The Na ؉ ͞I ؊ symporter (NIS) is the plasma membrane protein that catalyzes active I ؊ transport in the thyroid, the first step in thyroid hormone biogenesis. The cDNA encoding NIS was recently cloned in our laboratory and a secondary structure model proposed, suggesting that NIS is an intrinsic membrane protein (618 amino acids; Ϸ65.2 kDa predicted molecular mass) with 12 putative transmembrane domains. Here we report the generation of a site-directed polyclonal anti-COOH terminus NIS antibody (Ab) that immunoreacts with a Ϸ87 kDa-polypeptide present in membrane fractions from a rat thyroid cell line (FRTL-5). The modelpredicted cytosolic-side location of the COOH terminus was confirmed by indirect immunof luorescence experiments using anti-COOH terminus NIS Ab in permeabilized FRTL-5 cells. Immunoreactivity was competitively blocked by the presence of excess synthetic peptide. Treatment of membrane fractions from FRTL-5 cells, Xenopus laevis oocytes, and COS cells expressing NIS with peptidyl N-glycanase F converted the Ϸ87 kDa-polypeptide into a Ϸ50 kDa-species, the same relative molecular weight exhibited by NIS expressed in E. coli. Anti-NIS Ab immunoprecipitated both the NIS precursor molecule (Ϸ56 kDa) and the mature Ϸ87 kDa form. Furthermore, a direct correlation between circulating levels of thyroidstimulating hormone and NIS expression in vivo was demonstrated.
In our experience polypropylene mesh used as a broad based tension-free sling was successful for treating all types of SUI. In our opinion technique and case selection have a bearing on outcomes.
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