Experimental Tools to Study the Regulation and Function of the Choroid Plexus

Gonçalves, Isabel; Quintela, Telma; Duarte, Ana Catarina; Hubbard, Peter C.; Baltazar, Graça; Schwerk, Christian; Belin, Andrea Carmine; Tomás, Joana; Santos, Cecília R. A.

doi:10.1007/978-1-4939-8946-1_13

Cited by 3 publications

(2 citation statements)

References 53 publications

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“…For Aβ uptake assays, HIBCPP cells were plated in culture inserts (pore diameter .4 μm, .33 cm 2 ; VWR, Portugal) as previously described (Gonçalves et al, 2019; Schwerk et al, 2012). In brief, HIBCPP cells were seeded in the upper chamber of the insert at a density of 1.5 × 10 5 using a culture medium supplemented with 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

Circadian rhythmicity of amyloid‐beta‐related molecules is disrupted in the choroid plexus of a female Alzheimer's disease mouse model

Furtado

Esgalhado

Duarte

et al. 2022

J of Neuroscience Research

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The choroid plexus (CP) is part of the blood‐cerebrospinal fluid barrier (BCSFB) and was recently described as an important component of the circadian clock system. It is the principal source of cerebrospinal fluid (CSF) and responsible for the synthesis and secretion of various neuroprotective peptides including those involved in amyloid‐β (Aβ) transport/degradation, contributing to Aβ homeostasis. Inadequate Aβ metabolic clearance and transport across the BCSFB have been associated with circadian dysfunctions in Alzheimer's disease (AD) patients. To investigate whether AD pathology influences Aβ scavengers circadian expression, we collected CP at different time points from an AD mouse model (APP/PS1) (female and male animals, aged 6‐ and 12‐months‐old) and analyzed their mRNA expression by Real‐time RT‐PCR. Only angiotensin‐converting enzyme (Ace) expression in 6‐month‐old female wild‐type mice and transthyretin (Ttr) expression in 12‐month‐old female wild‐type mice presented significant rhythmicity. The circadian rhythmicity of Ace and Ttr, prompt us to analyze the involvement of circadian rhythm in Aβ uptake. A human CP papilloma (HIBCPP) cell line was incubated with Aβ‐488 and uptake was evaluated at different time points using flow cytometry. Aβ uptake displayed circadian rhythmicity. Our results suggest that AD might affect Aβ scavengers rhythmicity and that Aβ clearance is a rhythmic process possibly regulated by the rhythmic expression of Aβ scavengers.

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Section: Methodsmentioning

confidence: 99%

Circadian rhythmicity of amyloid‐beta‐related molecules is disrupted in the choroid plexus of a female Alzheimer's disease mouse model

Furtado

Esgalhado

Duarte

et al. 2022

J of Neuroscience Research

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“…Thirty postnatal animals were anesthetized on ice for at least 30 min before being euthanized. The CPs were collected from the lateral ventricles and used to establish choroid plexus epithelial cell (CPEC) primary cultures as previously described by Gonçalves et al [39]. Briefly, the dissociated cells were seeded into 6-well culture plates and cultured in a high-glucose DMEM supplemented with5 µg/mL insulin (Sigma-Aldrich, Merck, Algés, Portugal), 100 U/mL penicillin, 100 µg/mL streptomycin, 10% v/v fetal bovine serum (FBS), 10 ng/mL epidermal growth factor (Sigma-Aldrich, Merck, Portugal) and 30 µM cytosine arabinoside (Sigma-Aldrich, Merck, Portugal).…”

Section: Choroid Plexus Epithelial Primary Culturementioning

confidence: 99%

Circadian ABCG2 Expression Influences the Brain Uptake of Donepezil across the Blood–Cerebrospinal Fluid Barrier

Furtado,

Duarte,

Costa

et al. 2024

IJMS

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Donepezil (DNPZ) is a cholinesterase inhibitor used for the management of Alzheimer’s disease (AD) and is dependent on membrane transporters such as ABCG2 to actively cross brain barriers and reach its target site of action in the brain. Located in the brain ventricles, the choroid plexus (CP) forms an interface between the cerebrospinal fluid (CSF) and the bloodstream, known as the blood–CSF barrier (BCSFB). Historically, the BCSFB has received little attention as a potential pathway for drug delivery to the central nervous system (CNS). Nonetheless, this barrier is presently viewed as a dynamic transport interface that limits the traffic of molecules into and out of the CNS through the presence of membrane transporters, with parallel activity with the BBB. The localization and expression of drug transporters in brain barriers represent a huge obstacle for drug delivery to the brain and a major challenge for the development of therapeutic approaches to CNS disorders. The widespread interest in understanding how circadian clocks modulate many processes that define drug delivery in order to predict the variability in drug safety and efficacy is the next bridge to improve effective treatment. In this context, this study aims at characterizing the circadian expression of ABCG2 and DNPZ circadian transport profile using an in vitro model of the BCSFB. We found that ABCG2 displays a circadian pattern and DNPZ is transported in a circadian way across this barrier. This study will strongly impact on the capacity to modulate the BCSFB in order to control the penetration of DNPZ into the brain and improve therapeutic strategies for the treatment of AD according to the time of the day.

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