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Cannabidiol in Humans—The Quest for Therapeutic Targets

Ambrus Lídia

dahha96
15.03.2019

Content:

  • Ambrus Lídia
  • Human podocytes express functional thermosensitive TRPV channels
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  • View the profiles of people named Lidia Ambrus. Join Facebook to connect with Lidia Ambrus and others you may know. Facebook gives people the power to. Lídia Ambrus of University of Debrecen, Debrecen with expertise in: Physiology, Molecular Biology and Cell Biology. Read 11 publications, and contact Lídia. View Lídia Ambrus' profile on LinkedIn, the world's largest professional community. Lídia has 1 job listed on their profile. See the complete profile on LinkedIn.

    Ambrus Lídia

    Citations Publications citing this paper. Showing of 5 extracted citations. Cellular physiology and biochemistry: Pharmacological inhibition of focal segmental glomerulosclerosis-related, gain of function mutants of TRPC6 channels by semi-synthetic derivatives of larixol.

    Endres , Alexander Staruschenko. References Publications referenced by this paper. Showing of 78 references. New developments in steroid-resistant nephrotic syndrome Moin Saleem.

    Role of altered insulin signaling pathways in the pathogenesis of podocyte malfunction and microalbuminuria. Jauregui , Daniel H. Mintz , Peter H. Mundel , Alessia Fornoni. However, the dose—response relationships were not saturated up to 1. A Representative time courses showing the application of TRPV3 channel activators on differentiated human podocytes in normal buffer.

    Activators and ATP as positive control were applied as shown in the figure. Data are presented as percent of the average of the corresponding control i.

    This normalization was carried out to control the variance of the effectivity between different agonists and make comparable the inhibition evoked by the antagonists in the different groups.

    Response was calculated as the maximal amplitude max. Data are normalized to the average of the untransfected control to indicate the fold change in the expression of TRPV3 protein. However, native TRPV3 and TRPV4 channels were detected at lower molecular weight in podocytes compared to the recombinant channels, which might suggest different degradation of these channels in the two cell lines. These findings are highly consistent with publicly available microarray data of Da Sacco et al.

    Moreover, an additional transcriptome analysis Boerries et al. TRPV1 channels, earlier identified as the capsaicin receptors, are robustly activated by capsaicin in human and rodents.

    Recent research revealed the molecular mechanism of capsaicin binding to TRPV1 channels and identified key amino acid residues, mutations of which decreased capsaicin sensitivity Yang et al. In our case, both mutations in the key residues or the presence of the dominant negative subunit TRPV1b can be a rational explanation for the finding that in spite of the molecular expression of TRPV1 protein, we did not find any functional effect of capsaicin.

    These functional results suggest that, in good accordance with the molecular expression data, TRPV4 channels are the dominantly expressed thermosensitive TRPV channels in human podocytes. The pharmacological identification is uncertain since we lack commercially available, effective and highly specific TRPV3 channel agonists and antagonists. Therefore, although our results suggest that TRPV3 channels are expressed in human podocytes, their function is not clear.

    However, more specific pharmacological tools are needed to validate our results and further dissect the potential role of TRPV3 channels in podocytes. The expression of heat and mechanosensitive TRPV channels has been investigated in the lower urinary tract and in the kidney, ever since their cloning Hayes et al.

    Although there is clear evidence for the role of TRPV1 channels in the control of lower urinary tract functions, its expression in urothelial cells is still under debate see, Franken et al. In the kidney, TRPV4 expression was found overall in the constitutively water impermeable segments of the nephron thin and thick ascending limb, distal convoluted tubule and in the collecting ducts, as well Tian et al.

    In the renal tubular system, an important role in transmitting the effect of tubular flow and osmolarity is attributed to TRPV4 channels Mamenko et al.

    Beyond osmosensation, the TRPV4 channel seems to play an emerging role in the formation of epithelial barrier in several tissues.

    In epidermal keratinocytes of the skin, functionally active TRPV4 channels were found to interact with adherent junction proteins and actin cytoskeleton, enhancing cell—cell junction and tight barrier formation. Moreover, its pharmacological activation augmented tight junction development and barrier recovery Sokabe et al. As podocytes in the outermost layer of the glomeruli form the final barrier to protein loss, their injury is often associated with marked proteinuria syndromes Somlo and Mundel, However, TRPV4 channels can be modulated by intracellular signalling pathways by, among others, angiotensin II signalling Shukla et al.

    All in all, although further studies are needed to clarify the potential role of TRPV channels in podocyte patho physiology, detailed understanding of their pharmacological role in physiological and disease conditions might contribute to the development of future therapies of primary and secondary podocytopathies and related kidney diseases.

    This Declaration acknowledges that this paper adheres to the principles for transparent reporting and scientific rigour of preclinical research recommended by funding agencies, publishers and other organisations engaged with supporting research.

    The antibodies demonstrated high specificity, they detected only the targeted isoform and showed no unspecific staining in the human embryonic kidney originated cell line.

    British Journal of Pharmacology , National Center for Biotechnology Information , U. Journal List Br J Pharmacol v. Published online Nov 2.

    Author information Article notes Copyright and License information Disclaimer. This article has been cited by other articles in PMC. Conclusion and Implications Our results indicate the functional presence of TRPV4 and other thermosensitive TRPV channels in human podocytes and raise the possibility of their involvement in the regulation of glomerular filtration barrier.

    Introduction Among voltage gated ion channels , the transient receptor potential TRP ion channels form a heterogeneous group with diverse functions. Methods Cell cultures The human podocyte cell line provided by Prof.

    Antibodies The following primary antibodies were employed for immunocytochemistry: Western blot Cells were harvested and homogenized in protease inhibitor cocktail 1: Transient overexpression of human recombinant TRPV channels To check the specificity of the antibodies used in Western blot experiments, we transiently overexpressed human recombinant TRPV1—4 proteins in HEKT cells and subjected the cell lysates to Western blotting. Data and statistical analysis The data and statistical analysis comply with the recommendations on experimental design and analysis in pharmacology Curtis et al.

    Nomenclature of targets and ligands Key protein targets and ligands in this article are hyperlinked to corresponding entries in http: Open in a separate window. Conflict of interest The authors declare no conflicts of interest. Declaration of transparency and scientific rigour This Declaration acknowledges that this paper adheres to the principles for transparent reporting and scientific rigour of preclinical research recommended by funding agencies, publishers and other organisations engaged with supporting research.

    Click here for additional data file. Skin Pharmacol Physiol Br J Pharmacol Transient receptor potential vanilloid 4—mediated disruption of the alveolar septal barrier: J Cell Mol Med Molecular fingerprinting of the podocyte reveals novel gene and protein regulatory networks. Acta Physiol Oxf Loss of function of transient receptor potential vanilloid 1 TRPV1 genetic variant is associated with lower risk of active childhood asthma.

    J Biol Chem Functional transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 channels along different segments of the renal vasculature.

    Experimental design and analysis and their reporting: A novel source of cultured podocytes. Cannabinoid actions at TRPV channels: TRPC6 channels and their binding partners in podocytes: Am J Physiol Renal Physiol Transient receptor potential channels in the vasculature.

    Transient receptor potential vanilloid type 1 channels act as mechanoreceptors and cause substance P release and sensory activation in rat kidneys. TRP channels in lower urinary tract dysfunction. J Cell Biochem Molecular determinants of vanilloid sensitivity in TRPV1. Transient receptor potential TRP channels in the airway: Angiotensin II type 1 receptor overexpression in podocytes induces glomerulosclerosis in transgenic rats.

    Eur J Pharmacol TRPV4 channels in the human urogenital tract play a role in cell junction formation and epithelial barrier. Importance of transient receptor potential vanilloid 4 TRPV4 in epidermal barrier function in human skin keratinocytes.

    TRPV1b, a functional human vanilloid receptor splice variant. Deciphering physiological role of the mechanosensitive TRPV4 channel in the distal nephron. The podocyte as a target for therapies — new and old. Nat Rev Nephrol 8: Involvement of TRPV2 activation in intestinal movement through nitric oxide production in mice. Transient receptor potential channels as therapeutic targets. Nat Rev Drug Discov Transient receptor potential channels as drug targets: Cannabidiol exerts sebostatic and antiinflammatory effects on human sebocytes.

    J Clin Invest Human keratinocytes are vanilloid resistant. What do we know about the transient receptor potential vanilloid 2 TRPV2 ion channel?

    Human podocytes express functional thermosensitive TRPV channels

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