The plasma membrane carbonic anhydrase in murine hepatocytes identified as isozyme XIV.
BACKGROUND
Biochemical and histochemical studies have both previously indicated plasma membrane-associated carbonic anhydrase (CA) activity in hepatocytes which has been assumed to be CA IV. However, immunohistochemical data did not support this assignment. Recent northern blotting results indicated the presence of mRNA for the most recently discovered membrane-bound CA isozyme, CA XIV, in the liver. The present study was designed to examine whether CA XIV could contribute to the CA activity described in the hepatocytes.


METHODS
Tissue samples from mouse liver were subjected to immunohistochemical staining using the antibodies raised against recombinant mouse CA XIV and CA IV. RT-PCR and western blotting were also performed for CA XIV.


RESULTS
A strong immunofluorescent signal was observed in the plasma membrane of mouse hepatocytes. Although CA XIV was expressed on both the apical and basolateral surfaces, the staining was more prominent at the apical (canalicular) membrane domain. The expression of CA XIV in the liver was confirmed by RT-PCR and western blotting.


CONCLUSIONS
The presence of CA XIV in the hepatocyte plasma membrane places this novel enzyme at a strategic site to control pH regulation and ion transport between the hepatocytes, sinusoids and bile canaliculi.
Parkkila
Seppo
S
Department of Clinical Chemistry, 90014 University of Oulu, Finland. seppo.parkkila@uta.fi
Kivelä
Antti J
AJ
Kaunisto
Kari
K
Parkkila
Anna-Kaisa
AK
Hakkola
Jukka
J
Rajaniemi
Hannu
H
Waheed
Abdul
A
Sly
William S
WS
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D016466:CHO Cells; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D003106:Colon / Q000737:chemistry / Q000201:enzymology; D006224:Cricetinae; D022781:Hepatocytes / Q000201:enzymology*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D007583:Jejunum / Q000737:chemistry / Q000201:enzymology; D008099:Liver / Q000737:chemistry / Q000201:enzymology; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012313:RNA / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000906:Antibodies; D007527:Isoenzymes; D008565:Membrane Proteins; D012313:RNA; D002256:Carbonic Anhydrases; C466621:carbonic anhydrase XIV