Carbonic anhydrase gene expression in CA II-deficient (Car2-/-) and CA IX-deficient (Car9-/-) mice.
Using real-time PCR and immunohistochemistry, we have examined the expression of carbonic anhydrase isozymes (CA) I, II, III, IV, IX, XII, XIII and XIV in the brain, kidney, stomach and colon of the wild-type, CA II-deficient (Car2-/-), and CA IX deficient (Car9-/-) mice. The expression of Car4, Car12, Car13 and Car14 mRNAs did not show any significant deviations between the three groups of mice, whereas both groups of CA deficient mice showed decreased expression levels of Car1 in the colon and Car3 in the kidney. The Car2 mRNA level was greatly reduced but not completely abolished in all four tissues from the Car2-/- mice in which no CA II protein was expressed. Sequencing the Car2 cDNA isolated from C57BL6 Car2-/- mice revealed two nucleotide differences from the wild-type C57BL6 mice. One is a silent polymorphism found in Car2 mRNA from wild-type DBA mice, which is the strain that provided the original mutagenized chromosome. The second change is a mutation that causes prematurely terminated translation at codon 155 (Gln155X). Car9 mRNA and CA IX protein expression levels were up-regulated about 2.5- and 3.6-fold, respectively, in the stomach of the Car2-/- mice. These results suggest that the loss of function of cytosolic CA II in the stomach of Car2-/- mice leads to up-regulation of an extracellular CA, namely CA IX, which is expressed on the cell surface of the gastric epithelium.
Pan
Peiwen
P
Institute of Medical Technology, University of Tampere and Tampere University Hospital, Biokatu 6, 33520 Tampere, Finland. peiwen.pan@uta.fi
Leppilampi
Mari
M
Pastorekova
Silvia
S
Pastorek
Jaromir
J
Waheed
Abdul
A
Sly
William S
WS
Parkkila
Seppo
S
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism; D024402:Carbonic Anhydrase II / Q000235:genetics* / Q000378:metabolism*; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D003106:Colon / Q000378:metabolism; D005260:Female; D005753:Gastric Mucosa / Q000378:metabolism; D018628:Gene Dosage; D007150:Immunohistochemistry; D007668:Kidney / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D017385:Sequence Homology; D015854:Up-Regulation
D012333:RNA, Messenger; D024402:Carbonic Anhydrase II; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases; C492796:Car9 protein, mouse