Impaired intracellular Ca2+ signaling contributes to age-related cerebral small vessel disease in Col4a1 mutant mice
Data files
Jan 11, 2024 version files 18.09 MB
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Figure_1_Processed_Data.xlsx
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Figure_2_Processed_Data.xlsx
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Figure_3_Processed_Data.xlsx
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Figure_4_Processed_Data.xlsx
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Figure_5_Processed_Data.xlsx
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README.md
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Supplemental_Figure_1_Processed_Data.xlsx
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Supplemental_Figure_2_Processed_Data.xlsx
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Supplemental_Figure_3_Processed_Data.xlsx
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Supplemental_Figure_4_Processed_Data.xlsx
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Supplemental_Figure_5_Processed_Data.xlsx
Abstract
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.
README: 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).