Evidence by signal peptide trap technology for the expression of carbonic anhydrase 6 in rat incisor enamel organs

During screening of a rat incisor enamel organ cDNA library by signal peptide trap technology, we identified a DNA fragment matching a predicted translation sequence for rat carbonic anhydrase 6 (CA6). This result was unexpected because CA6, to date, has been associated primarily with secretions fro...

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Bibliographic Details
Published in:European journal of oral sciences 2006-05, Vol.114 (s1), p.147-153
Main Authors: Smith, Charles E., Nanci, Antonio, Moffatt, Pierre
Format: Article
Language:English
Subjects:
Amelogenesis - genetics
Animals
Apatites - chemistry
bicarbonate
CA6
carbonic anhydrase
Carbonic Anhydrases - analysis
Carbonic Anhydrases - genetics
Cloning, Molecular
Crystallography
Dental Enamel - enzymology
Dental Enamel - secretion
Dentistry
DNA, Complementary - genetics
enamel formation
Enamel Organ - enzymology
Enamel Organ - secretion
Incisor
Molecular Sequence Data
Protein Biosynthesis - genetics
Protein Sorting Signals - genetics
Protein Subunits - analysis
Protein Subunits - genetics
Rats
Reverse Transcriptase Polymerase Chain Reaction
Salivary Glands - enzymology
Sequence Analysis, Protein
tooth
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Summary:During screening of a rat incisor enamel organ cDNA library by signal peptide trap technology, we identified a DNA fragment matching a predicted translation sequence for rat carbonic anhydrase 6 (CA6). This result was unexpected because CA6, to date, has been associated primarily with secretions from glandular tissues. To further characterize this observation, reverse transcription–polymerase chain reaction (RT–PCR) amplifications were carried out on total RNA extracted from freeze‐dried secretory and maturation‐stage rat incisor enamel organs. A cDNA fragment of the expected size was detected in control samples from rat salivary glands as well as within maturation‐stage enamel organ samples. This CA6 RT–PCR fragment was further cloned and sequenced and found to match the nucleotide sequence 770–1079 from clone XM_216584 of GenBank. Northern blot analyses with the rat CA6 cDNA fragment confirmed its expression relative to maturation‐stage enamel organ samples. It is at present unclear whether the CA6 expressed by enamel organ cells is secreted into the enamel layer or into the intercellular spaces of the enamel organ itself to assist in neutralizing excess protons arising from the growth of apatite crystals during the maturation stage of amelogenesis.
ISSN:0909-8836
1600-0722
DOI:10.1111/j.1600-0722.2006.00273.x