Expression of the cysteine proteinase inhibitor cystatin C gene in rat heart: use of digoxigenin-labeled probes generated by polymerase chain reaction directly for in situ and northern blot hybridizations.
Cystatins represent a widely distributed superfamily of cysteine proteinase inhibitory proteins. We investigated the expression of the cystatin C gene, belonging to the family 2 of cystatins, in the hearts of female rats. Using a highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) we have detected cystatin C mRNA in the ventricule and atrium, as well as in liver and submandibular gland. A digoxigenin-labeled cystatin C probe, generated by PCR, hybridized to a single mRNA species of about 700 nucleotides on Northern blots. Northern blot hybridizations established that neither an acute inflammation produced by injection of turpentine nor administration of the beta-adrenergic agonist isoproterenol had an effect on the level of cystatin C mRNA in the heart. In situ hybridizations with digoxigenin-labeled probe localized the expression of the cystatin C gene to cardiac muscle fibers but not to other cardiac cellular elements. Cystatin C may be released by cardiac muscle fibers under physiological and pathological conditions and may modify inflammatory and necrobiotic processes.
Barka
T
T
Department of Cell Biology/Anatomy, Mount Sinai School of Medicine, City University of New York, NY 10029-6574.
van der Noen
H
H
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D055316:Cystatin C; D015891:Cystatins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D015853:Cysteine Proteinase Inhibitors / Q000032:analysis / Q000235:genetics*; D004076:Digoxigenin; D005260:Female; D015870:Gene Expression / Q000235:genetics*; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011943:Receptors, Adrenergic, beta / Q000502:physiology
C525032:Cst3 protein, rat; D055316:Cystatin C; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D012333:RNA, Messenger; D011943:Receptors, Adrenergic, beta; D004076:Digoxigenin