A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3- reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Carbonic anhydrase II in the kidneys of normal and potassium-depleted rats using Western blot analysis and immuno-histochemistry. Western blot analysis demonstrated that CA II protein, ~30 kDa at molecular mass, was abundantly expressed in normal group. All potassium-depleted groups showed slightly increased CA II protein compared to normal group. In control group, immunoreactivity of CA II protein was detected in the entire collecting duct. Signal intensity was prominent in the intercalated cells and weak in the principal cells of the cortical collecting ducts. In potassium-depleted groups, the pattern of cellular labeling of CA II protein was identical to that of normal group, but the signal intensity was decreased in cortical collecting duct, markedly increased in the inner stripe of outer medullary and inner medullary collecting ducts, and unchanged in the outer stripe of outer medullary collecting duct. These results suggest that chronic hypokalemia impact the expression pattern of CA II protein depending the portion of the collecting duct.
Animals ; Blotting, Western ; Carbon ; Carbonic Anhydrase II ; Carbonic Anhydrases ; Hypokalemia ; Immunohistochemistry ; Kidney ; Rats

The carbonic anhydrase II (CA-II) is specifically expressed in oligodendrocytes, the cells responsible for myelination in the central nervous system. However no direct evidence on relationship between myelin formation and CA-II immunoreactivity has been described. The aims of these studies are to investigate the relationship between CA-II and myelination during cerebellar development of mouse. Myelin staining was found on postnatal (P) 14, and its intensity increased in proportion to developmental age. CA-II positive oligodendrocytes were observed in the white matter of cerebellum on P 14 day. CA-II positive oligoden-drocytes also occured in the granular layer and Purkinje cell layers in the later stage of dvelopment. The parallel development in the CA-II expression and myelination during development suggests that CA-II in oligoendrocyte play a role to myelination.
Animals ; Carbon* ; Carbonic Anhydrase II* ; Carbonic Anhydrases* ; Central Nervous System ; Cerebellum* ; Mice* ; Myelin Sheath ; Oligodendroglia

OBJECTIVETo investigate the expression of carbonic anhydrase II (CA2) in human testes and spermatozoa, and to compare the expressions of CA2 in ejaculated spermatozoa between normozoospermic and asthenozoospermic men.

METHODSThe localization of CA2 in human testes was observed by immunohistochemistry, and that in human sperm by immunofluorescence. Western blot was used to detect the expression of CA2 in the semen samples obtained from 16 normozoospermic and 16 asthenozoospermic volunteers.

RESULTSThe CA2 protein was shown to be localized in the tail of elongating spermatids by immunohistochemistry and in the flagellum of human sperm by immunofluorescence. Western blot revealed an obviously increased expression of CA2 in the spermatozoa of asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.84 +/- 0.32 vs 1.41 +/- 0.26, P < 0.05).

CONCLUSIONThe CA2 protein is expressed in the spermatogenic stage of elongating spermatids in human testes and localized in the sperm tail. The expression of CA2 is significantly increased in the spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.

Asthenozoospermia ; metabolism ; Carbonic Anhydrase II ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism ; Testis ; metabolism

The Mainz classification for renal cell tumors was introduced in 1986 and it's utility has been reported in several histogenetic and genetic studies of renal cell tumors. We present a study of 127 cases of renal cell tumors with clinicopathologic correlation, DNA content analysis, and histogenesis studied by histochemical and immunohistochemical staining. The 127 renal cell tumors classified by the Mainz classification were 87 clear cell, 17 chromophilic, 13 chromophobe and 3 sarcomatoid renal cell carcinomas, 5 oncocytomas and 2 adenomas. These subtypes showed significant correlation not with age, sex, Robson's stage, DNA ploidy or tumor recurrence but with nuclear grade (p=0.001) and tumor size (p=0.001). Hall's colloidal iron (p=0.002) and carbonic anhydrase II (p=0.013) stains, representing the origin of distal nephron especially of collecting duct, were significantly correlated with specific subtypes of renal cell tumors, especially chromophobe cell renal carcinoma. This study demonstrates that the Mainz classification suggests several morphologically different subtypes and variants of renal cell tumors and that some of them may have originated from the distal nephron, particularly from the collecting duct.
Adenoma ; Adenoma, Oxyphilic ; Carbonic Anhydrase II ; Carcinoma, Renal Cell ; Classification* ; Colloids ; Coloring Agents ; DNA* ; Iron ; Nephrons ; Ploidies ; Recurrence

OBJECTIVE: To investigate the changes of carbonic anhydrase (CA) activity and its mRNA expression in rats cochlea after noise exposure. METHOD: Twenty-four healthy Sprague Dawley rats were randomly allocated into 4 groups including 1 control group without contact of noise and 3 experimental groups exposed to a narrowband white noise of 4 kHz, 110 dB SPL 4h/d for 1 day, 1 week and 3 weeks, respectively. ABR thresholds were tested for each group at the time points described above. An immunohistochemical method was used to detect the expression of CA in the cochlea tissue. Differences in CAII mRNA expressions with and without noise exposure were examined using RT-PCR. RESULT: The thresholds of ABR in noise exposed cochlea were increased compared with the control cochlea (P < 0.01). CA activity and the CAII mRNA expression in noise exposed cochlea were decreased compared with the control cochlea (P < 0.01). The increase of ABR thresholds and decreases of CA activity and the CAII mRNA expression showed a time dependent pattern as the extension of duration exposed to noise. CONCLUSION Noise exposure could downregulate the CA activity and CAII mRNA expression in the cochlea. CA was proably involved in the pathogenesis of noise-induced hearing loss.
Animals ; Carbonic Anhydrase II ; genetics ; metabolism ; Cochlea ; enzymology ; Female ; Male ; Noise ; adverse effects ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

This study examined the anti-osteoclastogenic effects of baicalin on receptor activator of NF-kB ligand (RANKL)-induced RAW264.7 cells. Baicalin is a flavonoid that is produced by Scutellaria baicalensis and is known to have multiple biological properties, including antibacterial, anti-inflammatory and analgesic effects. The effects of baicalin on osteoclasts were examined by measuring 1) cell viability; 2) the formation of tartrate-resistant acid phosphatase (TRAP) (+) multinucleated cells; 3) RANK/RANKL signaling pathways and 4) mRNA levels of osteoclast-associated genes. Baicalin inhibited the formation of RANKL-stimulated TRAP (+) multinucleated cells and also suppressed the RANKL-stimulated activation of p-38, ERK, cSrc and AKT signaling. Baicalin also inhibited the RANKL-stimulated degradation of IkappaB in RAW264.7 cells. In addition, the RANKL-stimulated induction of NFATc1 transcription factors was found to be abrogated by this flavonoid. Baicalin was further found to decrease the mRNA expression of osteoclast-associated genes, including carbonic anhydrase II, TRAP and cathepsin K in the RAW264.7 cells. Our data thus demonstrate that baicalin inhibits osteoclastogenesis by inhibiting the RANKL-induced activation of signaling molecules and transcription factors in osteoclast precursors.
Acid Phosphatase ; Carbonic Anhydrase II ; Cathepsin K ; Flavonoids ; Isoenzymes ; NF-kappa B ; NFATC Transcription Factors ; Osteoclasts ; RNA, Messenger ; Scutellaria baicalensis ; Transcription Factors

Osteopetrosis is a rare genetic bone disease characterized by increased bone density but prone to breakage due to defective osteoclastic function. Among two primary types of autosomal dominant osteopetrosis (ADO), osteopetrosis type II is characterized by sclerosis of bones, predominantly involving the spine, the pelvis, and the skull base. Fragility of bones and dental abscess are leading complications. This report presents a case of osteopetrosis in a 52-years-old female, which was complicated by the development of cavernous sinus thrombophlebitis and meningitis. She was suffered from multiple fractures since one year ago. Laboratory data revealed elevated serum levels of tartrate resistant acid phosphatase (TRAP) without carbonic anhydrase II DNA mutation. A thoracolumbar spine X-ray showed, typical findings of ADO type II (ADO II; Albers-Schonberg disease), prominent vertebral endplates so called the 'rugger jersey spine'. Her older sister also showed same typical spine appearance. We report a case of ADO II with cavernous sinus thrombophlebitis and meningitis that was successfully treated with long-term antibiotics with right sphenoidotomy.
Abscess ; Acid Phosphatase ; Anti-Bacterial Agents ; Bone Density ; Bone Diseases ; Carbonic Anhydrase II ; Cavernous Sinus Thrombosis* ; DNA ; Female ; Humans ; Meningitis* ; Osteoclasts ; Osteopetrosis* ; Pelvis ; Sclerosis ; Siblings ; Skull Base ; Spine

We report 13 chromophobe renal cell carcinomas (10.8%) observed among 120 renal cell carcinomas in adults. The average age was 53 (range: 34-72) years old, and 6 were males and 7 females. The mean tumor size was 10 (range: 5-17) cm, mean nuclear grade 2.4, and mean Robson's stage was 1.9. There were two distinct histologic variants; typical variant (n=9) and eosinophilic variant (n=4). Both of them showed typical light microscopic features and positive reaction with Hale's colloidal iron and carbonic anhydrase II, a marker protein of intercalated cells of renal collecting ducts. A strong positive immunoreactivity for epithelial membrane antigen was noted in the cytoplasm in 12 of 13 tumors. Numerous microvesicles, 180~440 nm in diameter, were identified ultrastructurally. DNA aneuploidy was found in 3 out of 10 cases. Neither local recurrence nor metastasis have been identified during the following period of 4~144 (mean 48) months.
Adult ; Aneuploidy ; Carbonic Anhydrase II ; Carcinoma, Renal Cell* ; Colloids ; Cytoplasm ; DNA ; Eosinophils ; Female ; Humans ; Iron ; Male ; Mucin-1 ; Neoplasm Metastasis ; Recurrence

OBJECTIVETo study the clinicopathological significance of the expression of carbonic anhydrase (CA)II protein and mRNA in primary invasive ductal cancer (IDC) of human pancreas.

METHODSThe expression of CAII protein in 33 paired paraffin embedded IDC specimens of the pancreas and paired adjacent non-cancerous pancreatic tissues was detected by immunohistochemistry. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to examine the expression of CAII protein and mRNA level in 12 paired fresh IDC specimens of the pancreas and adjuvant non-cancerous pancreatic tissues. The relationship between the protein expression and clinicopathological features was analyzed.

RESULTSOverexpression of CAII protein was shown in 11 cases of pancreatic IDC tissues (33.3%, 11/33), which was much lower than that in paired non-cancerous pancreatic tissues (72.7%, t = 6.275, P = 0.000). The expression of CAII protein had no correlation with tumor position (χ² = 0.992, P = 0.319), differentiation (χ² = 0.866, P = 0.352), TNM stage (χ² = 1.210, P = 0.271) and Lymph node metastasis (χ² = 0.798, P = 0.372), but had bordering statistic sig with the prognosis of the patients (χ² = 3.233, P = 0.072). The median survival time in the patients with high expression of CAII protein was 540 days, while that in the patients with low expression was 320 days. The expression of CAII protein and mRNA was lower in IDC than that in paired non-cancerous pancreatic tissues detected by Western blot and RT-PCR respectively (t = 3.399, P = 0.006; t = 2.281, P = 0.043).

CONCLUSIONCAII is down regulated in pancreatic IDC and might be relative with the prognosis.

Carbonic Anhydrase II ; genetics ; metabolism ; Carcinoma, Pancreatic Ductal ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Pancreas ; metabolism ; Pancreatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics

OBJECTIVETo test expression of carbonic anhydrase II (Ca II) mRNA in osteoclasts which were applied with fluid shear stress.

METHODSThe bone marrow cells of Sprague-Dawley rats were cultured with the presence of 1,25-(OH)2D3 and dexamethasone. The osteoclast-like cells were identified by tartrate-resistant acid phosphatase(TRAP) staining and scanning electron microscope (SEM) observation, then purified with trypsin/ethylenediamine tetraacetic acid (EDTA). Different values and lasting time of steady fluid shear stress were exerted on the osteoclasts with parallel plate flow system. The Ca II expression of osteoclasts were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and nested polymerase chain reaction(PCR).

RESULTSThe levels of Ca II mRNA were down-regulated correspondingly with the increase of stress and time (P < 0.05).

CONCLUSIONIt's indicated that steady fluid shear stress within a certain range may down-regulate the expression of Ca II in osteoclasts.

Animals ; Bone Marrow Cells ; Carbonic Anhydrase II ; Cells, Cultured ; Osteoclasts ; Polymerase Chain Reaction ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical