TY - JOUR
T1 - Combined intravital microscopy and contrast-enhanced ultrasonography of the mouse hindlimb to study insulin-induced vasodilation and muscle perfusion
AU - Turaihi, Alexander H.
AU - van Poelgeest, Erik M.
AU - van Hinsbergh, Victor W M
AU - Serné, Erik H.
AU - Smulders, Yvo M.
AU - Eringa, Etto C.
PY - 2017/3/20
Y1 - 2017/3/20
N2 - It has been demonstrated that insulin’s vascular actions contribute to regulation of insulin sensitivity. Insulin’s effects on muscle perfusion regulate postprandial delivery of nutrients and hormones to insulin-sensitive tissues. We here describe a technique for combining intravital microscopy (IVM) and contrast-enhanced ultrasonography (CEUS) of the adductor compartment of the mouse hindlimb to simultaneously visualize muscle resistance arteries and perfusion of the microcirculation in vivo. Simultaneously assessing insulin’s effect at multiple levels of the vascular tree is important to study relationships between insulin’s multiple vasoactive effects and muscle perfusion. Experiments in this study were performed in mice. First, the tail vein cannula is inserted for the infusion of anesthesia, vasoactive compounds and ultrasound contrast agent (lipid-encapsulated microbubbles). Second, a small incision is made in the groin area to expose the arterial tree of the adductor muscle compartment. The ultrasound probe is then positioned at the contralateral upper hindlimb to view the muscles in cross-section. To assess baseline parameters, the arterial diameter is assessed and microbubbles are subsequently infused at a constant rate to estimate muscle blood flow and microvascular blood volume (MBV). When applied before and during a hyperinsulinemic-euglycemic clamp, combined IVM and CEUS allow assessment of insulin-induced changes of arterial diameter, microvascular muscle perfusion and whole-body insulin sensitivity. Moreover, the temporal relationship between responses of the microcirculation and the resistance arteries to insulin can be quantified. It is also possible to follow-up the mice longitudinally in time, making it a valuable tool to study changes in vascular and whole-body insulin sensitivity.
AB - It has been demonstrated that insulin’s vascular actions contribute to regulation of insulin sensitivity. Insulin’s effects on muscle perfusion regulate postprandial delivery of nutrients and hormones to insulin-sensitive tissues. We here describe a technique for combining intravital microscopy (IVM) and contrast-enhanced ultrasonography (CEUS) of the adductor compartment of the mouse hindlimb to simultaneously visualize muscle resistance arteries and perfusion of the microcirculation in vivo. Simultaneously assessing insulin’s effect at multiple levels of the vascular tree is important to study relationships between insulin’s multiple vasoactive effects and muscle perfusion. Experiments in this study were performed in mice. First, the tail vein cannula is inserted for the infusion of anesthesia, vasoactive compounds and ultrasound contrast agent (lipid-encapsulated microbubbles). Second, a small incision is made in the groin area to expose the arterial tree of the adductor muscle compartment. The ultrasound probe is then positioned at the contralateral upper hindlimb to view the muscles in cross-section. To assess baseline parameters, the arterial diameter is assessed and microbubbles are subsequently infused at a constant rate to estimate muscle blood flow and microvascular blood volume (MBV). When applied before and during a hyperinsulinemic-euglycemic clamp, combined IVM and CEUS allow assessment of insulin-induced changes of arterial diameter, microvascular muscle perfusion and whole-body insulin sensitivity. Moreover, the temporal relationship between responses of the microcirculation and the resistance arteries to insulin can be quantified. It is also possible to follow-up the mice longitudinally in time, making it a valuable tool to study changes in vascular and whole-body insulin sensitivity.
KW - Contrast-enhanced ultrasonography
KW - Endothelium
KW - Insulin sensitivity
KW - Intravital microscopy
KW - Issue 121
KW - Medicine
KW - Microsurgery
KW - Vascular responsiveness
UR - http://www.scopus.com/inward/record.url?scp=85017144967&partnerID=8YFLogxK
U2 - https://doi.org/10.3791/54912
DO - https://doi.org/10.3791/54912
M3 - Article
C2 - 28362362
SN - 1940-087X
VL - 2017
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 121
M1 - e54912
ER -