Use the Laser Speckle Imaging System to measure blood flow in cerebral arteries via a cranial window in anesthetized mice, to corroborate an in vivo physiological role for pS1928 upon HG.

Martín-Aragón Baudel M, Flores-Tamez V A, Hong J, et al. Spatiotemporal control of vascular CaV1. 2 by α1c S1928 phosphorylation[J]. Circulation Research, 2022, 131(12): 1018-1033.

Journal: Circulation Research

Background and Customer Requirements:

• 1) L-type Ca_V1.2 channels undergo cooperative gating to regulate cell function, although mechanisms are unclear.
• 2) This study aim to test the hypothesis that phosphorylation of the Ca_V1.2 pore-forming subunit α-1C at S1928 mediates vascular Ca_V1.2 cooperativity during diabetic hyperglycemia.

Demands of the Research:

The main purpose of this study was to investigate the effect of phosphorylation of the Ca_V1.2 pore-forming subunit α-1C at S1928 mediates vascular Ca_V1.2 cooperativity. And it is crucial to get the blood flow to assess the effect.

What We Offer:

Product: RFLSI Laser Speckle Imaging System

Clear to see that representative pseudo-colored blood flow images of WT and S1928A cerebral pial arteries through a cranial window exposed to 10 mmol/L D-glu, 20 mmol/L D-glu or 0 Ca²⁺ vasodilatory mix, which confirms that pS1928 is necessary for increased arterial myocyte Ca²⁺ and contractility upon HG.

Results and Effects:

• 1) Ca_V1.2 activity and cooperative gating increase in arterial myocytes from patients with type 2 diabetes and type 1 diabetic mice, and in wild-type mouse arterial myocytes after elevating extracellular glucose，which was prevented in wild-type cells pre-exposed to a PKA inhibitor or cells from knock-in S1928A but not S1700A mice.
• 2) α-1_C clustering at the surface membrane of wild-type, but not wild-type cells pre-exposed to PKA or P2Y11 inhibitors and S1928A arterial myocytes, was elevated upon hyperglycemia and diabetes.
• 3) Ca_V1.2 spatial and gating remodeling correlated with enhanced arterial myocyte Ca²⁺ influx and contractility and in vivo reduction in arterial diameter and blood flow upon hyperglycemia and diabetes in wild-type but not S1928A cells/mice.
• 4) The results may lay the foundation for developing therapeutics to correct Ca_V1.2 and arterial function during diabetic hyperglycemia.