Expression of Na(+)/K(+)-ATPase alpha subunit isoforms in rat salivary glands: occurrence of sense and antisense RNAs of the alpha3 isoform in the sublingual gland.
We examined the expression of Na(+)/K(+)-ATPase alpha-subunit isoforms in rat salivary glands by RT-PCR. Isoform alpha1 was expressed strongly in all three major salivary glands. The alpha2 isoform was expressed in both submandibular gland (SMG) and sublingual gland (SLG) but faintly in the parotid gland (PG). The alpha3 was detected only in the SLG, and the alpha3 mRNA in the SLG was 1/8 of its level in the brain. Na(+)/K(+)-ATPase alpha3 isoform in the SLG, was localized predominantly on the basolateral plasma membranes in serous cells by immunohistochemical method. We also found the presence of natural antisense RNA of the alpha3 isoform in rat SLG: the 1st-strand cDNA prepared with gene-specific forward primers targeted to the CDS region and to the promoter region of the alpha3 gene in place of oligo(dT) or gene-specific reverse primers produced reasonable PCR products corresponding to the alpha3 cDNA sequence by the subsequent PCR reaction. Synthesis of the 1st-strand cDNA with the gene-specific forward primers was prevented by RNase digestion of the total RNA preparation, indicating that the PCR products in the RT-PCR system were not due to the contaminated genomic DNA, if any. The alpha3 protein level in rat SLG increased with aging, and levels of both alpha3 mRNA (sense RNA) and alpha3 antisense RNA were higher in SLGs of aged rats than in those of young rats, respectively.
Kurihara
Kinji
K
Division of Physiology, Meikai University, School of Dentistry, Sakado, Saitama, Japan. kkinji@dent.meikai.ac.jp
Nakanishi
Nobuo
N
Amano
Osamu
O
Tonosaki
Keiichi
K
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression / Q000502:physiology; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008297:Male; D010306:Parotid Gland / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D016372:RNA, Antisense / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D053651:Receptors, Interleukin-9 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012469:Salivary Glands / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D013361:Sublingual Gland / Q000378:metabolism*
D017931:DNA Primers; D007527:Isoenzymes; D016372:RNA, Antisense; D053651:Receptors, Interleukin-9; D012313:RNA; D000254:Sodium-Potassium-Exchanging ATPase