Intravital Förster resonance energy transfer imaging reveals elevated [Ca2+]i and enhanced sympathetic tone in femoral arteries of angiotensin II-infused hypertensive biosensor mice

J Physiol. 2013 Nov 1;591(21):5321-36. doi: 10.1113/jphysiol.2013.257808. Epub 2013 Aug 27.

Abstract

Artery narrowing in hypertension can only result from structural remodelling of the artery, or increased smooth muscle contraction. The latter may occur with, or without, increases in [Ca(2+)]i. Here, we sought to measure, in living hypertensive mice, possible changes in artery dimensions and/or [Ca(2+)]i, and to determine some of the mechanisms involved. Ca(2+)/calmodulin biosensor (Förster resonance energy transfer-based) mice were made hypertensive by s.c. infusion of angiotensin II (Ang II, 400 ng kg(-1) min(-1), 2-3 weeks). Intravital fluorescence microscopy was used to determine [Ca(2+)]i and outer diameter of surgically exposed, intact femoral artery (FA) of anaesthetized mice. Active contractile FA 'tone' was calculated from the basal-state diameter and the passive (i.e. Ca(2+)-free) diameter (PD). Compared to saline control, FAs of Ang II-infused mice had (1) ∼21% higher active tone and (2) ∼78 nm higher smooth muscle [Ca(2+)]i, but (3) the same PDs. The local Ang II receptor (AT1R) blocker losartan had negligible effect on tone or [Ca(2+)]i in control FAs, but reduced the basal tone by ∼9% in Ang II FAs. Both i.v. hexamethonium and locally applied prazosin abolished the difference in FA tone and [Ca(2+)]i, suggesting a dominant role of sympathetic nerve activity (SNA). Changes in diameter and [Ca(2+)]i in response to locally applied phenylephrine, Ang II, arginine vasopressin, elevated [K(+)]o and acetylcholine were not altered. In summary, FAs of living Ang II hypertensive mice have higher [Ca(2+)]i, and are more constricted, due, primarily, to elevated SNA and some increased arterial AT1R activation. Evidence of altered artery reactivity or remodeling was not found.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin II / administration & dosage
  • Angiotensin II / toxicity
  • Animals
  • Antihypertensive Agents / pharmacology
  • Biosensing Techniques*
  • Calcium / metabolism*
  • Calcium Signaling*
  • Calmodulin / genetics
  • Calmodulin / metabolism
  • Femoral Artery / innervation
  • Femoral Artery / metabolism*
  • Femoral Artery / pathology
  • Femoral Artery / physiopathology
  • Fluorescence Resonance Energy Transfer*
  • Hexamethonium / pharmacology
  • Hypertension / chemically induced
  • Hypertension / metabolism*
  • Hypertension / physiopathology
  • Infusions, Intravenous
  • Losartan / pharmacology
  • Mice
  • Muscle, Smooth, Vascular / metabolism
  • Muscle, Smooth, Vascular / physiopathology
  • Prazosin / pharmacology
  • Sympathetic Nervous System / drug effects
  • Vasoconstriction*

Substances

  • Antihypertensive Agents
  • Calmodulin
  • Angiotensin II
  • Hexamethonium
  • Losartan
  • Calcium
  • Prazosin