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Journal of Molecular Medicine

, Volume 98, Issue 1, pp 97–110 | Cite as

Selective protection of murine cerebral Gi/o-proteins from inactivation by parenterally injected pertussis toxin

  • Salvador Castaneda Vega
  • Veronika Leiss
  • Roland Piekorz
  • Carsten Calaminus
  • Katja Pexa
  • Marta Vuozzo
  • Andreas M. Schmid
  • Vasudharani Devanathan
  • Christian Kesenheimer
  • Bernd J. Pichler
  • Sandra Beer-Hammer
  • Bernd NürnbergEmail author
Original Article

Abstract

Pertussis toxin (PTX) is a potent virulence factor in patients suffering from whooping cough, but in its detoxified version, it is applied for vaccination. It is thought to contribute to the pathology of the disease including various CNS malfunctions. Based on its enzymatic activity, PTX disrupts GPCR-dependent signaling by modifying the α-subunit of heterotrimeric Gi/o-proteins. It is also extensively used as a research tool to study neuronal functions in vivo and in vitro. However, data demonstrating the penetration of PTX from the blood into the brain are missing. Here, we examined the Gαi/o-modifying activity of PTX in murine brains after its parenteral application. Ex vivo biodistribution analysis of [124I]-PTX displayed poor distribution to the brain while relatively high concentrations were visible in the pancreas. PTX affected CNS and endocrine functions of the pancreas as shown by open-field and glucose tolerance tests, respectively. However, while pancreatic islet Gαi/o-proteins were modified, their neuronal counterparts in brain tissue were resistant towards PTX as indicated by different autoradiographic and immunoblot SDS-PAGE analyses. In contrast, PTX easily modified brain Gαi/o-proteins ex vivo. An attempt to increase BBB permeability by application of hypertonic mannitol did not show PTX activity on neuronal G proteins. Consistent with these findings, in vivo MRI analysis did not point to an increased blood-brain barrier (BBB) permeability following PTX treatment. Our data demonstrate that the CNS is protected from PTX. Thus, we hypothesize that the BBB hinders PTX to penetrate into the CNS and to deliver its enzymatic activity to brain Gαi/o-proteins.

Key messages

  • i.p. applied PTX is poorly retained in the brain while reaches high concentration in the pancreas.

  • Pancreatic islet Gαi/o- but not cerebral Gαi/o-proteins are modified by i.p. administered PTX.

  • i/o-proteins from isolated cerebral cell membranes were easily modified by PTX ex vivo.

  • CNS is protected from i.p. administered PTX.

  • PTX does not permeabilize the BBB.

Keywords

Pertussis toxin/PTX G-proteins Biodistribution CNS Blood-brain barrier 

Abbreviations

BBB

Blood-brain barrier

b.w.

Body weight

CNS

Central nervous system

DDT

Dithiothreitol

GEF

Guanine nucleotide exchange factor

G proteins

Guanine nucleotide–binding proteins

GPCR

G protein–coupled receptors

ID

Injected dose

i.p.

Intraperitoneal

MRI

Magnetic resonance imaging

PMSF

Phenylmethanesulfonyl fluoride

PTX

Pertussis toxin

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

WBC

White blood cells

wt

Wild type

Notes

Acknowledgments

The authors would like to thank the following people for their assistance in this research: Maren Harant, Daniel Bukala, Renate Riehle, and Sandra Schwegmann.

Funding information

This work was supported by grants of the Deutsche Forschungsgemeinschaft (DFG; NU 53/9-2 to B.N. and V.L. and NU 53/13-1 to B.N.), Deutscher Akademischer Austauschdienst (DAAD), Else Kröner-Fresenius, and the Werner Siemens Foundation.

Compliance with ethical standards

Animal studies were approved by the local ethics committees (Regierungspräsidium Düsseldorf (50.05-230-45/04, 8.87-50.10.34.08.018) and Tübingen (PH10/13, PH1/11, PH4/19), which follow the 2010/63/EU Directive.

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Salvador Castaneda Vega
    • 1
    • 2
  • Veronika Leiss
    • 3
  • Roland Piekorz
    • 4
  • Carsten Calaminus
    • 1
  • Katja Pexa
    • 4
  • Marta Vuozzo
    • 1
  • Andreas M. Schmid
    • 1
  • Vasudharani Devanathan
    • 3
    • 5
  • Christian Kesenheimer
    • 1
  • Bernd J. Pichler
    • 1
    • 2
  • Sandra Beer-Hammer
    • 3
  • Bernd Nürnberg
    • 3
    • 6
    Email author
  1. 1.Werner Siemens Imaging Center, Department of Preclinical Imaging and RadiopharmacyEberhard Karls University Tübingen and University Medical CenterTübingenGermany
  2. 2.Department of Nuclear Medicine and Clinical Molecular ImagingEberhard Karls UniversityTübingenGermany
  3. 3.Department of Pharmacology and Experimental Therapy, Institute for Experimental and Clinical Pharmacology and Toxicology, Interfaculty Center for Pharmacogenomics and Drug ResearchEberhard Karls University TübingenTübingenGermany
  4. 4.Institute for Biochemistry and Molecular Biology II, Medical FacultyHeinrich Heine University DüsseldorfDüsseldorfGermany
  5. 5.Neuroscience Lab, Department of BiologyIndian Institute of Science Education and Research (IISER)TirupatiIndia
  6. 6.Department of Toxicology, Institute for Experimental and Clinical Pharmacology and ToxicologyEberhard Karls University Tübingen, and University Medical CenterTübingenGermany

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