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Changes of several brain receptor complexes in the cerebral cortex of patients with Alzheimer disease: probable new potential pharmaceutical targets

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Abstract

Although Alzheimer disease (AD) has been linked to defects in major brain receptors, studies thus far have been limited to the determination of receptor subunits or specific ligand binding studies. However, the availability of current technology enables the determination and quantification of brain receptor complexes. Thus, we examined levels of native receptor complexes in the brains of patients with AD. Cortical tissue was obtained from control subjects (n = 12 females and 12 males) and patients with AD (n = 12 females and 12 males) within a 3-h postmortem time period. The tissues were kept frozen until further biochemical analyses. Membrane proteins were extracted and subsequently enriched by ultracentrifugation using a sucrose gradient. Membrane proteins were then electrophoresed onto native gels and immunoblotted using antibodies against individual brain receptors. We found that the levels were comparable for complexes containing GluR2, GluR3 and GluR4 as well as 5-HT1A. Moreover, the levels of complexes containing muscarinic AChR M1, NR1 and GluR1 were significantly increased in male patients with AD. Nicotinic AChRs 4 and 7 as well as dopaminergic receptors D1 and D2 were also increased in males and females with AD. These findings reveal a pattern of altered receptor complex levels that may contribute to the deterioration of the concerted activity of these receptors and thus result in cognitive deficits observed in patients with AD. It should be emphasised that receptor complexes function as working units rather than individual subunits. Thus, the receptor deficits identified may be relevant for the design of experimental therapies. Therefore, specific pharmacological modulation of these receptors is within the pharmaceutical repertoire.

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Acknowledgments

We are grateful to the Banner Sun Health Research Institute Brain Donation Program of Sun City, Arizona, for the provision of human brain tissue. The Brain Donation Program is supported by the National Institute on Aging (P30 AG19610 Arizona Alzheimer’s Disease Core Center), the Arizona Department of Health Services (contract 211002, Arizona Alzheimer’s Research Center), the Arizona Biomedical Research Commission (contracts 4001, 0011, 05-901 and 1001 to the Arizona Parkinson’s Disease Consortium) and the Michael J. Fox Foundation for Parkinson’s Research.

Conflict of interest

The authors declare that there is no conflict of interest.

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Authors and Affiliations

  1. Department of Pediatrics, Medical University of Vienna, Währinger Gürtel 18–20, 1090, Vienna, Austria

    Soheil Keihan Falsafi, Maryam Ghafari & Gert Lubec

  2. Paul Flechsig Institute for Brain Research, University of Leipzig, Jahnallee 59, 04109, Leipzig, Germany

    Steffen Roßner & Markus Morawski

  3. Institute of Analytical Chemistry, University of Vienna, Waehringer Str. 38, 1090, Vienna, Austria

    Michael Groessl & Christopher Gerner

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  1. Soheil Keihan Falsafi
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Correspondence to Gert Lubec.

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S. Keihan Falsafi, S. Roßner, M. Ghafari are qually shared authorship.

Electronic supplementary material

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726_2013_1623_MOESM1_ESM.ppt

Supplemental Fig. 1 Arbitrary units of the optical density of receptor complexes were not significantly different between control subjects and patients with AD (PPT 13252 kb)

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Supplemental Fig. 2 Arbitrary units of the optical density of receptor complexes were not significantly different between control subjects and patients with AD (PPT 13302 kb)

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Supplemental Fig. 3 Arbitrary units of the optical density of receptor complexes were not significantly different between control subjects and patients with AD (PPT 5657 kb)

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Supplemental Fig. 4 Arbitrary units of the optical density of receptor complexes were not significantly different between control subjects and patients with AD (PPT 19356 kb)

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Supplemental Fig. 5 SDS-PAGE of the immunoprecipitate using the D1 receptor antibody. The D1 receptor and a single band containing NR1 as well as GluR1, 2 and 4 were identified by mass spectrometry (PPT 531 kb)

Supplemental Fig. 6 Representative mass spectra identifying D1, NR1, GluR1, 2 and 4 (PPT 522 kb)

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Supplemental Table 1 Representative Case numbers, gender, age and Braak scores of samples used for the analyses (DOC 119 kb)

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Supplemental Table 2 (A) A full list of the associations among the receptor complexes in male (B) and female control subjects (PPT 121 kb)

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Supplemental Table 3 (A) A full list of the associations among the receptor complexes in male and (B) female patients with AD (PPT 123 kb)

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Supplemental Table 4 A list of receptor peptides resulting from the in-gel digestion of the immunoprecipitate on the SDS-gel followed by mass spectrometrical analysis (XLS 41 kb)

Supplemental Table 5 Sequence coverage obtained by digestion with trypsin and chymotrypsin (PPT 70 kb)

Supplementary material 12(1a,b,c) (DOC 40 kb)

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Keihan Falsafi, S., Roßner, S., Ghafari, M. et al. Changes of several brain receptor complexes in the cerebral cortex of patients with Alzheimer disease: probable new potential pharmaceutical targets. Amino Acids 46, 223–233 (2014). https://doi.org/10.1007/s00726-013-1623-9

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