CC
Cristina Carvalho
Research fields
  • Neuroscience
Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus
Authors:  Valeria Lallai, Amina Ahmed and Christie D. Fowler, date: 02/20/2020, view: 4505, Q&A: 0
The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular environment. This biofluid contains an enriched source of proteins, ions, and other signaling molecules for extracellular support of neurons and glial cells within the central nervous system. Given that other cells in the brain also release factors into the CSF, in vitro investigations of choroid plexus function are necessary to isolate processes selectively occurring within and released from this tissue. Here, we describe a protocol to isolate choroid plexus tissue from each of the ventricular locations, and the cell culture conditions required to support growth and maintenance of these epithelial cells. This technique allows for investigations of the functional significance of the choroid plexus, such as for the examination of stimuli promoting the release of growth factors and extracellular vesicles (e.g., exosomes and microvesicles) from ventricle-specific choroid plexus epithelial cells.
Evaluation of Genotoxicity by Micronucleus Assay in vitro and by Allium cepa Test in vivo
Authors:  Christina N. Banti and Sotiris K. Hadjikakou, date: 07/20/2019, view: 7923, Q&A: 0
The in vitro and in vivo genotoxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) is evaluated by the micronucleus test and the Allium cepa assay, respectively. Fetal lung fibroblast cells (MRC-5), normal human corneal epithelial cells (HCEC) and immortalized human keratinocytes cells (HaCaT) were incubated with solutions of SBAMs or CoMeDs at their IC50 values for 48 h (the concentration of a compound which is required to inhibit the cells growth by 50% in relation to the non-treated cells). The micronucleus abundance percentage towards the corresponding one, of the non-treated cells indicates the in vitro genotoxicity of the formulations. The in vivo Allium cepa test comprises the exposing of the plant Allium cepa roots to an SBAMs or a CoMeDs solution for 48 h. The percentages of the mitotic index, the chromosome aberrations, the nuclear abnormalities and the presence of the micronucleus are calculated indicating the in vivo genotoxicity of the agent.
Social Defeat Stress (SDS) in Mice: Using Swiss Mice as Resident
Authors:  Marco Oreste F. Pagliusi Jr. and Cesar R. Sartori, date: 03/20/2019, view: 5682, Q&A: 0
Due to the high prevalence and great economic impact of depression, studies with animal models have been increasingly used to identify neurobiological mechanisms associated with this disorder. However, many animal models use stressful conditions that are not consistent with what we observe in the modern human world. Examples are the chronic unpredictable stress and the electric shock model used in rodents. It’s well established the social stress as the major cause of depressive disorder in human, in this way a social defeat stress model was recently standardized and can induce depressive-like behavior of social avoidance, a typical human depressive behavior. In this model, mice are exposed on consecutive days to an aggressor mouse, suffering brief periods of physical aggression followed by longer periods of visual and olfactory (sensory) contact and, as a consequence, a relationship of social submission is characterized. Thus, the objective of this work is to describe a social defeat stress protocol using swiss mice as resident, also describing valuable procedural suggestions that will help researchers to reproduce the model easily.
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