Expression of carbonic anhydrase IV in mouse placenta

2001 ◽  
Vol 280 (2) ◽  
pp. R365-R375 ◽  
Author(s):  
Orna Rosen ◽  
Carlos Suarez ◽  
Victor L. Schuster ◽  
Luc P. Brion
Keyword(s):  
Carbonic Anhydrase ◽  
Rabbit Kidney ◽  
Developmentally Regulated ◽  
Western Blots ◽  
Immunoreactive Protein ◽  
Mouse Placenta ◽  
Weak Staining ◽  
Microsomal Membranes ◽  
Membrane Bound ◽  
Carbonic Anhydrase Iv

Carbonic anhydrase (CA) facilitates acid-base transport in several tissues. Acidosis upregulates membrane-bound SDS-resistant hydratase activity in various tissues and CA IV mRNA in rabbit kidney. This study was designed to assess whether the expression of membrane-bound CA IV isozyme in mouse placenta is regulated developmentally and by maternal ammonium chloride loading at the end of pregnancy. For this purpose we used Northern blot analysis, Western blots of microsomal membranes, and immunocytochemistry. The expression of CA IV mRNA on Northern blots tripled from day 11 to day 15 and then remained stable until the end of pregnancy. Expression of CA IV immunoreactive protein on Western blot tripled from day 11 to day 15 and decreased almost to baseline by day 19. Strong staining for CA IV was detected by immunocytochemistry in labyrinthine trophoblast, in the endodermal layer of the yolk sac (both intra- and extraplacental) and in the uterine epithelium. Weak staining was observed in most fetal endothelial cells at 11 days but not later in gestation. Maternal acidosis did not upregulate the expression of CA IV mRNA or CA IV immunoreactive protein. Thus CA IV expression in mouse placenta is developmentally regulated. Maternal acidosis during the last quarter of pregnancy does not upregulate CA IV mRNA or CA IV immunoreactive protein.

Expression of membrane-associated carbonic anhydrase isoforms IV, IX, XII, and XIV in the rabbit: induction of CA IV and IX during maturation

2005 ◽  
Vol 288 (5) ◽  
pp. R1256-R1263 ◽  
Author(s):  
Jeffrey M. Purkerson ◽  
George J. Schwartz
Keyword(s):  
Skeletal Muscle ◽  
Carbonic Anhydrase ◽  
Gall Bladder ◽  
Plasma Membranes ◽  
Rabbit Kidney ◽  
Α Subunit ◽  
Anion Transporters ◽  
Ca Ix ◽  
Membrane Bound ◽  
Physiological Demands

Several carbonic anhydrase (CA) isoforms are associated with plasma membranes. It is probable that these enzymes interact with anion transporters to facilitate the movement of HCO3− into or out of the cell. A better knowledge of CA isoform expression in a given tissue would facilitate a systematic examination of any associations with such transporters. We examined the expression of CAs IV, IX, XII, and XIV mRNAs in rabbit tissues, including kidney, heart, lung, skeletal muscle, liver, pancreas, gall bladder, stomach, small intestine, colon, and spleen, using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). CA IV mRNA was mainly in kidney, heart, lung, colon, and gall bladder. CA IX mRNA was restricted to stomach, gall bladder, duodenum, and early jejunum. CA XII mRNA was found in kidney and colon. CA XIV mRNA was localized to heart, lung, skeletal muscle, and liver. The data indicate that there are different patterns of CA expression in various tissues: CA IX was expressed in the proximal gastrointestinal tract, whereas CA XII and CA IV were more distal. CA IV and CA XII are important kidney isoforms. CA XIV was abundant in metabolically active tissues such as liver, heart, lung, and skeletal muscle. Some significant species differences were noted in the expression of some of these isoforms; for example, CA XIV is not expressed in rabbit kidney, despite being abundant in mouse kidney. Maturational studies showed that the expression of CA IX mRNA and protein increased markedly with weaning (∼3–4 postnatal wk) and was well correlated with the maturational expression of the α-subunit of the gastric H+,K+-ATPase, suggesting that function of CA IX and the gastric H+ pump might be linked in the digestion of adult foodstuffs. The unique pattern of membrane-bound CA isoforms suggests different functional associations with transporters, depending on the physiological demands on the tissue.

Membrane-bound carbonic anhydrase in the heart

1992 ◽  
Vol 262 (2) ◽  
pp. H577-H584 ◽  
Author(s):  
W. Bruns ◽  
G. Gros
Keyword(s):  
Carbonic Anhydrase ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Integral Membrane Protein ◽  
Rabbit Heart ◽  
Microsomal Membranes ◽  
Membrane Bound ◽  
The One ◽  
Triton X

Microsomal membranes from bovine heart homogenates were subfractionated by density gradient centrifugation. Fractions with high levels of a sarcolemmal (SL) marker are enriched in specific carbonic anhydrase (CA) activity up to ninefold compared with the microsomes. Fractions with high levels of a sarcoplasmic reticulum marker and a mitochondrial marker, respectively, exhibit specific CA activities that are similar to the one found in the microsomes. Determination of cytosolic markers reveals that the CA activity in the SL fraction is not due to contamination by cytosolic CA, and it is shown by Triton X-114 phase separation that the CA activity is due to an integral membrane protein. In cryosections from rabbit heart the SL region of cardiomyocytes is stained by the fluorescent CA inhibitor dansylsulfonamide. Intracellular staining occurs also, with a pattern suggesting the presence of CA associated with intracellular membranes. Although it cannot be excluded that there is a contribution by endothelial membranes, it appears likely that most CA of the heart is bound to the SL. The possible involvement of the enzyme in extracellular proton buffering is discussed.

scholarly journals Expression of Carbonic Anhydrase IV in Mouse Placenta

1999 ◽  
Vol 45 (4, Part 2 of 2) ◽  
pp. 60A-60A
Author(s):  
Orna Rosen ◽  
Carlos Suarez ◽  
Luc P Brion
Keyword(s):  
Carbonic Anhydrase ◽  
Mouse Placenta ◽  
Carbonic Anhydrase Iv

scholarly journals Immunohistochemical localization of carbonic anhydrase IV in capillaries of rat and human skeletal muscle.

1994 ◽  
Vol 42 (9) ◽  
pp. 1229-1236 ◽  
Author(s):  
S Sender ◽  
G Gros ◽  
A Waheed ◽  
G S Hageman ◽  
W S Sly
Keyword(s):  
Carbonic Anhydrase ◽  
Immunohistochemical Localization ◽  
Specific Staining ◽  
Atpase Reaction ◽  
Rabbit Igg ◽  
Sds Page ◽  
Membrane Bound ◽  
Carbonic Anhydrase Iv ◽  
Von Willebrand ◽  
Willebrand Factor

We used polyclonal antisera raised in rabbits against membrane-bound rat lung and human lung carbonic anhydrase (CA) IV in immunofluorescence studies to stain cryosections of rat soleus and extensor digitorum longus (EDL) and several human skeletal muscles. There was strong specific staining of capillaries in all muscles investigated. Several techniques were applied to verify this result. (a) Serial sections were either incubated with anti-CA IV/FITC or processed for endothelial ATPase reaction. There was precise co-localization of antibody marked structures and ATPase stained capillaries. (b) Human muscle sections were double stained with anti-CA IV/TRITC and anti-von Willebrand factor (vWF)/FITC. vWF, a capillary marker, and CA IV were localized at identical sites. (c) The CAIV was released from capillaries by treatment with phosphatidylinositol specific phospholipase C, suggesting that the enzyme is anchored to the endothelial cell membrane via a phosphatidylinositolglycan anchor. (d) A rat hindlimb was perfused with diluted antiserum. Cryosections of perfused soleus and EDL processed for anti-rabbit IgG/FITC staining showed clear fluorescence associated with capillaries, indicating that the antigen was accessible from the capillary lumen. (e) Immune complexes formed during antiserum perfusion as described in d were precipitated from muscle homogenates. SDS-PAGE followed by immunoblotting showed that the predominant portion of total muscle CA IV was bound in these complexes and therefore must be located intravascularly.

Maturation of carbonic anhydrase IV expression in rabbit kidney

2001 ◽  
Vol 280 (5) ◽  
pp. F895-F903 ◽  
Author(s):  
Cornelia A. Winkler ◽  
Ann M. Kittelberger ◽  
Richard H. Watkins ◽  
William M. Maniscalco ◽  
George J. Schwartz
Keyword(s):  
Carbonic Anhydrase ◽  
Proximal Tubule ◽  
Collecting Duct ◽  
High Rate ◽  
Rabbit Kidney ◽  
Mrna Levels ◽  
Intercalated Cells ◽  
Collecting Ducts ◽  
Medullary Collecting Duct ◽  
Carbonic Anhydrase Iv

Carbonic anhydrase (CA) IV facilitates renal acidification by catalyzing the dehydration of luminal H2CO3. CA IV is expressed in proximal tubules, medullary collecting ducts, and A-intercalated cells of the mature rabbit kidney (Schwartz GJ, Kittelberger AM, Barnhart DA, and Vijayakumar S. Am J Physiol 278: F894–F904, 2000). In view of the maturation of HCO[Formula: see text] transport in the proximal tubule and collecting duct, the ontogeny of CA IV expression was examined. During the first 2 wk, CA IV mRNA was expressed in maturing cortex and medulla at ∼20% of adult levels. The maturational increase was gradual in cortex over 3–5 wk of age but surged in the medulla, so that mRNA levels appeared higher than those in the adult medulla. In situ hybridization showed very little CA IV mRNA at 5 days, with increases in deep cortex and medullary collecting ducts by 21 days. Expression of CA IV protein in the cortex and medulla was minimal at 3 days of age but then apparent in the juxtamedullary region, A-intercalated cells and medullary collecting ducts by 18 days; there was little labeling of the proximal straight tubules of the medullary rays. Thus CA IV expression may be regulated to accommodate the maturational increase in HCO[Formula: see text] absorption in the proximal tubule. In the medullary collecting duct, there is a more robust maturation of CA IV mRNA and protein, commensurate with the high rate of HCO[Formula: see text] absorption in the neonatal segment.

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