Abstract：
BACKGROUND:Primary cell culture is a valuable tool to utilize in parallel with in vivo studies in order to maximize our understanding of the mechanisms surrounding neurogenesis and central nervous system (CNS) regeneration and plasticity. The zebrafish is an important model for biomedical research and primary neural cells are readily obtainable from their embryonic stages viatissue dissociation. Further, transgenic reporter lines with cell type-specific expression allows for observation of distinct cell populations within the dissociated tissue. NEW METHOD:Here, we define an efficient method for ex vivo quantification and characterization of neuronal and glial tissue dissociated from embryonic zebrafish. RESULTS:Zebrafish brain dissociated cells have been documented to survive in culture for at least 9 days in vitro (div). Anti-HuC/D and anti-Acetylated Tubulin antibodies were used to identify neurons in culture; at 3 div approximately 48% of cells were HuC/D positive and 85% expressed serotonin, suggesting our protocol can efficiently isolate neurons from whole embryonic zebrafish brains. Live time-lapse imaging was also carried out to analyze cell migration in vitro. COMPARISON WITH EXISTING METHODS:Primary cultures of zebrafish neural cells typically have low rates of survivability in vitro. We have developed a culture system that has long term cell viability, enabling direct analysis of cell-cell and cell-extracellular matrix interactions. CONCLUSIONS:These results demonstrate a practical method for isolating, dissociating and culturing of embryonic zebrafish neural tissue. This approach could further be utilized to better understand zebrafish regeneration in vitro.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Patel BB,Clark KL,Kozik EM,Dash L,Kuhlman JA,Sakaguchi DSdoi
10.1016/j.jneumeth.2019.108419subject
Has Abstractpub_date
2019-12-01 00:00:00pages
108419eissn
0165-0270issn
1872-678Xpii
S0165-0270(19)30276-6journal_volume
328pub_type
杂志文章abstract：:Previous research has shown that spatio-temporal waves in the EEG are generally of long spatial wavelength and form smooth patterns of phase gradients at particular time-samples. This paper describes a method to measure smooth phase gradients of long spatial wavelength in the EEG. The method depends on the global patt...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
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abstract：BACKGROUND:Deep brain stimulation (DBS) in experimental animals has promoted new indications and refined existing treatments. Implantation of downscaled clinical DBS leads directly compatible with commercially available implantable pulse generators can however be challenging. Accordingly, we have developed a lead impla...
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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