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P2–043: Combined nutrient supplementation enhances neurite outgrowth and synaptic protein expression in vitro
Author(s) -
Savelkoul Paul,
Lotstra Susanne,
Kuipers Almar,
Kamphuis Patrick,
Broersen Laus
Publication year - 2013
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2013.05.685
Subject(s) - neurite , synaptogenesis , synapse , microbiology and biotechnology , biology , dendritic spine , chemistry , biochemistry , in vitro , neuroscience , hippocampal formation
molecular weight soluble diffusible forms of Aß (mono/di/tetra/oligomers) are toxic. Very few studies have addressed identification and distribution of diverse Aß multimers in the AD brain or in the brains of AD-models. This investigation identified 4G8-immunoreactive (IR) and 6E10-IR Aß multimers in 5XFAD brains and tested if intranasal immunization using 3A1 antibody reduced any of these specific Aß multimers.Methods: 2-month-old 5XFAD mice (Tg) were intranasally immunized with 3A1 antibody (mAb provided by Drs. O’Nuallain and Lemere) for 8 weeks: Mice were administered with a total dose of 40mg/mouse/week [(5mg/5ul/naris)1⁄4(10mg/10ul/once), administered twice/day (1⁄420mg/day; twice/ week1⁄440mg/mouse/week], for 8 weeks (Immunized Tgs). Age-matched untreated Tgs received equal volume of sterile saline (Control Tgs). At the end, mice were euthanized, left brain hemisphere was processed for Western blot analysis for 6E10 and 4G8 immunoreactivity (IR); and right brain hemisphere was processed for 6E10 immunocytochemistry using established procedures. Quantitation of 4G8and 6E10-IR bands was performed using Kodak X-5100R densitometer. Quantitation of Aß plaque-associated 6E10-IR in brain sections was performed as established. Data were analyzed by ANOVA, Tukey posthoc test. Results: Immunoblotting with 4G8 showed that 2-month-old control Tgs at the beginning of treatment revealed the presence of only hexa/octo-deca/12mer-oligomers. 4-month-old control Tgs revealed additional detection of tetra/16-mers/dodecamers in addition to hexa/octo-deca/12mer-oligomers. Immunized Tgs showed reductions in 4G8-IR Aß multimers [tetramers (Y78.9%), hexamers (Y29.35%), octo-decamers (Y16.7%), 12-mers (Y35.35%), 16-mers (Y18.15%), dodecamers (Y23.3%)].Immunoblotting with 6E10 showed that 2-month-old control Tgs at the beginning of treatment revealed the presence of only hexa/octo-deca/12-mers-oligomers/dodecamers. 4-month-old control Tgs revealed additional detection of tetramers in addition to hexa/octo-deca/12-mers-oligomers/dodecamers. Immunized Tgs showed reduced 6E10-IR Aß multimers [tetramers (Y10.5%), hexamers (Y25.6%), octo-decamers (Y35%), 12-mers (Y26.1%), and dodecamers (Y34.34%)]. Immunized Tgs showed w40% reduction of 6E10-IR cortical plaques. Conclusions: Intranasal immunization with 3A1 antibody reduced the immunoexpression of all Aß multimers, with most prominent reductions in 4G8-IR dimers, tetramers and oligomers and 6E10-IR octo-decamers. The results will enhance development of new therapies for AD.