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. 2009 Oct 15;240(2):245-54.
doi: 10.1016/j.taap.2009.05.024. Epub 2009 Jun 6.

Increased beta-amyloid levels in the choroid plexus following lead exposure and the involvement of low-density lipoprotein receptor protein-1

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Increased beta-amyloid levels in the choroid plexus following lead exposure and the involvement of low-density lipoprotein receptor protein-1

Mamta Behl et al. Toxicol Appl Pharmacol. .

Abstract

The choroid plexus, a barrier between the blood and cerebrospinal fluid (CSF), is known to accumulate lead (Pb) and also possibly function to maintain brain's homeostasis of Abeta, an important peptide in the etiology of Alzheimer's disease. This study was designed to investigate if Pb exposure altered Abeta levels at the blood-CSF barrier in the choroid plexus. Rats received ip injection of 27 mg Pb/kg. Twenty-four hours later, a FAM-labeled Abeta (200 pmol) was infused into the lateral ventricle and the plexus tissues were removed to quantify Abeta accumulation. Results revealed a significant increase in intracellular Abeta accumulation in the Pb-exposed animals compared to controls (p<0.001). When choroidal epithelial Z310 cells were treated with 10 microM Pb for 24 h and 48 h, Abeta (2 microM in culture medium) accumulation was significantly increased by 1.5 fold (p<0.05) and 1.8 fold (p<0.05), respectively. To explore the mechanism, we examined the effect of Pb on low-density lipoprotein receptor protein-1 (LRP1), an intracellular Abeta transport protein. Following acute Pb exposure with the aforementioned dose regimen, levels of LRP1 mRNA and proteins in the choroid plexus were decreased by 35% (p<0.05) and 31.8% (p<0.05), respectively, in comparison to those of controls. In Z310 cells exposed to 10 microM Pb for 24 h and 48 h, a 33.1% and 33.4% decrease in the protein expression of LRP1 was observed (p<0.05), respectively. Knocking down LRP1 resulted in even more substantial increases of cellular accumulation of Abeta, from 31% in cells without knockdown to 72% in cells with LRP1 knockdown (p<0.05). Taken together, these results suggest that the acute exposure to Pb results in an increased accumulation of intracellular Abeta in the choroid plexus; the effect appears to be mediated, at least in part, via suppression of LRP1 production following Pb exposure.

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Figures

Fig. 1
Fig. 1
Increased accumulation of intracellular Aβ in rat choroid plexus tissues following in vivo acute Pb exposure by confocal study. (A). Choroid plexus tissue from a control rat. (B). Rats received a single ip injection of 27 mg Pb/kg. Twenty-four hr post exposure, FAM-labeled Aβ1–40 was infused into brain ventricles for 0.5 min. The plexus tissues were then removed 20 min post infusion for the confocal study. Substantial stains were evident in the cytosol but not in nuclei of choroidal epithelia. The lower panel shows the corresponding transmission image, indicating normal morphology of plexus tissues. (C). Quantification of the fluorescent signals using Laser Scanning Cytometry. Data represent mean ± SD, n=16 (a total of 16 cells per group taken from 4 tissue samples with fluorescence averaged from 4 cells per sample). **: p<0.001 as compared to controls.
Fig. 2
Fig. 2
Increased accumulation of Aβ in immortalized Z310 cells following in vitro Pb exposure by confocal study. (A). Control cells. (B). Cells were incubated with 10 uM Pb for 24 hr. (C). Cells were incubated with 10 μM Pb for 48 hr. An increase in intracellular Aβ signals was evident in Pb-exposed cells as compared to controls. The lower panel shows the corresponding transmission image, indicating a normal morphology of Z310 cells.
Fig. 3
Fig. 3
Increases in accumulation of intracellular Aβ in Z310 cells following Pb exposure as quantified by ELISA. (A). Dose-response study. Cells were treated with Pb at the concentrations indicated for 24 hr. The cells were harvested and homogenates used for ELSIA. The value in parenthesis was excluded from the linear regression analysis. (B). Time-course study. Cells were treated with 10 μM for 4–24 h, followed by ELISA. Data represent mean ± SD, n = 4–6 wells per group. *: p<0.05.
Fig. 4
Fig. 4
Decreased LRP1 mRNA expression following in vivo or in vitro Pb exposure. (A). Rats received ip injection of either Na-acetate (control) or Pb acetate (27 mg Pb/kg) and tissues were analyzed 24h after Pb exposure. (B). Z310 cells were treated with 10 μM Pb for 24h and 48 h. The relative mRNA levels of LRP1 and GAPDH were quantified by real-time RT-PCR and expressed as the ratio of LRP1/GAPDH. Data represent mean ± SD, n=4; *: p<0.05 as compared to control. The data are representative of triplicate experiments.
Fig. 5
Fig. 5
Decreased LRP1 protein expression following in vivo or in vitro Pb exposure by Western blot analysis. (A) and (B): In vivo study. Rats received ip injection of either Na-acetate (control) or Pb acetate (27 mg Pb/kg) and tissues were analyzed 24h after Pb exposure. Data presented in (B) were estimated from the corresponding band densities in (A) and normalized to those of β-actin. (C) and (D): In vitro study. Z310 cells were treated with 10 μM Pb for 24h or 48 h. Data presented in (D) were estimated from the corresponding band densities in (C) and normalized to those of β-actin. Data represent mean ± SD, n=4; p<0.05 compared to controls.
Fig. 6
Fig. 6
Optimization of LRP1 siRNA. Z310 cells were cultured with 50 nM siRNA for 48h and LRP1 knockdown was verified by three independent methods. (A). Representative bands from a Western Blot indicating a 52% knockdown in LRP1 protein expression compared to controls. (B). Reduced LRP1 mRNA expression by real-time RT-PCR analysis. There was a 53% knockdown in LRP1 mRNA. (C). Representative image from Laser scanning cytometry analysis. (D). Quantitative fluorescence intensity. LRP1(−) indicates LRP1 knockdown and control indicates scrambled siRNA There was a 50% reduction in LRP1-related fluorescent signal in the LRP1(−) group compared to controls. No evident difference in cell confluence was observed between the control and LRP1-knockdown groups. Data represent mean ± SD, n=4–6; *: p<0.05 as compared to controls. The data are representative of triplicate experiments.
Fig. 7
Fig. 7
Quantification of Aβ concentrations by ELISA. A plus sign (+) for LRP1(−) (knockdown) indicates the cells with LRP1 knockdown; a minus sign (−) for LRP1(−) indicates the control cells with scrambled siRNA. Following recovery from the transfection of LRP1 or scrambled siRNA, cells were incubated with the media in presence of 10 μM Pb (with a plus sign) or in absence of Pb (with a minus sign) for 24 h, followed by incubation with 2 μM Aβ for 1 h. Data represent mean ± SD, n = 4–6. Bars with different superscripts are significantly different from one another, p<0.05.

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References

    1. Atkins DS, Basha MR, Zawia NH. Intracellular signaling pathways involved in mediating the effects of lead on the transcription factor Sp1. International Journal of Developmental Neuroscience. 2003;21:235–244. - PubMed
    1. Basha MR, Wei W, Bakheet SA, Benitez N, Siddiqi HK, Ge YW, Lahiri DK, Zawia NH. The fetal basis of amyloidogenesis: exposure to lead and latent overexpression of amyloid precursor protein and beta-amyloid in the aging brain. J Neurosci. 2005a;25:823–9. - PMC - PubMed
    1. Basha MR, Murali M, Siddiqi HK, Ghosal K, Siddiqi OK, Lashuel HA, Ge YW, Lahiri DK, Zawia NH. Lead (Pb) exposure and its effect on APP proteolysis and Aβ aggregation. The FASEB Journal express article. 2005b;10:1–16. - PubMed
    1. Bell RD, Sagare AP, Friedman AE, Bedi GS, Holtzman DM, Deane R, Zlokovic BV. Transport pathways for clearance of human Alzheimer’s amyloid β-peptide and apolipoproteins E and J in the mouse central nervous system. Journal of Cerebral Blood Flow & Metabolism. 2007;27:909–918. - PMC - PubMed
    1. Brody DL, Magnoni S, Schwetye KE, Spinner ML, Esparza TJ, Stocchetti N, Zipfel GJ, Holtzman DM. Amyloid-β dynamics correlate with neurological status in the injured human brain. Science. 2008;321:1221–1224. - PMC - PubMed

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