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Immunization with DISC1 protein in an animal model of ADHD influences behavior and excitatory amino acids in prefrontal cortex and striatum

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Abstract

The Disrupted-in-schizophrenia 1 (DISC1) gene is involved in vulnerability to neuropsychiatric disorders. Naples high-excitability (NHE) rat model neuropsychiatric problems characterized by an unbalanced mesocortical dopamine system. Here, we assessed behavioral and neurochemical effects of immunization against multimeric rat DISC1 protein in adult NHE rats, an animal model of attention-deficit hyperactivity disorder and their Random-Bred (NRB) controls. Males of both lines received subcutaneous injections of vehicle (PB), adjuvant only (AD) or recombinant rat DISC1 protein purified from E. coli, suspended in AD (anti-DISC1) at age of 30, 45 and 60 postnatal days (pnd). At 75 pnd, the rats were exposed to a Làt maze and 2 days later to an Olton eight-arm radial maze, and horizontal (HA) and vertical activities (VA) were monitored. Non-selective (NSA) and selective spatial attention (SSA) were monitored in the Làt and in the Olton maze by duration of rearings and working memory, respectively. Post mortem neurochemistry in the prefrontal cortex (PFc), dorsal (DS) and ventral (VS) striatum of l-Glutamate, l-Aspartate and l-Leucine was performed. All immunized rats showed a clear humoral IgM (but not IgG) immune response against the immunogen, indicating that immunological self-tolerance to DISC1 can be overcome by immunization. NHE rats exhibited a higher unspecific IgM response to adjuvant, indicating an immunological abnormality. The sole anti-DISC1 immunization-specific behavioral in the NHE rats was an increased horizontal activity in the Làt maze. Adjuvant treatment increased vertical activity in both lines, but in the NRB controls it increased rearing and decreased horizontal activity. Liquid chromatography/tandem mass spectrometry analysis of soluble or membrane-trapped neurotransmitters aspartate, glutamate and leucine revealed increased soluble aspartate levels in the ventral striatum of NRB controls after anti-DISC1 immunization. Immune activation by adjuvant independent of simultaneous DISC1 immunization led to other specific changes in NHE and control NRB rats. In DISC1-immunized NHE rats, horizontal activity in Lat maze correlated with membrane-trapped glutamate in PFc and in the NRB rats, duration of rearing in Olton maze correlated with membrane-trapped glutamate in PFc and aspartate in dorsal striatum. In addition to non-specific immune activation (by AD), the postnatal anti-DISC1 immune treatment led to behavioral changes related to mechanisms of activity and attention and had influenced amino acids and synaptic markers in striatum and neocortex in the adult NHE as well as control animals.

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Abbreviations

AD:

Adjuvant

ADHD:

Attention-deficit hyperactivity disorder

anti-DISC1:

Anti-DISC1 protein immunization

CE:

Collision energy

DA:

Dopamine

DAT:

Dopamine transporter

DISC1:

Disrupted-in-schizophrenia 1

DS:

Dorsal striatum

EAA:

Excitatory amino acids

FE:

Position of first repetitive arm visit

HA:

Horizontal activity

MTF:

Membrane-trapped form

NHE:

Naples high-excitability rat

NRB:

Naples random bred

NSA:

Non-selective attention

NVTC:

Number of arm visits before all arms were visited

PB:

Phosphate buffer vehicle

PFc:

Prefrontal cortex

Pnd:

Post natal days

RT:

Retention time

Sc:

Subcutaneous

SF:

Soluble form

SRM:

Selected reaction monitoring

SSA:

Selective spatial attention

VA:

Vertical activity

VS:

Ventral striatum

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Acknowledgments

This research was supported by the Young Investigator Project 2009–2012 from the Italian Ministry of Health to LAR and funding support by the Brain Behavior and Research Foundation (NARSAD Independent Investigator Award #20350), NEURON-ERANET (“DISCover”, BMBF 01EW1003), and EU-FP MC-ITN “IN-SENS” #607616) to C. K. M. A. de Souza Silva was supported by a Heisenberg Fellowship SO 1032/5-1 and EU-FP7 (MC-ITN-“In-SENS”—# 607616). LAR, G. Boatto and A. Tino share equal first author dignity. C. Korth and AGS share last author dignity.

Conflict of interest

None of the authors declare any conflict of interest.

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Authors and Affiliations

  1. Department of Experimental Medicine, School of Medicine, Second University of Naples, S. Andrea delle Dame, 7, 80138, Naples, Italy

    L. A. Ruocco, C. Treno, U. A. Gironi Carnevale & A. G. Sadile

  2. Animal facility, I. N. T. G. Pascale, Naples, Italy

    C. Arra

  3. Department of Chemistry and Pharmacy, Sassari University, Sassari, Italy

    G. Boatto & M. Nieddu

  4. Istituto di Cibernetica “Eduardo Caianiello” ICIB CNR, Pozzuoli, Italy

    C. Pagano & A. Tino

  5. Department Neuropathology, University of Duesseldorf, Medical School, Duesseldorf, Germany

    M. Michel, I. Prikulis & C. Korth

  6. Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy

    E. Carboni

  7. Center for Behavioral Neuroscience, Institute of Experimental Psychology, University of Duesseldorf, Duesseldorf, Germany

    M. A. de Souza Silva & J. P. Huston

Authors
  1. L. A. Ruocco
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  2. C. Treno
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  3. U. A. Gironi Carnevale
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  4. C. Arra
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  5. G. Boatto
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  6. C. Pagano
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  7. A. Tino
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  8. M. Nieddu
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  9. M. Michel
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  10. I. Prikulis
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  11. E. Carboni
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  12. M. A. de Souza Silva
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  13. J. P. Huston
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  14. A. G. Sadile
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  15. C. Korth
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Correspondence to A. G. Sadile.

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A. G. Sadile and C. Korth share the last authorship position.

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Ruocco, L.A., Treno, C., Gironi Carnevale, U.A. et al. Immunization with DISC1 protein in an animal model of ADHD influences behavior and excitatory amino acids in prefrontal cortex and striatum. Amino Acids 47, 637–650 (2015). https://doi.org/10.1007/s00726-014-1897-6

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