Carbonic anhydrase II mRNA is induced in rabbit kidney cortex during chronic metabolic acidosis

George J. Schwartz, Cornelia A. Winkler, Beth J. Zavilowitz, Thaddeus Bargiello

Research output: Contribution to journalArticle

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Abstract

Carbonic anhydrase II (CA II), the predominant isoform of carbonic anhydrase in the kidney, is believed to be localized primarily in the cytoplasm of proximal tubule and collecting duct intercalated cells. Carbonic anhydrase facilitates H+ secretion by catalyzing the formation of HCO3- from OH- in the presence of CO2. We have shown that renal cortical CA II activity is stimulated during 4-6 days of chronic metabolic acidosis [L. P. Brion, B. J. Zavilowitz, O. Rosen, and G. J. Schwartz. Am. J. Physiol. 261 (Regulatory Integrative Comp. Physiol. 30): R1204-R1213, 1991]. The purpose of these studies was to examine under similar conditions the regulation of CA II mRNA. We obtained a major portion of the rabbit CA II cDNA by reverse transcription of total RNA from rabbit kidney followed by amplification using oligonucleotide primers prepared from conserved areas in the coding regions of human, mouse, and chick CAII cDNAs in a polymerase chain reaction (RT-PCR). The 696-bp RT-PCR product was sequenced and found to be 71-86% homologous to CA II cDNAs from the other three species. The deduced amino acid sequence agreed closely (>97%) with a previous Edman analysis of rabbit erythrocyte CA II. Northern analysis showed expression of a ∼ 1.4 kb RNA, with cortex > outer medulla > inner medulla. Steady-state mRNA expression from kidney cortex of acid-treated rabbits was about twice that from controls, when normalized to the expression of β-actin or malate dehydrogenase. The stimulation of CA II mRNA was greater after 3 days than after 5-6 days of acid treatment. Thus, during chronic metabolic acidosis, there is an induction of CA II mRNA, which precedes the increase in activity, indicating a possible role for this enzyme in the renal adaptation to this pathological state.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume265
Issue number6 34-6
StatePublished - Dec 1993
Externally publishedYes

Fingerprint

Carbonic Anhydrase II
Kidney Cortex
Acidosis
Rabbits
Messenger RNA
Kidney
Carbonic Anhydrases
Complementary DNA
Polymerase Chain Reaction
RNA
Malate Dehydrogenase
Acids
DNA Primers
Reverse Transcription
Actins
Amino Acid Sequence
Protein Isoforms
Cytoplasm
Erythrocytes

Keywords

  • β-actin
  • Ammonium chloride
  • Chick liver
  • Chick muscle
  • Malate dehydrogenase
  • Mouse erythroleukemic cells
  • Nucleotide sequence
  • Outer medulla
  • Polymerase chain reaction
  • Reverse transcription

ASJC Scopus subject areas

  • Physiology

Cite this

Carbonic anhydrase II mRNA is induced in rabbit kidney cortex during chronic metabolic acidosis. / Schwartz, George J.; Winkler, Cornelia A.; Zavilowitz, Beth J.; Bargiello, Thaddeus.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 265, No. 6 34-6, 12.1993.

Research output: Contribution to journalArticle

Schwartz, George J. ; Winkler, Cornelia A. ; Zavilowitz, Beth J. ; Bargiello, Thaddeus. / Carbonic anhydrase II mRNA is induced in rabbit kidney cortex during chronic metabolic acidosis. In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology. 1993 ; Vol. 265, No. 6 34-6.
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abstract = "Carbonic anhydrase II (CA II), the predominant isoform of carbonic anhydrase in the kidney, is believed to be localized primarily in the cytoplasm of proximal tubule and collecting duct intercalated cells. Carbonic anhydrase facilitates H+ secretion by catalyzing the formation of HCO3- from OH- in the presence of CO2. We have shown that renal cortical CA II activity is stimulated during 4-6 days of chronic metabolic acidosis [L. P. Brion, B. J. Zavilowitz, O. Rosen, and G. J. Schwartz. Am. J. Physiol. 261 (Regulatory Integrative Comp. Physiol. 30): R1204-R1213, 1991]. The purpose of these studies was to examine under similar conditions the regulation of CA II mRNA. We obtained a major portion of the rabbit CA II cDNA by reverse transcription of total RNA from rabbit kidney followed by amplification using oligonucleotide primers prepared from conserved areas in the coding regions of human, mouse, and chick CAII cDNAs in a polymerase chain reaction (RT-PCR). The 696-bp RT-PCR product was sequenced and found to be 71-86{\%} homologous to CA II cDNAs from the other three species. The deduced amino acid sequence agreed closely (>97{\%}) with a previous Edman analysis of rabbit erythrocyte CA II. Northern analysis showed expression of a ∼ 1.4 kb RNA, with cortex > outer medulla > inner medulla. Steady-state mRNA expression from kidney cortex of acid-treated rabbits was about twice that from controls, when normalized to the expression of β-actin or malate dehydrogenase. The stimulation of CA II mRNA was greater after 3 days than after 5-6 days of acid treatment. Thus, during chronic metabolic acidosis, there is an induction of CA II mRNA, which precedes the increase in activity, indicating a possible role for this enzyme in the renal adaptation to this pathological state.",
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AU - Zavilowitz, Beth J.

AU - Bargiello, Thaddeus

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N2 - Carbonic anhydrase II (CA II), the predominant isoform of carbonic anhydrase in the kidney, is believed to be localized primarily in the cytoplasm of proximal tubule and collecting duct intercalated cells. Carbonic anhydrase facilitates H+ secretion by catalyzing the formation of HCO3- from OH- in the presence of CO2. We have shown that renal cortical CA II activity is stimulated during 4-6 days of chronic metabolic acidosis [L. P. Brion, B. J. Zavilowitz, O. Rosen, and G. J. Schwartz. Am. J. Physiol. 261 (Regulatory Integrative Comp. Physiol. 30): R1204-R1213, 1991]. The purpose of these studies was to examine under similar conditions the regulation of CA II mRNA. We obtained a major portion of the rabbit CA II cDNA by reverse transcription of total RNA from rabbit kidney followed by amplification using oligonucleotide primers prepared from conserved areas in the coding regions of human, mouse, and chick CAII cDNAs in a polymerase chain reaction (RT-PCR). The 696-bp RT-PCR product was sequenced and found to be 71-86% homologous to CA II cDNAs from the other three species. The deduced amino acid sequence agreed closely (>97%) with a previous Edman analysis of rabbit erythrocyte CA II. Northern analysis showed expression of a ∼ 1.4 kb RNA, with cortex > outer medulla > inner medulla. Steady-state mRNA expression from kidney cortex of acid-treated rabbits was about twice that from controls, when normalized to the expression of β-actin or malate dehydrogenase. The stimulation of CA II mRNA was greater after 3 days than after 5-6 days of acid treatment. Thus, during chronic metabolic acidosis, there is an induction of CA II mRNA, which precedes the increase in activity, indicating a possible role for this enzyme in the renal adaptation to this pathological state.

AB - Carbonic anhydrase II (CA II), the predominant isoform of carbonic anhydrase in the kidney, is believed to be localized primarily in the cytoplasm of proximal tubule and collecting duct intercalated cells. Carbonic anhydrase facilitates H+ secretion by catalyzing the formation of HCO3- from OH- in the presence of CO2. We have shown that renal cortical CA II activity is stimulated during 4-6 days of chronic metabolic acidosis [L. P. Brion, B. J. Zavilowitz, O. Rosen, and G. J. Schwartz. Am. J. Physiol. 261 (Regulatory Integrative Comp. Physiol. 30): R1204-R1213, 1991]. The purpose of these studies was to examine under similar conditions the regulation of CA II mRNA. We obtained a major portion of the rabbit CA II cDNA by reverse transcription of total RNA from rabbit kidney followed by amplification using oligonucleotide primers prepared from conserved areas in the coding regions of human, mouse, and chick CAII cDNAs in a polymerase chain reaction (RT-PCR). The 696-bp RT-PCR product was sequenced and found to be 71-86% homologous to CA II cDNAs from the other three species. The deduced amino acid sequence agreed closely (>97%) with a previous Edman analysis of rabbit erythrocyte CA II. Northern analysis showed expression of a ∼ 1.4 kb RNA, with cortex > outer medulla > inner medulla. Steady-state mRNA expression from kidney cortex of acid-treated rabbits was about twice that from controls, when normalized to the expression of β-actin or malate dehydrogenase. The stimulation of CA II mRNA was greater after 3 days than after 5-6 days of acid treatment. Thus, during chronic metabolic acidosis, there is an induction of CA II mRNA, which precedes the increase in activity, indicating a possible role for this enzyme in the renal adaptation to this pathological state.

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KW - Chick muscle

KW - Malate dehydrogenase

KW - Mouse erythroleukemic cells

KW - Nucleotide sequence

KW - Outer medulla

KW - Polymerase chain reaction

KW - Reverse transcription

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