Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome
Mounting evidence supports the role of the endocannabinoid system in neurophysiology, including blood–brain barrier (BBB) function. Recent work has demonstrated that activation of endocannabinoid receptors can mitigate insults to the BBB during neurological disorders like traumatic brain injury, cor...
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| Format: | Article |
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MDPI AG
2023-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/1/531 |
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| author | Aidan A. Levine Erika Liktor-Busa Shreya Balasubramanian Seph M. Palomino Anya M. Burtman Sarah A. Couture Austin A. Lipinski Paul R. Langlais Tally M. Largent-Milnes |
| author_facet | Aidan A. Levine Erika Liktor-Busa Shreya Balasubramanian Seph M. Palomino Anya M. Burtman Sarah A. Couture Austin A. Lipinski Paul R. Langlais Tally M. Largent-Milnes |
| author_sort | Aidan A. Levine |
| collection | DOAJ |
| description | Mounting evidence supports the role of the endocannabinoid system in neurophysiology, including blood–brain barrier (BBB) function. Recent work has demonstrated that activation of endocannabinoid receptors can mitigate insults to the BBB during neurological disorders like traumatic brain injury, cortical spreading depression, and stroke. As alterations to the BBB are associated with worsening clinical outcomes in these conditions, studies herein sought to examine the impact of endocannabinoid depletion on BBB integrity. Barrier integrity was investigated in vitro via bEnd.3 cell monolayers to assess endocannabinoid synthesis, barrier function, calcium influx, junctional protein expression, and proteome-wide changes. Inhibition of 2-AG synthesis using DAGLα inhibition and siRNA inhibition of DAGLα led to loss of barrier integrity via altered expression of VE-cadherin, which could be partially rescued by exogenous application of 2-AG. Moreover, the deleterious effects of DAGLα inhibition on BBB integrity showed both calcium and PKC (protein kinase C)-dependency. These data indicate that disruption of 2-AG homeostasis in brain endothelial cells, in the absence of insult, is sufficient to disrupt BBB integrity thus supporting the role of the endocannabinoid system in neurovascular disorders. |
| first_indexed | 2024-03-08T15:04:42Z |
| format | Article |
| id | doaj.art-8791bae7b6cc4b64bcc65157dedb1cf1 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T15:04:42Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8791bae7b6cc4b64bcc65157dedb1cf12024-01-10T14:59:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-12-0125153110.3390/ijms25010531Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-ProteomeAidan A. Levine0Erika Liktor-Busa1Shreya Balasubramanian2Seph M. Palomino3Anya M. Burtman4Sarah A. Couture5Austin A. Lipinski6Paul R. Langlais7Tally M. Largent-Milnes8Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USADivision of Endocrinology, Department of Medicine, College of Medicine, University of Arizona, Tucson, AZ 85724, USADivision of Endocrinology, Department of Medicine, College of Medicine, University of Arizona, Tucson, AZ 85724, USADepartment of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ 85724, USAMounting evidence supports the role of the endocannabinoid system in neurophysiology, including blood–brain barrier (BBB) function. Recent work has demonstrated that activation of endocannabinoid receptors can mitigate insults to the BBB during neurological disorders like traumatic brain injury, cortical spreading depression, and stroke. As alterations to the BBB are associated with worsening clinical outcomes in these conditions, studies herein sought to examine the impact of endocannabinoid depletion on BBB integrity. Barrier integrity was investigated in vitro via bEnd.3 cell monolayers to assess endocannabinoid synthesis, barrier function, calcium influx, junctional protein expression, and proteome-wide changes. Inhibition of 2-AG synthesis using DAGLα inhibition and siRNA inhibition of DAGLα led to loss of barrier integrity via altered expression of VE-cadherin, which could be partially rescued by exogenous application of 2-AG. Moreover, the deleterious effects of DAGLα inhibition on BBB integrity showed both calcium and PKC (protein kinase C)-dependency. These data indicate that disruption of 2-AG homeostasis in brain endothelial cells, in the absence of insult, is sufficient to disrupt BBB integrity thus supporting the role of the endocannabinoid system in neurovascular disorders.https://www.mdpi.com/1422-0067/25/1/531endocannabinoidblood–brain barrier2-AGDAGLαVE-cadherin |
| spellingShingle | Aidan A. Levine Erika Liktor-Busa Shreya Balasubramanian Seph M. Palomino Anya M. Burtman Sarah A. Couture Austin A. Lipinski Paul R. Langlais Tally M. Largent-Milnes Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome International Journal of Molecular Sciences endocannabinoid blood–brain barrier 2-AG DAGLα VE-cadherin |
| title | Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome |
| title_full | Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome |
| title_fullStr | Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome |
| title_full_unstemmed | Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome |
| title_short | Depletion of Endothelial-Derived 2-AG Reduces Blood-Endothelial Barrier Integrity via Alteration of VE-Cadherin and the Phospho-Proteome |
| title_sort | depletion of endothelial derived 2 ag reduces blood endothelial barrier integrity via alteration of ve cadherin and the phospho proteome |
| topic | endocannabinoid blood–brain barrier 2-AG DAGLα VE-cadherin |
| url | https://www.mdpi.com/1422-0067/25/1/531 |
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