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Protein Kinase C Family Members as a Target for Regulation of Blood–Brain Barrier Na,K,2Cl-Cotransporter During In Vitro Stroke Conditions and Nicotine Exposure

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Purpose

The aim of the study is to identify specific protein kinase C (PKC) isoforms involvement in K+ transport mediated at altered blood–brain barrier (BBB) response to stroke conditions with prior nicotine exposure, which provides ways to intervene pharmacologically in PKC-mediated molecular pathways that could lead to effective treatment for smoking stroke patients.

Methods

Changes in PKC isoform levels were studied in the cytosolic and membrane fractions of bovine brain microvessel endothelial cells subjected to stroke conditions as well as nicotine/cotinine exposure. Furthermore, abluminal Na,K,2Cl-cotransporter (NKCC) activity regulated by specific conventional PKC isoform activators and inhibitors was investigated using rubidium (86Rb) uptake studies.

Results

Membrane-bound PKCα, PKCβI, and PKCɛ levels were increased after 6 h hypoxia/aglycemia, and this was attenuated by 24-h nicotine/cotinine exposure. Interestingly, membrane-bound PKCγ protein level was decreased after 6 h hypoxia/aglycemia and increased by 24-h nicotine/cotinine exposure. 86Rb uptake studies showed that basolateral NKCC activity was down-regulated by both a conventional PKC inhibitor and specific inhibitors for PKCα, PKCβ, and PKCɛ and was up-regulated by an activator of conventional PKCs during 6-h hypoxia/aglycemia treatment.

Conclusion

Specific PKC inhibitors or activators might be designed to individualize stroke therapies and improve health outcome for smokers by rebalancing ion transport into and out of the brain.

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References

  1. J. S. Gill M. J. Shipley S. A. Tsementzis R. Hornby S. K. Gill E. R. Hitchcock D. G. Beevers (1989) ArticleTitleCigarette smoking. A risk factor for hemorrhagic and nonhemorrhagic stroke Arch. Intern. Med. 149 2053–2057 Occurrence Handle10.1001/archinte.149.9.2053 Occurrence Handle1:STN:280:BiaA2szjtlM%3D Occurrence Handle2774782

    Article  CAS  PubMed  Google Scholar 

  2. L. Wang M. Kittaka N. Sun S. Schreiber B. V. Zlokovic (1997) ArticleTitleChronic nicotine treatment enhances focal ischemic brain injury and depletes free pool of brain microvascular tissue plasminogen activator in rats J. Cereb. Blood Flow Metab. 17 136–147 Occurrence Handle1:CAS:528:DyaK2sXhvV2jt7o%3D Occurrence Handle9040492

    CAS  PubMed  Google Scholar 

  3. T. A. Brock C. Brugnara M. Canessa M. A. Gimbrone (1986) ArticleTitleBradykinin and vasopressin stimulate Na,K-2Cl-cotransport in cultured endothelial cells Am. J. Physiol., Cell Physiol. 250 C888–C895 Occurrence Handle1:CAS:528:DyaL28XksFGhtbg%3D

    CAS  Google Scholar 

  4. T. J. Abbruscato S. P. Lopez K. S. Mark B. T. Hawkins T. P. Davis (2002) ArticleTitleNicotine and cotinine modulate cerebral microvascular permeability and protein expression of ZO-1 through nicotinic acetylcholine receptors expressed on brain endothelial cells J. Pharm. Sci. 91 2525–2538 Occurrence Handle10.1002/jps.10256 Occurrence Handle1:CAS:528:DC%2BD38XptVeksLY%3D Occurrence Handle12434396

    Article  CAS  PubMed  Google Scholar 

  5. T. J. Abbruscato S. P. Lopez K. Roder J. R. Paulson (2004) ArticleTitleRegulation of blood–brain barrier Na,K,2Cl-cotransporter through phosphorylation during in vitro stroke conditions andnicotine exposure J. Pharmacol. Exp. Ther. 310 459–468 Occurrence Handle10.1124/jpet.104.066274 Occurrence Handle1:CAS:528:DC%2BD2cXmtVyks78%3D Occurrence Handle15051802

    Article  CAS  PubMed  Google Scholar 

  6. M. E. O'Donnell A. Martinez D. Sun (1995) ArticleTitleCerebral microvascular endothelial cell Na–K–Cl contransport: regulation by astrocyte-conditioned medium Am. J. Physiol. 268 C747–C754 Occurrence Handle7900779

    PubMed  Google Scholar 

  7. D. Sun M. E. O'Donnell (1996) ArticleTitleAstroglial-mediated phosphorylation of the Na–K–Cl contransporter in brain microvessel endothelial cells Am. J. Physiol. 271 C620–C627 Occurrence Handle1:CAS:528:DyaK28XlsV2ksrg%3D Occurrence Handle8770003

    CAS  PubMed  Google Scholar 

  8. P. Vigne A. L. Farre C. Frelin (1994) ArticleTitleNa+–K+–Cl− cotransporter of brain capillary endothelial cells J. Biol. Chem. 269 19925–19930 Occurrence Handle1:CAS:528:DyaK2cXltlSktrk%3D Occurrence Handle8051075

    CAS  PubMed  Google Scholar 

  9. N. Kawai R. M. McCarron M. Spatz (1996) ArticleTitleEffect of hypoxia on Na+–K+–Cl-cotransport in cultured brain capillary endothelial cells of the rat J. Neurochem. 66 2572–2579 Occurrence Handle1:CAS:528:DyaK28XjtFCrsr8%3D Occurrence Handle8632184

    CAS  PubMed  Google Scholar 

  10. T. Wieloch M. Cardell H. Bingren J. Zivin T. Saitoh (1991) ArticleTitleChanges in the activity of protein kinase C and the differential subcellular redistribution of its isozymes in the rat striatum during and following transient forebrain ischemia J. Neurochem. 56 1227–1235 Occurrence Handle1:CAS:528:DyaK3MXitVCqurw%3D Occurrence Handle2002338

    CAS  PubMed  Google Scholar 

  11. J. Krupinski M. A. Slevin P. Kumar J. Gaffney J. Kaluza (1998) ArticleTitleProtein kinase C expression and activity in the human brain after ischemic stroke Acta Neurobiol. Exp. 58 13–21 Occurrence Handle1:STN:280:DyaK1c3ksVSiug%3D%3D

    CAS  Google Scholar 

  12. F. L. Huang Y. Yoshida H. Nakabayashi W. S. Young K. P. Huang (1988) ArticleTitleImmunocytochemical localization of protein kinase C isozymes in rat brain J. Neurosci. 8 4734–4744 Occurrence Handle1:CAS:528:DyaL1MXlsVygsQ%3D%3D Occurrence Handle2462028

    CAS  PubMed  Google Scholar 

  13. M. S. Shearman Z. Naor K. Sekiguchi A. Kishimoto Y. Nishizuka (1989) ArticleTitleSelective activation of the g-subspecies of protein kinase C from bovine cerebellum by arachidonic acid and its lipoxygenase metabolites FEBS Lett. 243 177–182 Occurrence Handle10.1016/0014-5793(89)80125-5 Occurrence Handle1:CAS:528:DyaL1MXhs12juro%3D Occurrence Handle2492951

    Article  CAS  PubMed  Google Scholar 

  14. T. R. Yerby T. Vibat D. Sun J. A. Payne M. E. O'Donnell (1997) ArticleTitleMolecular characterization of the Na,K,2Cl-cotransporter of bovine aortic endothelial cells Am. J. Physiol. 273 C188–C197 Occurrence Handle1:CAS:528:DyaK2sXlt1Sku7Y%3D Occurrence Handle9252456

    CAS  PubMed  Google Scholar 

  15. K. L. Audus R. T. Borchardt (1987) ArticleTitleBovine brain microvessel endothelial cell monolayers as a model system for the blood–brain barrier Ann. N.Y. Acad. Sci. 507 9–18 Occurrence Handle1:STN:280:BieC287pslQ%3D Occurrence Handle3442370

    CAS  PubMed  Google Scholar 

  16. R. R. Bosch M. Bazuine M. M. Wake P. N. Span A. J. Olthaar A. Schurmann J. A. Maassen A. R. Hermus P. H. Willems C. G. Sweep (2003) ArticleTitleInhibition of protein kinase CbetaII increases glucose uptake in 3T3-L1 adipocytes through elevated expression of glucose transporter 1 at the plasma membrane Mol. Endocrinol. 17 1230–1239 Occurrence Handle10.1210/me.2003-0057 Occurrence Handle1:CAS:528:DC%2BD3sXlt1Krur0%3D Occurrence Handle12702734

    Article  CAS  PubMed  Google Scholar 

  17. N. E. Owen M. L. Prastein (1985) ArticleTitleNa/K/Cl contransport in cultured human fibroblasts J. Biol. Chem. 260 1445–1451 Occurrence Handle1:CAS:528:DyaL2MXptlGrsw%3D%3D Occurrence Handle2981857

    CAS  PubMed  Google Scholar 

  18. D. H. Hryciw C. A. Pollock P. Poronnik (2005) ArticleTitlePKC-alpha-mediated remodeling of the actin cytoskeleton is involved in constitutive albumin uptake by proximal tubule cells Am. J. Physiol., Renal Physiol. 288 F1227–F1235 Occurrence Handle10.1152/ajprenal.00428.2003 Occurrence Handle1:CAS:528:DC%2BD2MXlsVymt70%3D

    Article  CAS  Google Scholar 

  19. S. Y. Yuan E. E. Ustinova M. H. Wu J. H. Tinsley W. Xu F. L. Korompai A. C. Taulman (2000) ArticleTitleProtein kinase C activation contributes to microvascular barrier dysfunction in the heart at early stages of diabetes Circ. Res. 87 412–417 Occurrence Handle1:CAS:528:DC%2BD3cXms1ymsrw%3D Occurrence Handle10969040

    CAS  PubMed  Google Scholar 

  20. N. Shiroshita M. Musashi K. Sakurada K. Kimura Y. Tsuda S. Ota H. Iwasaki T. Miyazaki T. Kato H. Miyazaki A. Shimosaka M. Asaka (2001) ArticleTitleInvolvement of protein kinase C-epsilon in signal transduction of thrombopoietin in enhancement of interleukin-3-dependent proliferation of primitive hematopoietic progenitors J. Pharmacol. Exp. Ther. 297 868–875 Occurrence Handle1:CAS:528:DC%2BD3MXjvVSiuro%3D Occurrence Handle11356906

    CAS  PubMed  Google Scholar 

  21. J. A. Johnson M. O. Gray C. H. Chen D. Mochly-Rosen (1996) ArticleTitleA protein kinase C translocation inhibitor as an isozyme-selective antagonist of cardiac function J. Biol. Chem. 271 24962–24966 Occurrence Handle1:CAS:528:DyaK28XmtFens74%3D Occurrence Handle8798776

    CAS  PubMed  Google Scholar 

  22. M. Yedovitzky D. Mochly-Rosen J. A. Johnson M. O. Gray D. Ron E. Abramovitch E. Cerasi R. Nesher (1997) ArticleTitleTranslocation inhibitors define specificity of protein kinase C isoenzymes in pancreatic beta-cells J. Biol. Chem. 272 1417–1420 Occurrence Handle1:CAS:528:DyaK2sXls1ektQ%3D%3D Occurrence Handle8999804

    CAS  PubMed  Google Scholar 

  23. K. C. Das X. L. Guo C. W. White (1998) ArticleTitleProtein kinase Cdelta-dependent induction of manganese superoxide dismutase gene expression by microtubule-active anticancer drugs J. Biol. Chem. 273 34639–34645 Occurrence Handle10.1074/jbc.273.51.34639 Occurrence Handle1:CAS:528:DyaK1MXntlGk Occurrence Handle9852137

    Article  CAS  PubMed  Google Scholar 

  24. T. J. Abbruscato T. P. Davis (1999) ArticleTitleCombination of hypoxia/aglycemia compromises in vitro BBB integrity J. Pharmacol. Exp. Ther. 289 668–675 Occurrence Handle1:CAS:528:DyaK1MXislGjtbY%3D Occurrence Handle10215638

    CAS  PubMed  Google Scholar 

  25. T. J. Abbruscato T. P. Davis (1999) ArticleTitleProtein expression of brain endothelial cell E-cadherin after hypoxia/aglycemia: influence of astrocyte contact Brain Res. 842 277–286 Occurrence Handle10.1016/S0006-8993(99)01778-3 Occurrence Handle1:CAS:528:DyaK1MXmsVWgu7g%3D Occurrence Handle10526124

    Article  CAS  PubMed  Google Scholar 

  26. S. Umar J. H. Sellin A. P. Morris (2000) ArticleTitleIncreased nuclear translocation of catalytically active PKC-zeta during mouse colonocyte hyperproliferation Am. J. Physiol.: Gasterointest. Liver Physiol. 279 G223–G237 Occurrence Handle1:CAS:528:DC%2BD3cXltl2jt7w%3D

    CAS  Google Scholar 

  27. J. M. Russell (2000) ArticleTitleSodium–potassium–chloride cotransport Physiol. Rev. 80 211–276 Occurrence Handle1:CAS:528:DC%2BD3cXmtl2jug%3D%3D Occurrence Handle10617769

    CAS  PubMed  Google Scholar 

  28. M. E. O'Donnell A. Martinez D. Sun (1995) ArticleTitleEndothelial Na–K–Cl cotransport regulation by tonicity and hormones: phosphorylation of cotransport protein Am. J. Physiol., Cell Physiol. 269 C1513–C1523

    Google Scholar 

  29. B. J. Padanilam (2001) ArticleTitleInduction and subcellular localization of protein kinase C isozymes following renal ischemia Kidney Int. 59 1789–1797 Occurrence Handle10.1046/j.1523-1755.2001.0590051789.x Occurrence Handle1:CAS:528:DC%2BD3MXjs1ehsbs%3D Occurrence Handle11318949

    Article  CAS  PubMed  Google Scholar 

  30. H. Mellor P. J. Parker (1998) ArticleTitleThe extended protein kinase C superfamily Biochem. J. 332 281–292 Occurrence Handle1:CAS:528:DyaK1cXkt1Sqs74%3D Occurrence Handle9601053

    CAS  PubMed  Google Scholar 

  31. M. S. Erclik J. Mitchell (2002) ArticleTitleThe role of protein kinase C-δ in PTH stimulation of IGF-binding protein-5 mRNA in UMR-106-01 cells Am. J. Physiol.: Endocrinol. Metab. 282 E534–E541 Occurrence Handle1:CAS:528:DC%2BD38Xit1GiurY%3D

    CAS  Google Scholar 

  32. M. Csukai D. Mochly-Rosen (1999) ArticleTitlePharmacologic modulation of protein kinase C isozymes: the role of RACKs and subcellular localization Pharmacol. Res. 39 253–259 Occurrence Handle10.1006/phrs.1998.0418 Occurrence Handle1:CAS:528:DyaK1MXisFGjtLw%3D Occurrence Handle10208754

    Article  CAS  PubMed  Google Scholar 

  33. C. Dempsey A. C. Newton D. Mochly-Rosen A. P. Fields M. E. Reyland P. A. Nissel R. O. Messing (2000) ArticleTitleProtein kinase C isozymes and the regulation of diverse cell responses Am. J. Physiol., Lung Cell. Mol. Physiol. 279 L429–L438 Occurrence Handle1:CAS:528:DC%2BD3cXmvVGrtLo%3D

    CAS  Google Scholar 

  34. E. A. Brownson T. J. Abbruscato T. J. Gillespie V. J. Hruby T. P. Davis (1994) ArticleTitleEffect of peptidases at the blood–brain barrier on the permeability of enkephalin J. Pharmacol. Exp. Ther. 270 675–680 Occurrence Handle1:CAS:528:DyaK2cXlslSqsbw%3D Occurrence Handle7915319

    CAS  PubMed  Google Scholar 

  35. K. Domanska-Janik T. Zalewska (1992) ArticleTitleEffect of brain ischemia on protein kinase C J. Neurochem. 58 1432–1439 Occurrence Handle1:CAS:528:DyaK38XhvFajsb4%3D Occurrence Handle1548477

    CAS  PubMed  Google Scholar 

  36. M. Cardell T. Wieloch (1993) ArticleTitleTime course of the translocation and inhibition of protein kinase C during complete cerebral ischemia in the rat J. Neurochem. 61 1308–1314 Occurrence Handle1:CAS:528:DyaK3sXms1yjsb8%3D Occurrence Handle8376989

    CAS  PubMed  Google Scholar 

  37. R. Selvatici S. Marino C. Piubello D. Rodi L. Beani E. Gandini A. Siniscalchi (2002) ArticleTitleProtein kinase C activity, translocation, and selective isoform subcellular redistribution in the rat cerebral cortex after in vitro ischemia J. Neurosci. Res. 71 64–71

    Google Scholar 

  38. M. E. O'Donnell L. Tran T. I. Lam X. B. Liu S. E. Anderson (2004) ArticleTitleBumetanide inhibition of the blood–brain barrier Na–K–Cl cotransporter reduces edema formation in the rat middle cerebral artery occlusion model of stroke J. Cereb. Blood Flow Metab. 24 1046–1056 Occurrence Handle15356425

    PubMed  Google Scholar 

  39. Y. Yan R. J. Dempsey A. Flemmer B. Forbush D. Sun (2003) ArticleTitleInhibition of Na–K–Cl cotransporter during focal cerebral ischemia decreases edema and neuronal damage Brain Res. 961 22–31 Occurrence Handle10.1016/S0006-8993(02)03832-5 Occurrence Handle1:CAS:528:DC%2BD3sXivVGjsw%3D%3D Occurrence Handle12535773

    Article  CAS  PubMed  Google Scholar 

  40. H. Chen J. Luo D. B. Kintner G. E. Shull D. Sun (2005) ArticleTitleNa+-dependent chloride transporter (NKCC1)-null mice exhibit less gray and white matter damage after focal cerebral ischemia J. Cereb. Blood Flow Metab. 25 54–66 Occurrence Handle1:CAS:528:DC%2BD2MXht1KksL8%3D Occurrence Handle15678112

    CAS  PubMed  Google Scholar 

  41. J. R. Paulson K. E. Roder G. McAfee D. D. Allen C. J. Schyf ParticleVan der T. J. Abbruscato (2006) ArticleTitleTobacco smoke chemicals attenuate brain-to-blood potassium transport mediated by the Na,K,2Cl-cotransporter during hypoxia-reoxygenation J. Pharmacol. Exp. Ther. 316 248–254 Occurrence Handle1:CAS:528:DC%2BD28XhslCjsg%3D%3D Occurrence Handle16174793

    CAS  PubMed  Google Scholar 

  42. N. L. Benowitz (1997) ArticleTitleThe role of nicotine in smoking-related cardiovascular disease Prev. Med. 26 412–417 Occurrence Handle10.1006/pmed.1997.0175 Occurrence Handle1:CAS:528:DyaK2sXltlKluro%3D Occurrence Handle9245658

    Article  CAS  PubMed  Google Scholar 

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Acknowledgment

This work was supported by a National Institution of Health grant NS046526.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter Drive, Amarillo, Texas, 79106, USA

    Tianzhi Yang, Karen E. Roder, G. Jayarama Bhat, Thomas J. Thekkumkara & Thomas J. Abbruscato

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  1. Tianzhi Yang
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  2. Karen E. Roder
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  3. G. Jayarama Bhat
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Correspondence to Thomas J. Abbruscato.

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Yang, T., Roder, K.E., Bhat, G.J. et al. Protein Kinase C Family Members as a Target for Regulation of Blood–Brain Barrier Na,K,2Cl-Cotransporter During In Vitro Stroke Conditions and Nicotine Exposure. Pharm Res 23, 291–302 (2006). https://doi.org/10.1007/s11095-005-9143-2

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