title: Histamine type 1-receptor activation by low dose of histamine undermines human glomerular slit diaphragm integrity.
abstract: Histamine has been reported to decrease the ultrafiltration coefficient, which inversely correlates with glomerular permselectivity, however the mechanism(s) underling this effect have never been investigated. This study aimed to assess whether histamine could exert a direct detrimental effect on podocyte permeability and the possible involvement of two key proteins for the glomerular slit diaphragm (SD) integrity, zonula occludens-1 (ZO-1) and P-cadherin. The effect of histamine (100 pM-1000nM) on coloured podocytes junctional integrity was evaluated functionally by a transwell assay of monolayer permeability and morphologically by electron microscopy. Histamine receptor (H1-4R) presence was evaluated at both mRNA (RT-PCR) and protein (immunofluorescence) levels. The Kdand Bmaxvalues for [3H]mepyramine were determined by saturation binding analysis; IP1and cAMP production evoked by histamine were measured by TR-FRET. ZO-1, P-cadherin and vimentin expression was assessed by qRT-PCR and quantitative immunoblotting. Histamine elicited a time- and sigmoidal dose-dependent (maximum effect at 8h, 10nM) increase in podocyte paracellular permeability widening the paracellular spaces. Only H1R was predominantly localised to the podocyte membrane. Consistently, histamine elicited a sigmoidal dose-dependent increase in IP1, but not in cAMP. Histamine exposure evoked a concentration-dependent reduction in both ZO-1 and P-cadherin and a parallel induction of vimentin mRNA expression with a maximum effect after 6h, and protein expression with a maximum effect after 8h. These effects were prevented by the selective H1R antagonist chlorpheniramine. In conclusion, our data demonstrate that histamine, via the H1R, modifies SD morphological and functional integrity, in part, by decreasing the expression of ZO-1 and P-cadherin.
authors.affiliation: Department of Scienza e Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy. Electronic address: eveglia@unito.it.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Veglia
authors.affiliation: Department of Clinical and Experimental Medicine, University of Florence, Viale Pieraccini 6, 50139, Florence, Italy. Electronic address: apini@unito.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pini
authors.affiliation: Department of Scienza e Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy. Electronic address: aldomoggio@gmail.com.
authors.forename: Aldo
authors.initials: A
authors.lastname: Moggio
authors.affiliation: Department of Scienze Mediche, University of Turin, C.So Dogliotti 14, 10126 Turin, Italy. Electronic address: cgrange@unito.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Grange
authors.affiliation: Department of Neuroscience "Rita Levi Montalcini", University of Turin, Via Cherasco 15, 10126 Turin, Italy. Electronic address: fpremoselli@unito.it.
authors.forename: Federica
authors.initials: F
authors.lastname: Premoselli
authors.affiliation: Department of Scienza e Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy. Electronic address: acrosa@unito.it.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Miglio
authors.affiliation: Department of Pharmacology, Medical School, University of Athens, M. Asias 75, GR-115 27 Athens, Greece. Electronic address: aityliga@med.uoa.gr.
authors.forename: Katerina
authors.initials: K
authors.lastname: Tiligada
authors.affiliation: Department of Scienza e Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy. Electronic address: rfantozzi@unito.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Fantozzi
authors.affiliation: School of Biological and Biomedical Science, Durham University, Durham DH13LE, UK. Electronic address: paul.chazot@durham.ac.uk.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Chazot
authors.affiliation: Department of Scienza e Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy; School of Biological and Biomedical Science, Durham University, Durham DH13LE, UK.
authors.forename: Arianna Carolina
authors.initials: AC
authors.lastname: Rosa
mesh_terms: D015820:Cadherins / Q000032:analysis / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D006632:Histamine / Q000009:adverse effects*; D017442:Histamine Agonists / Q000009:adverse effects*; D006801:Humans; D007678:Kidney Glomerulus / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D050199:Podocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D011969:Receptors, Histamine H1 / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein / Q000032:analysis / Q000378:metabolism
chemical_list: D015820:Cadherins; D017442:Histamine Agonists; D011969:Receptors, Histamine H1; D062826:Zonula Occludens-1 Protein; D006632:Histamine
keywords: Chlorpheniramine maleate PubChem CID 5281068; Diphenhydramine Pubmed CID 3100; Histamine; Histamine dihydrochloride PubChem CID 5818; Histamine receptors; Junction integrity; Paracellular permeability; Podocytes; [(3)H]mepyramine PubChem CID 656400