The isolation and culture of microglia-like cells from the goldfish brain

Houalla, Tarek; Levine, Robert L.

doi:10.1016/j.jneumeth.2003.08.004

Cited by 6 publications

(5 citation statements)

References 85 publications

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“…To develop the protocol for goldfish RGC culture system, we combined different primary glial cell culture protocols of mammalian astrocytes, fish astroglia, goldfish microglial cells, and zebrafish brainstem cells (Houalla and Levine, 2003 ; Crocker et al, 2008 ; Mack and Tiedemann, 2013 ; Tapanes-Castillo et al, 2014 ). There were several crucial differences between the protocols previously described and ours in order to optimize the isolation of RGCs.…”

Section: Discussionmentioning

confidence: 99%

“…Here a relatively higher percentage (15%) of FBS was used to increase the glial cell attachment to the dish and inhibit neuronal cell survival and growth, because certain components in serum, like glutamate, are toxic to neurons (Ye and Sontheimer, 1998 ). Furthermore, microglia and oligodendrocytes were removed and separated from the culture because of their differential adhesive properties compared to RGCs, as they need suitable cell culture medium supplements and take relatively longer time to adhere (Houalla and Levine, 2003 ). Oligodendrocytes and oligodendrocyte precursor cells are the least adhesive cell population in the brain, and can therefore be easily removed during changing of the cell culture medium (Itoh, 2002 ).…”

Section: Discussionmentioning

confidence: 99%

“…Oligodendrocytes and oligodendrocyte precursor cells are the least adhesive cell population in the brain, and can therefore be easily removed during changing of the cell culture medium (Itoh, 2002 ). According to Houalla et al, trypsin (0.25%) is toxic to goldfish microglial cells during the tissue dissociation step (Houalla and Levine, 2003 ), which was an advantage to our RGC culture development. However, trypsin solely is not enough for separating microglia from RGCs since in the first two passages some microglia-like cells appear on top of the RGC cell layer.…”

Section: Discussionmentioning

confidence: 99%

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Dopamine D1 receptor activation regulates the expression of the estrogen synthesis gene aromatase B in radial glial cells

et al. 2015

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Radial glial cells (RGCs) are abundant stem-like non-neuronal progenitors that are important for adult neurogenesis and brain repair, yet little is known about their regulation by neurotransmitters. Here we provide evidence for neuronal-glial interactions via a novel role for dopamine to stimulate RGC function. Goldfish were chosen as the model organism due to the abundance of RGCs and regenerative abilities of the adult central nervous system. A close anatomical relationship was observed between tyrosine hydroxylase-positive catecholaminergic cell bodies and axons and dopamine-D1 receptor expressing RGCs along the ventricular surface of telencephalon, a site of active neurogenesis. A primary cell culture model was established and immunofluorescence analysis indicates that in vitro RGCs from female goldfish retain their major characteristics in vivo, including expression of glial fibrillary acidic protein and brain lipid binding protein. The estrogen synthesis enzyme aromatase B is exclusively found in RGCs, but this is lost as cells differentiate to neurons and other glial types in adult teleost brain. Pharmacological experiments using the cultured RGCs established that specific activation of dopamine D1 receptors up-regulates aromatase B mRNA through a cyclic adenosine monophosphate-dependent molecular mechanism. These data indicate that dopamine enhances the steroidogenic function of this neuronal progenitor cell.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Dopamine D1 receptor activation regulates the expression of the estrogen synthesis gene aromatase B in radial glial cells

et al. 2015

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“…Microglia can be obtained from mixed cortical glial cultures originating from embryonic/fetal, adult, or diseased CNS of different species (rat [35]; swine [160]; human [266]; goldfish [347]; quail [348]). These cultures are started as a monolayer, mainly consisting of astrocytes with few contaminating fibroblasts, endothelial cells, and oligodendrocytes.…”

Section: Experimental Models For Microglia Research In Vitro and In Vivomentioning

confidence: 99%

Microglia: gatekeepers of central nervous system immunology

Tambuyzer

Ponsaerts

Nouwen

2008

Journal of Leukocyte Biology

290

246

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Microglia are perhaps the most underestimated cell type of our immune system. Not only were immunologists unaware of their capabilities until recently, but also, some neuroscientists denied their actual existence until the late 20th century. Nowadays, their presence is confirmed extensively, as demonstrated by numerous reports describing their involvement in virtually all neuropathologies. However, despite distinct approaches, their origin remains a point of controversy. Although many agree about their myeloid-monocytic ancestry, the precise progenitor cells and the differentiation mechanisms, which give rise to microglia in the different developmental stages of the CNS, are not unraveled yet. Mostly, this can be attributed to their versatile phenotype. Indeed, microglia show a high morphological plasticity, which is related to their functional state. This review about microglia aims to introduce the reader extensively into their ontogeny, cell biology, and involvement in different neuropathologies.

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“…In zebrafish, the movement of macrophages into the brain is a CSF-1R dependent process [13]. Microglia from goldfish brain tissue have been identified and shown to be capable of phagocytosis [33]. However, while it seems likely that CSF-1R would be involved in goldfish microglia development, further studies are required.…”

Section: Discussionmentioning

confidence: 99%