Journal : IJNPPY Article Doi
: 10.1093/ijnp/pyw051
Article Title
: Reduced Neurite Density in Neuronal Cell Cultures Exposed to Serum of Patients with Bipolar Disorder
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AUTHOR QUERY FORM
Journal : IJNPPY Article Doi
: 10.1093/ijnp/pyw051
Article Title : Reduced Neurite Density in Neuronal Cell Cultures Exposed to Serum of Patients with Bipolar Disorder First Author : Bianca Wollenhaupt-Aguiar Corr. Author : Bianca Wollenhaupt-Aguiar AUTHOR QUERIES - TO BE ANSWERED BY THE CORRESPONDING AUTHOR The following queries have arisen during the typesetting of your manuscript. Please click on each query number and respond by indicating the change required within the text of the article. If no change is needed please add a note saying “No change.” AQ1 AQ2 AQ3 AQ4 AQ5 AQ6 AQ7 AQ8 AQ9
Please provide expansion for BDNF. Please revise this sentence for clarity. Please provide expansion for NIS. Please provide expansion for DMEM. Please revise sentence for clarity. Please expand FKL: no author name matches these initials. Please expand FKL: no author name matches these initials. Please clarify whether this is Dr Kapczinski or Klamt. Please provide all author names unless there are more than 20.
Copyedited by: OUP
International Journal of Neuropsychopharmacology (2016) 00(00): 1–5 doi:10.1093/ijnp/pyw051 Advance Access Publication: XXXX XX, XXXX Rapid Communication
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Reduced Neurite Density in Neuronal Cell Cultures Exposed to Serum of Patients with Bipolar Disorder
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Bianca Wollenhaupt-Aguiar, MSc; Bianca Pfaffenseller, MSc; Vinicius de Saraiva Chagas, BSc; Mauro A A Castro, MD, PhD; Ives Cavalcante Passos, MD, PhD; Márcia Kauer-Sant’Anna, MD, PhD; Flavio Kapczinski, MD, PhD; Fábio Klamt, PhD
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Bipolar Disorder Program, Laboratory of Molecular Psychiatry, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil (Drs Wollenhaupt-Aguiar, Pfaffenseller, Passos, Kauer-Sant’Anna, and Kapczinski); Laboratory of Cellular Biochemistry, Post-Graduate Program in Biochemistry, Biochemistry Department, ICBS/ UFRGS, Porto Alegre, Brazil (Drs Wollenhaupt-Aguiar, Pfaffenseller, and Klamt); Bioinformatics and Systems Biology Laboratory, Post-Graduate Program in Bioinformatics, Federal University of Paraná (UFPR), Polytechnic Center, Curitiba, Brazil (Drs de Saraiva Chagas and Castro); Graduation Program in Psychiatry and Department of Psychiatry, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil (Drs Passos, KauerSant’Anna, and Kapczinski)
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Correspondence: Professor Fábio Klamt, PhD, Laboratory of Cellular Biochemistry, Federal University of Rio Grande do Sul, Avenida Ramiro Barcelos, 2600, Porto Alegre (RS) 90035–903 Brazil (
[email protected])
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Abstract
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Background: Increased inflammatory markers and oxidative stress have been reported in serum among patients with bipolar disorder (BD). The aim of this study is to assess whether biochemical changes in the serum of patients induces neurotoxicity in neuronal cell cultures. Methods: We challenged the retinoic acid-differentiated human neuroblastoma SH-SY5Y cells with the serum of BD patients at early and late stages of illness and assessed neurite density and cell viability as neurotoxic endpoints. Results: Decreased neurite density was found in neurons treated with the serum of patients, mostly patients at late stages of illness. Also, neurons challenged with the serum of late-stage patients showed a significant decrease in cell viability. Conclusions: Our findings showed that the serum of patients with bipolar disorder induced a decrease in neurite density and cell viability in neuronal cultures.
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Keywords: bipolar disorder, neurite density, RA-differentiated SH-SY5Y cells, systemic toxicity 95 45
Introduction 50
Bipolar disorder (BD) affects about 2% of the world’s population, with sub-threshold forms affecting up to a further 2% (Merikangas et al., 2007). The course of BD is highly variable, and a subset of patients seem to present a progressive course
associated with brain changes (Cao et al., 2016) and functional impairment (Rosa et al., 2014). Nonetheless, the molecular foundations for this illness progression are just beginning to be explained. It is known that BDNF serum levels were decreased in
Received: April 13, 2016; Revised: May 7, 2016; Accepted: May 9, 2016 © The Author 2016. Published by Oxford University Press on behalf of CINP. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http:// creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact
[email protected]
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the late-stage of BD when compared to those at an early stage of the illness (Kauer-Sant’Anna et al., 2009). Moreover, altered cellular resilience was reported in late-stage patients (Pfaffenseller et al., 2014). In addition, patients at a late stage of illness present increased levels of C-C motif ligand 11 (CCL11) and decreased levels of CXCL827 chemokines (Panizzutti et al., 2015). However, what is not known is how these abnormal peripheral blood markers may lead to, or be involved with, brain changes in BD. In 2011, our group hypothesized that impairments in neuroplasticity of BD patients may be translated into shrinkage of the brain structures by reducing neurites and intercellular connections in the neuronal network (Berk et al., 2011). We also suggested that the aforementioned biochemical changes may play a causal role in this scenario, which has been called the systemic toxicity (Kapczinski et al., 2010). Recently, several studies reported reductions in the volume of the left hippocampi (Cao et al., 2016) and frontal cortices of patients with late-stage BD (Abe et al., 2015). These findings are in line with the pioneering work of Strakowski and colleagues (2002), which reports increased ventricle volumes in multiple-episode patients with BD compared to those who had only one episode. However, the causal role of the abnormal peripheral blood markers on neuronal cells of the patients with late-stage BD has not been investigated yet. In the present study, we used an in vitro approach with the retinoic acid (RA)-differentiated human cell line SH-SY5Y exposed to serum of bipolar patients. The differentiated human neuroblastoma cell line, SH-SY5Y, has been used as an experimental model to assess molecular and biochemical pathways involved in the pathophysiology of brain disorders (Lopes et al., 2010) and for neurotoxicological experiments in developmental and mature neurons (Schonhofen et al., 2015). This model has the advantage of being derived of human cells, displaying neuronal morphology, and neuronal markers (as high neurite density, tyrosine hydroxylase, dopamine transporter) during RA-differentiation (Lopes et al., 2010). Therefore, the aim of the present study was to assess whether biochemical changes in serum of patients with BD could induce neurotoxicity in neuronal cell cultures.
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Methods
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The Ethical Committee of the Hospital de Clínicas de Porto Alegre (HCPA) approved the study (application number: 12–0102). All subjects had signed the informed consent.
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DSM-IV axis-I Disorders (SCID-I) and Functioning Assessment Short Test (FAST), respectively, which were administered by trained staff. Current dimensional mood symptoms were assessed with the Hamilton Depression Rating Scale (HDRS; Hamilton, 1960) and the Young Mania Rating Scale (YMRS; Young et al., 1978). Euthymia was defined by the HDRS score < 8 and YMRS score
