DOI: 10.5061/dryad.k3j9kd5dz
title: Impaired intracellular Ca2+ signaling contributes to age-related cerebral small vessel disease in Col4a1 mutant mice

description: Humans and mice with mutations in COL4A1 and COL4A2 manifest hallmarks of
 cerebral small vessel disease (cSVD). Mice with a missense mutation in
 Col4a1 at amino acid 1344 (Col4a1+/G1344D) exhibit age-dependent
 intracerebral hemorrhage (ICH) and brain lesions. Here we report that this
 pathology was associated with the loss of myogenic vasoconstriction, an
 intrinsic vascular response essential for the autoregulation of cerebral
 blood flow. Electrophysiological analyses showed that the loss of myogenic
 constriction resulted from blunted pressure-induced smooth muscle cell
 (SMC) membrane depolarization. Further, we found that dysregulation of
 membrane potential was associated with impaired Ca2+-dependent activation
 of large-conductance Ca2+-activated K+ (BK) and transient receptor
 potential melastatin 4 (TRPM4) cation channels linked to disruptions in
 sarcoplasmic reticulum (SR) Ca2+ signaling.  Treating
 Col4a1+/G1344D mice with 4-phenylbutyrate, a compound that promotes the
 trafficking of misfolded proteins and alleviates SR stress, restored
 SR Ca2+ signaling and BK and TRPM4 channel activity, prevented
 loss of myogenic tone, and reduced ICH. We conclude that alterations in SR
 Ca2+ handling that impair membrane potential regulating ion channel
 activity result in dysregulation of SMC membrane potential and loss of
 myogenic tone contributing to age-related cSVD in Col4a1+/G1344D mice.
description: # Data set for "Impaired intracellular Ca2+ signaling contributes to
 age-related cerebral small vessel disease in Col4a1 mutant mice" ---
 This dataset contains all the processed data published in "Impaired
 intracellular Ca2+ signaling contributes to age-related cerebral small
 vessel disease in Col4a1 mutant mice". ## Description of the data and
 file structure Figure_1_Processed_Data * 1C: Percentage of brain area with
 Prussian blue staining in brain sections from 3 M-old Col4a1+/+ and
 Col4a1+/G1344D mice. * 1E: Percentage of brain area with Prussian blue
 staining in brain sections from 12 M-old Col4a1+/+ and Col4a1+/G1344D
 mice. * 1G: The number of brain lesions detected with high-field
 susceptibility-weighted magnetic resonance imaging of 12 M-old Col4a1+/+
 and Col4a1+/G1344D mice. * 1H: Total volume of brain lesions detected with
 high-field susceptibility-weighted magnetic resonance imaging of 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. * 1I: Localization of brain lesions
 detected with high-field susceptibility-weighted magnetic resonance
 imaging of 12 M-old Col4a1+/+ and Col4a1+/G1344D mice.
 Figure_2_Processed_Data * 2A: Inner diameter of isolated cerebral arteries
 from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice show the response to
 increased pressure in the presence (active) and absence (passive) of
 extracellular Ca2+. * 2B: Summary data of myogenic tone as a function of
 intraluminal pressure in cerebral arteries from 3 M-old Col4a1+/+ and
 Col4a1+/G1344D mice. * 2C: Inner diameter of isolated cerebral arteries
 from 12 M-old Col4a1+/+ and Col4a1+/G1344D mice show the response to
 increased pressure in the presence (active) and absence (passive) of
 extracellular Ca2+. * 2D: Summary data of myogenic tone as a function of
 intraluminal pressure in cerebral arteries from 12 M-old Col4a1+/+ and
 Col4a1+/G1344D mice. * 2E: Intracellular membrane potential (mV)
 recordings of smooth muscle cells in pressurized cerebral arteries
 isolated from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice at 20 mmHg and 80
 mmHg intraluminal pressure. * 2F: Summary data showing membrane potential
 of smooth muscle cells in pressurized cerebral arteries from 3 M-old
 Col4a1+/+ and Col4a1+/G1344D mice at 20 mmHg and 80 mmHg intraluminal
 pressure. * 2G: Intracellular membrane potential (mV) recordings of smooth
 muscle cells in pressurized cerebral arteries isolated from 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice at 20 mmHg and 80 mmHg intraluminal
 pressure. * 2H: Summary data showing membrane potential of smooth muscle
 cells in pressurized cerebral arteries from 12 M-old Col4a1+/+ and
 Col4a1+/G1344D mice at 20 mmHg and 80 mmHg intraluminal pressure.
 Figure_3_Processed_Data * 3B: Frequency of spontaneous transient outward
 currents in freshly isolated cerebral artery smooth muscle cells from 3
 M-old Col4a1+/+ and Col4a1+/G1344D mice over a range of membrane
 potentials (-60 to 0 mV). * 3C: Amplitude of spontaneous transient outward
 currents in freshly isolated cerebral artery smooth muscle cells from 3
 M-old Col4a1+/+ and Col4a1+/G1344D mice over a range of membrane
 potentials (-60 to 0 mV). * 3E: Frequency of spontaneous transient outward
 currents in freshly isolated cerebral artery smooth muscle cells from 12
 M-old Col4a1+/+ and Col4a1+/G1344D mice over a range of membrane
 potentials (-60 to 0 mV). * 3F: Amplitude of spontaneous transient outward
 currents in freshly isolated cerebral artery smooth muscle cells from 12
 M-old Col4a1+/+ and Col4a1+/G1344D mice over a range of membrane
 potentials (-60 to 0 mV). * 3G: I-V relationship of whole-cell patch-clamp
 TRPM4 current recordings during voltage ramps (-100 to 100 mV) in cerebral
 artery smooth muscle cells from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice.
 * 3H: Whole-cell TRPM4 current density at +100 mV in cerebral artery
 smooth muscle cells from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice. * 3I:
 I-V relationship of whole-cell patch-clamp TRPM4 current recordings during
 voltage ramps (-100 to 100 mV) in cerebral artery smooth muscle cells from
 12 M-old Col4a1+/+ and Col4a1+/G1344D mice. * 3J: Whole-cell TRPM4 current
 density at +100 mV in cerebral artery smooth muscle cells from 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. * 3L: Transient inward cation current
 activity in cerebral artery smooth muscle cells isolated from 3 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. * 3N: Transient inward cation current
 activity in cerebral artery smooth muscle cells isolated from 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. Figure_4_Processed_Data * 4B:
 Representative Ca2+ spark events in cerebral artery smooth muscle cells
 isolated from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice presented as
 changes in fractional fluorescence. * 4C: Ca2+ spark frequency, amplitude,
 duration (t1/2), rise time, and decay (t1/2) in cerebral artery smooth
 muscle cells isolated from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice. *
 4E: Representative Ca2+ spark events in cerebral artery smooth muscle
 cells isolated from 12 M-old Col4a1+/+ and Col4a1+/G1344D mice presented
 as changes in fractional fluorescence. * 4F: Ca2+ spark frequency,
 amplitude, duration (t1/2), rise time, and decay (t1/2) in cerebral artery
 smooth muscle cells isolated from 12 M-old Col4a1+/+ and Col4a1+/G1344D
 mice. * 4G: Traces of whole-cell fractional fluorescence changes in
 response to caffeine in Fluo-4-AM loaded cerebral artery smooth muscle
 cells from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice. * 4H: Peak
 whole-cell fractional fluorescence changes in response to caffeine in
 Fluo-4-AM loaded cerebral artery smooth muscle cells from 3 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. * 4I: Traces of whole-cell fractional
 fluorescence changes in response to caffeine in Fluo-4-AM loaded cerebral
 artery smooth muscle cells from 12 M-old Col4a1+/+ and Col4a1+/G1344D
 mice. * 4J: Peak whole-cell fractional fluorescence changes in response to
 caffeine in Fluo-4-AM loaded cerebral artery smooth muscle cells from 12
 M-old Col4a1+/+ and Col4a1+/G1344D mice. * 4K: Traces of whole-cell
 fractional fluorescence changes in response to U46619 in Fluo-4-AM loaded
 cerebral artery smooth muscle cells from 3 M-old Col4a1+/+ and
 Col4a1+/G1344D mice. * 4L: Peak whole-cell fractional fluorescence changes
 in response to U46619 in Fluo-4-AM loaded cerebral artery smooth muscle
 cells from 3 M-old Col4a1+/+ and Col4a1+/G1344D mice. * 4M: Traces of
 whole-cell fractional fluorescence changes in response to U46619 in
 Fluo-4-AM loaded cerebral artery smooth muscle cells from 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice. * 4N: Peak whole-cell fractional
 fluorescence changes in response to U46619 in Fluo-4-AM loaded cerebral
 artery smooth muscle cells from 12 M-old Col4a1+/+ and Col4a1+/G1344D
 mice. Figure_5_Processed_Data * 5A: Traces showing whole-cell fractional
 fluorescence changes in response to caffeine in Fluo-4-AM loaded cerebral
 artery smooth muscle cells from 4PBA treated 12 M-old Col4a1+/+ and
 Col4a1+/G1344D and untreated Col4a1+/G1344D mice. * 5B: Peak whole-cell
 fractional fluorescence changes in response to caffeine in Fluo-4-AM
 loaded cerebral artery smooth muscle cells from 4PBA treated 12 M-old
 Col4a1+/+ and Col4a1+/G1344D and untreated Col4a1+/G1344D mice. * 5C:
 Traces showing whole-cell fractional fluorescence changes in response to
 U46619 in Fluo-4-AM loaded cerebral artery smooth muscle cells from 4PBA
 treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and untreated Col4a1+/G1344D
 mice. * 5D: Peak whole-cell fractional fluorescence changes in response to
 U46619 in Fluo-4-AM loaded cerebral artery smooth muscle cells from 4PBA
 treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and untreated Col4a1+/G1344D
 mice. * 5F: Transient inward cation current activity in cerebral artery
 smooth muscle cells isolated from 4PBA treated 12 M-old Col4a1+/+ and
 Col4a1+/G1344D and untreated Col4a1+/G1344D mice. * 5G: Inner diameter of
 isolated cerebral arteries from 4PBA treated 12 M-old Col4a1+/+ and
 Col4a1+/G1344D and untreated Col4a1+/G1344D mice show the response to
 increased pressure in the presence (active) and absence (passive) of
 extracellular Ca2+. * 5H: Summary data of myogenic tone as a function of
 intraluminal pressure in cerebral arteries from 4PBA treated 12 M-old
 Col4a1+/+ and Col4a1+/G1344D and untreated Col4a1+/G1344D mice. * 5J:
 Percentage of brain area with Prussian blue staining in brain sections
 from 4PBA treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and untreated
 Col4a1+/G1344D mice. Supplemental_Figure_1_Processed_Data * S1A: Summary
 data of passive lumen diameter as a function of intraluminal pressure in
 cerebral arteries from 12 M-old Col4a1+/+ and Col4a1+/G1344D mice.
 Supplemental_Figure_2_Processed_Data * S2A: Traces of
 4-aminopyridine-sensitive KV currents in freshly isolated cerebral artery
 smooth muscle cells from 12 M-old Col4a1+/+ and Col4a1+/G1344D mice. *
 S2B: KV currents normalized to cell capacitance in freshly isolated
 cerebral artery smooth muscle cells from 12 M-old Col4a1+/+ and
 Col4a1+/G1344D mice. Supplemental_Figure_3_Processed_Data * S3A: Traces of
 paxilline-sensitive BK currents in freshly isolated cerebral artery smooth
 muscle cells from 12 M-old Col4a1+/+ and Col4a1+/G1344D mice. * S3B: BK
 currents normalized to cell capacitance in freshly isolated cerebral
 artery smooth muscle cells from 12 M-old Col4a1+/+ and Col4a1+/G1344D
 mice. Supplemental_Figure_4_Processed_Data * S4A: mRNA transcript levels
 of genes encoding channels involved in the spontaneous transient outward
 current (Kcnma1, Kcnmb1, Ryr2) and transient inward cation current (Trpm4,
 Itpr1, Itpr2) pathways in isolated cerebral arteries from 12 M-old
 Col4a1+/+ and Col4a1+/G1344D mice measured by ddPCR and presented as fold
 change over control mice. Supplemental_Figure_5_Processed_Data * S5B:
 Representative Ca2+ spark events in cerebral artery smooth muscle cells
 isolated from 4PBA treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and
 untreated Col4a1+/G1344D mice presented as changes in fractional
 fluorescence. * S5C: Ca2+ spark frequency, amplitude, duration (t1/2),
 rise time, and decay (t1/2) in cerebral artery smooth muscle cells
 isolated from 4PBA treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and
 untreated Col4a1+/G1344D mice. * S5E: Frequency of spontaneous transient
 outward currents in freshly isolated cerebral artery smooth muscle cells
 from 4PBA treated 12 M-old Col4a1+/+ and Col4a1+/G1344D and untreated
 Col4a1+/G1344D mice over a range of membrane potentials (-60 to 0 mV). *
 S5F: Amplitude of spontaneous transient outward currents in freshly
 isolated cerebral artery smooth muscle cells from 4PBA treated 12 M-old
 Col4a1+/+ and Col4a1+/G1344D and untreated Col4a1+/G1344D mice over a
 range of membrane potentials (-60 to 0 mV).