by an EDTA-sensitive, transepithelial electrical resistance, and by the retention of [3H]inulin added to the apical chamber. In 2 hr, only 4-6% of the [3H]inulin crossed the monolayer, compared to 24% in control filters. Despite these features of polarity, the Na+,K(+)-ATPase was detected in both apical and basolateral membranes by immunofluorescence. In embryonic eyes in which the neural retina was removed, the Na+,K(+)-ATPase was confined to the apical membrane. In addition, the polarity of cultured RPE was probed with vesicular stomatitis virus. In contrast to other epithelia, budding virus particles were observed emerging from the apical, as well as basolateral, domain further suggesting the cultured cells were only partially polarized. These data indicate that structural criteria are inadequate to determine if cultured RPE have become polarized in the same manner as the epithelium in vivo.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
Rizzolo LJdoi
10.1016/0014-4835(90)90156-osubject
Has Abstractpub_date
1990-10-01 00:00:00pages
435-46issue
4eissn
0014-4835issn
1096-0007pii
0014-4835(90)90156-Ojournal_volume
51pub_type
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journal_title:Experimental eye research
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journal_title:Experimental eye research
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pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:Experimental eye research
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更新日期:2018-12-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(83)90156-2
更新日期:1983-09-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1993.1186
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journal_title:Experimental eye research
pub_type: 杂志文章
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1996.0147
更新日期:1996-11-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/0014-4835(91)90064-l
更新日期:1991-05-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章,评审
doi:10.1016/j.exer.2011.10.014
更新日期:2012-02-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2009.01.011
更新日期:2009-06-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1006/exer.1997.0315
更新日期:1997-07-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
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更新日期:1983-02-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/s0014-4835(87)80144-6
更新日期:1987-08-01 00:00:00
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journal_title:Experimental eye research
pub_type: 杂志文章
doi:10.1016/j.exer.2007.04.005
更新日期:2007-08-01 00:00:00