Kallikrein messenger RNA in rat arteries and veins

G. M. Saed, O. A. Carretero, R. J. MacDonald, A. G. Scicli

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Glandular kallikrein (EC 3.4.21.8) belongs to a subgroup of serine proteases coded by a multigene family. A kininogenase resembling glandular kallikrein has been identified in vascular tissue; however, it is not clear whether it is synthesized by vascular tissue or taken up from plasma. To determine the potential for kallikrein synthesis in vascular tissues, we tested whether messenger RNA (mRNA) for glandular kallikrein is present in rat arteries and veins. Poly(A+) RNA was isolated from pools of arteries or veins (n = 3, 30 rats each). Poly(A+) RNA from the kidney and liver was used as a positive and negative control, respectively. As a probe, we used rat pancreatic kallikrein 32P-labeled complementary DNA, which recognizes mRNA of the entire rat kallikrein family. Slot-blot analysis indicated htat kallikrein mRNA was present in mRNA from the arteries, veins, and kidney but not from the liver. Poly(A+) RNA from arteries and veins contained ~1% as much kallikrein mRNA as that from the kidney. To confirm the slot-blot results and determine whether the mRNA for true glandular kallikrein was present in vascular tissue, we employed a polymerase chain reaction assay, first using primers specific for the entire kallikrein family (which amplify a 430-bp fragment) and then using primers specific for true glandular kallikrein mRNA (which amplify a 370-bp fragment). After the polymerase chain reaction assay, both arteries and veins showed fragments of these sizes when tested with rat kallikrein complementary DNA probe, thus confirming the presence of glandular kallikrein mRNA. Similar results were obtained when the polymerase chain reaction assay was applied to mRNA isolated from vascular smooth muscle cells in culture and from the kidney. No signal was obtained with liver mRNA. We concluded that kallikrein is synthesized by the vascular wall, possibly by smooth muscle cells. The presence of locally synthesized kallikrein indicates that the vascular kallikrein-kinin system may play a role in the regulation of vascular tone.

Original languageEnglish (US)
Pages (from-to)510-516
Number of pages7
JournalCirculation Research
Volume67
Issue number2
StatePublished - 1990

Fingerprint

Kallikreins
Veins
Arteries
Tissue Kallikreins
Messenger RNA
Blood Vessels
Kidney
Polymerase Chain Reaction
Smooth Muscle Myocytes
Liver
Complementary DNA
Kallikrein-Kinin System
DNA Probes
Serine Proteases
Multigene Family
Vascular Smooth Muscle

Keywords

  • Kallikrein gene
  • Kallikrein-kinin system
  • Polymerase chain reaction
  • Vascular smooth muscle cells
  • Vascular tissue

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Saed, G. M., Carretero, O. A., MacDonald, R. J., & Scicli, A. G. (1990). Kallikrein messenger RNA in rat arteries and veins. Circulation Research, 67(2), 510-516.

Kallikrein messenger RNA in rat arteries and veins. / Saed, G. M.; Carretero, O. A.; MacDonald, R. J.; Scicli, A. G.

In: Circulation Research, Vol. 67, No. 2, 1990, p. 510-516.

Research output: Contribution to journalArticle

Saed, GM, Carretero, OA, MacDonald, RJ & Scicli, AG 1990, 'Kallikrein messenger RNA in rat arteries and veins', Circulation Research, vol. 67, no. 2, pp. 510-516.
Saed GM, Carretero OA, MacDonald RJ, Scicli AG. Kallikrein messenger RNA in rat arteries and veins. Circulation Research. 1990;67(2):510-516.
Saed, G. M. ; Carretero, O. A. ; MacDonald, R. J. ; Scicli, A. G. / Kallikrein messenger RNA in rat arteries and veins. In: Circulation Research. 1990 ; Vol. 67, No. 2. pp. 510-516.
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AB - Glandular kallikrein (EC 3.4.21.8) belongs to a subgroup of serine proteases coded by a multigene family. A kininogenase resembling glandular kallikrein has been identified in vascular tissue; however, it is not clear whether it is synthesized by vascular tissue or taken up from plasma. To determine the potential for kallikrein synthesis in vascular tissues, we tested whether messenger RNA (mRNA) for glandular kallikrein is present in rat arteries and veins. Poly(A+) RNA was isolated from pools of arteries or veins (n = 3, 30 rats each). Poly(A+) RNA from the kidney and liver was used as a positive and negative control, respectively. As a probe, we used rat pancreatic kallikrein 32P-labeled complementary DNA, which recognizes mRNA of the entire rat kallikrein family. Slot-blot analysis indicated htat kallikrein mRNA was present in mRNA from the arteries, veins, and kidney but not from the liver. Poly(A+) RNA from arteries and veins contained ~1% as much kallikrein mRNA as that from the kidney. To confirm the slot-blot results and determine whether the mRNA for true glandular kallikrein was present in vascular tissue, we employed a polymerase chain reaction assay, first using primers specific for the entire kallikrein family (which amplify a 430-bp fragment) and then using primers specific for true glandular kallikrein mRNA (which amplify a 370-bp fragment). After the polymerase chain reaction assay, both arteries and veins showed fragments of these sizes when tested with rat kallikrein complementary DNA probe, thus confirming the presence of glandular kallikrein mRNA. Similar results were obtained when the polymerase chain reaction assay was applied to mRNA isolated from vascular smooth muscle cells in culture and from the kidney. No signal was obtained with liver mRNA. We concluded that kallikrein is synthesized by the vascular wall, possibly by smooth muscle cells. The presence of locally synthesized kallikrein indicates that the vascular kallikrein-kinin system may play a role in the regulation of vascular tone.

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