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All nine transmembrane adenylyl cyclase isoforms are temporally expressed in rat articular chondrocytes
Author(s) -
Memon Ismail,
Khan Khalid,
Siddiqui Sammer,
Perveen Siddiqa,
Ishaq Muhammad
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.492.6
Subject(s) - adenylyl cyclase , gene isoform , adcy9 , transmembrane protein , adcy10 , microbiology and biotechnology , chemistry , biology , gene , signal transduction , biochemistry , receptor
Identification of transmembrane adenylyl cyclase (tm‐AC) isoforms and their temporal expression in articular chondrocytes in rats was investigated using RT‐PCR and immunohistochemistry techniques. For RT‐PCR, cDNA was made from RNA extracted from articular chondrocytes. PCR was run with primers specific for tm‐AC isoforms. PCR bands were normalized and quantified with β actin, and densitometry analysis was carried out. For immunohistochemistry, articular cartilage sections were incubated with antibodies specific for tm‐AC isoforms and peroxidase activity was demonstrated by Diaminobenzidine and H 2 O 2 in Tris‐buffered saline. All tm‐AC isoforms were expressed in chondrocytes from all age groups examined. Each isoform was differentially expressed in developing and adult articular chondrocytes. Generally, expression of AC isoforms was observed to increase with age, but increase was not uniform for all tm‐AC isoforms. Expression of tm‐AC isoforms along with other signaling molecules known to be present in articular chondrocytes indicate complicated and multifactorial signaling cascades involved in the development and homeostasis of articular cartilage. This research work was funded by a seed money grant from the FHS‐Research Committee of the Aga Khan University Grant Funding Source: AAA