Methodological factors influencing measurement and processing of plasma reelin in humans
Open accessAbstract
Background Reelin, intensively studied as an extracellular protein that regulates brain development, is also expressed in a variety of tissues and a circulating pool of reelin exists in adult mammals. Here we describe the methodological and biological foundation for carrying out and interpreting clinical studies of plasma reelin.Results Reelin in human plasma was sensitive to proteolysis, freeze-thawing and heating during long-term storage, sample preparation and electrophoresis. Reelin in plasma was a dimer under denaturing conditions. Boiling of samples resulted in laddering, suggesting that each of the 8 repeats expressed in reelin contains a heat-labile covalent bond susceptible to breakage. Urinary-type and tissue-type plasminogen activator converted reelin to a discrete 310 kDa fragment co-migrating with the major immunoreactive reelin fragment seen in plasma and also detected in brain. (In contrast, plasmin produced a spectrum of smaller unstable reelin fragments.) We examined archival plasma of 10 pairs of age-matched male individuals differing in repeat length of a CGG repeat polymorphism of the 5'-untranslated region of the reelin gene (both alleles < 11 repeats vs. one allele having >11 repeats). Reelin 310 kDa band content was lower in subjects having the long repeats in all 10 pairs, by 25% on average (p < 0.001). In contrast, no difference was noted for amyloid precursor protein.Conclusions Our studies indicate the need for caution in measuring reelin in archival blood samples, and suggest that assays of plasma reelin should take into account three dimensions that might vary independently: a) the total amount of reelin protein; b) the relative amounts of reelin vs. its proteolytic processing products; and c) the aggregation state of the native protein. Reelin-plasminogen activator interactions may affect their roles in synaptic plasticity. Our results also suggest that the human CGG repeat polymorphism affects reelin gene expression, and may affect susceptibility to human disease.
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Both Reelin protein levels and PON1 enzymatic activity are significantly reduced in AD individuals compared to controls.
“Both Reelin protein levels and PON1 enzymatic activity (measured as “arylesterase” in these studies) are significantly reduced in AD individuals compared to controls [89, 236–238].”
From Consensus Paper: Pathological Role of the Cerebellum in Autism (2012) · cites this paper as [238]
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Reelin, GABA, FMRP, and Autism
Folsom, T. D., & Fatemi, S. H. (2015) · Contemporary clinical neuroscience · doi:10.1007/978-1-4939-2190-4_16 · Cited by 1 · see citing works
Samama, B., & Boehm, N. (2008) · Springer eBooks · doi:10.1007/978-0-387-76761-1_17 · Cited by 1 · see citing works
Alzheimer’s Disease and Reelin
Botella-López, A., & Sáez‐Valero, J. (2008) · Springer eBooks · doi:10.1007/978-0-387-76761-1_26 · Cited by 1 · see citing works
- Authors
- Lugli, G., Krueger, J. M., Davis, J. M., Persico, A. M., Keller, F., & Smalheiser, N. R.
- Affiliations
- University of Illinois Chicago (US)
- Università Campus Bio-Medico (IT)
- Corresponding author
- Smalheiser, Neil R
- Venue
- BMC Biochemistry
- Published
- 2003-09-07
- Volume
- 4
- Issue
- 1
- Pages
- 9-9
- ISSN
- 1471-2091 (ISSN-L)
- PMC
- PMC200967
- OpenAlex
- W1789054710
- Cited by
- 60
- References
- 36
- Citations by year
- 2024: 1 · 2023: 1 · 2022: 3 · 2021: 1 · 2020: 3 · 2019: 1 · 2018: 2 · 2016: 5 · 2015: 2 · 2014: 1 · 2013: 5 · 2012: 6
- FWCI
- 2.11
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- BMC Biochemistry (publishedVersion)
- PubMed Central (publishedVersion, other-oa)
- PubMed (publishedVersion)
- IRIS UNIMORE (University of Modena and Reggio Emilia) (submittedVersion, cc-by)
- Funders
- National Institutes of Health (5r03mh060778-02)
- National Institute of Mental Health (MH60778)
- Topics
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
- Health Sciences › Medicine › Hematology › Blood Coagulation and Thrombosis Mechanisms
- Keywords
- plasminogen activator, plasmin, amyloid precursor protein, synaptic plasticity
- MeSH terms
- Reelin Protein, Calibration, Fibrinolysin, Hot Temperature, Humans, Male, Nerve Tissue Proteins, Plasminogen Activators, Serine Endopeptidases, Blotting, Western, Reproducibility of Results, Cell Adhesion Molecules, Neuronal, Extracellular Matrix Proteins, Dimerization, Repetitive Sequences, Amino Acid
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