Multilineage Differentiation Potential of CNS Cell Progenitors in a Recent Developed Gilthead Seabream (Sparus aurata L.) Nervous Model (original) (raw)
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Isolation and characterization of a neural progenitor cell line from tilapia brain
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2008
Astroglial cell lines have many applications for advancing neural developmental and functional studies. However, few astroglial cell lines have been reported from fish. In this study, we report the characterization of the immortal cell line TB2 isolated from adult tilapia brain tissue. The cell line was established at 25°C in L15 medium supplemented with 15% fetal bovine serum. Most of the cells displayed a fibrous morphology and were immunoreactive for A2B5 antigen, glial fibrillary acidic protein (GFAP), keratin, vimentin, and the gap junction protein connexin 43 (Cx43). They weakly expressed glutamine synthetase (GS), S100 protein, and the neural stem cell markers Sox2 and brain lipid binding protein (BLBP). In contrast to astroglia in vivo, most TB2 cells also expressed galactocerebroside (GalC), substance P (SP), and tyrosine hydroxylase (TH). By immunoblot and RT-PCR, the cells also expressed myelin basic protein (MBP), proteolipid protein (PLP), and Cx35. On a poly-L-lysine-coated substrate in vitro, TB2 cells showed increases in neuronal dopamine decarboxylase (DDC) and microtubule-associated protein 2 (MAP2), indicating that they can initiate differentiation into neurons. Taken together, the results suggest that TB2 cells are astroglial progenitor cells (neural stem cells) and may develop into oligodendrocytes and neurons in a suitable environment. The present study advances our knowledge of fish astroglia. However, the factors that affect neural development in fish remain unknown, as do the characteristics of the intermediate differentiation stages between stem cells and mature nerve cells. The TB2 cell line will promote these investigations.
Establishment of long term cultures of neural stem cells from adult sea bass, Dicentrarchus labrax
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2009
Long term cell cultures could be obtained from brains of adult sea bass (Dicentrarchus labrax) up to 5 days post mortem. On three different occasions, sea bass brain tissues were dissected, dispersed and cultured in Leibovitz's L-15 media supplemented with 10% fetal bovine serum. The resulting cellular preparations could be passaged within 2 or 3 weeks of growth. The neural cells derived from the first trial (SBB-W1) have now been passaged over 24 times within two years. These cells have been cryopreserved and thawed successfully. SBB-W1 cells are slow growing with doubling times requiring at least 7 days at 22°C. These long term cell cultures could be grown in suspension as neurospheres that were immunopositive for nestin, a marker for neural stem cells, or grown as adherent monolayers displaying both glial and neural morphologies. Immunostaining with anti-glial fibrillary acidic protein (a glial marker) and anti-neurofilament (a neuronal marker), yielded positive staining in most cells, suggesting their possible identity as neural stem cells. Furthermore, Sox 2, a marker for neural stem cells, could be detected from these cell extracts as well as proliferating cell nuclear antigen, a marker for proliferating cells. SBB-W1 could be transfected using pEGFP-N1 indicating their viability and suitability as convenient models for neurophysiological or neurotoxicological studies.
Italian Journal of Animal Science
As its Greek name suggests, neuroglia is the glue of the nervous system, simply referred to as non-neuronal cells, with supporting func-tions for neurons in the central nervous sys-tem (CNS) of mammalian and lower verte-brates. Recent advances in stem cell biology have redefined new functions. In fish, neu-roglial cells have been described as supporting cells, and as such require further investiga-tion. Approximately 283 cell lines have been obtained from fish worldwide, yet none from the brain of Sparus aurata, neither in cell lines nor as primary culture. We describe a novel reproducible in vitro neuroglial marine model for establishing primary neuroglial cell cul-tures by dissociating the whole brain of seabream juveniles. Proliferating neural stem cells produced alongside three generating lin-eages (neuronal precursor cells, astroglial pre-cursor cells and oligodendroglia precursor cells) developed neurons, astrocytes and oligo-dendrocytes, respectively. The radial glia, fine-ly...
Progenitor Radial Cells and Neurogenesis in Pejerrey Fish Forebrain
Brain, Behavior and Evolution, 2010
The central nervous system of adult teleost fish is peculiar because of the following features: (1) the persistence of radial glial cells, (2) an important neurogenic activity and (3) a high aromatase expression by radial cells. In this study, the proliferative zones of the forebrain were described using bromodeoxyuridine (BrdU) treatment in the brain of the pejerrey, an Acanthopterygian teleost fish. These cells were shown to have morphological and immunoreactive characteristics of radial cells and to express aromatase. Three different progenitor populations were identified based on the mobility and proliferation capacity 6 weeks after BrdU treatment: transit amplifying progenitors, slowly proliferating stem cells, and cells remaining in the proliferative zones showing no signs of mitotic activity. The proliferative cells were always located in the ventricular zone and were never observed in the brain parenchyma; however, 3 weeks later they were found away from these proliferative zones and displayed acetylated tubulin immunoreactivity. Other BrdU-positive cells showed astrocyte morphology and were immunoreactive to the S100 glial marker. These results show that in this fish, radial cells are true progenitors generating neurons and possibly astrocytes.
Neurogenesis and Neuronal Regeneration in the Adult Reptilian Brain
Brain Behavior and Evolution, 2001
Fish are distinctive in their enormous potential to continuously produce new neurons in the adult brain, whereas in mammals adult neurogenesis is restricted to the olfactory bulb and the hippocampus. In fish new neurons are not only generated in structures homologous to those two regions, but also in dozens of other brain areas. In some regions of the fish brain, such as the optic tectum, the new cells remain near the proliferation zones in the course of their further development. In others, as in most subdivisions of the cerebellum, they migrate, often guided by radial glial fibers, to specific target areas. Approximately 50% of the young cells undergo apoptotic cell death, whereas the others survive for the rest of the fish's life. A large number of the surviving cells differentiate into neurons. Two key factors enabling highly efficient brain repair in fish after injuries involve the elimination of damaged cells by apoptosis (instead of necrosis, the dominant type of cell death in mammals) and the replacement of cells lost to injury by newly generated ones. Proteome analysis has suggested well over 100 proteins, including two dozen identified ones, to be involved in the individual steps of this phenomenon of neuronal regeneration.
Isolation and long-term culture of neural stem cells from chondrostei fish Acipenser persicus
2018
In the present study, an in vitro brain cell culture was developed from neural cells of Persian sturgeon (Acipenser persicus). The tissue samples collected from the anterior, middle and posterior regions of the brain were cultivated separately in DMEM/F12 medium supplemented with 15% fetal bovine serum, antibiotic and antimycotic. The medium was refreshed every 3 days. The cells became confluent after about 3 weeks from the initial time of seeding. The cultured cells from the posterior part of the brain showed high potential of proliferation as they had been passaged 16 times in more than 11 months. To determine optimal temperature, the brain cells were incubated at four temperatures including; 20, 22, 25 and 28°C. The best cultivation temperature was obtained at 25°C. The cultured cells from posterior part of the brain were cryopreserved successfully and the survival rate was 70% after thawing. Immunocytochemistry using antibody against nesting showed that some cells were immunopositive for nesting. Finally, these results suggested that cell cultures from posterior part of the Persian sturgeon brain with high proliferation capacity can be useful for research on brain cells in A. persicus in the future.
Reviews in the Neurosciences, 2018
The review is an overview of the current knowledge of neuronal regeneration properties in mammals and fish. The ability to regenerate the damaged parts of the nervous tissue has been demonstrated in all vertebrates. Notably, fish and amphibians have the highest capacity for neurogenesis, whereas reptiles and birds are able to only regenerate specific regions of the brain, while mammals have reduced capacity for neurogenesis. Zebrafish (Danio rerio) is a promising model of study because lesions in the brain or complete cross-section of the spinal cord are followed by an effective neuro-regeneration that successfully restores the motor function. In the brain and the spinal cord of zebrafish, stem cell activity is always able to re-activate the molecular programs required for central nervous system regeneration. In mammals, traumatic brain injuries are followed by reduced neurogenesis and poor axonal regeneration, often insufficient to functionally restore the nervous tissue, while spi...
Neuronal stem cells in the brain of adult vertebrates
STEM CELLS, 1995
It is generally assumed that neurogenesis in the central nervous system ceases before or soon after birth. In the last three decades, however, several studies have reported that new neurons continue to be added into the brain of adult fish, frogs, reptiles, birds and mammals. The precursor cells that give rise to the neurons generated in adulthood are generally located in the walls of the brain ventricles. From these proliferative regions, neuronal precursors migrate toward their final targets where they differentiate; they often traverse long distances through complex brain parenchyma. The identity of the neuronal precursors in the brains of adult animals is still unknown. Experiments in adult birds suggest that proliferating radial cells may be the neuronal precursors. In adult mice, cells present in the subventricular zone can generate neurons in vivo and in vitro. These neuronal precursors can be induced to proliferate in vitro when exposed to growth factors and retain their potential to differentiate into neurons and glia. Whether these putative neural stem cells can differentiate into multiple neuronal types remains to be determined. The neuronal precursors of the adult brain could be used as a source of cells for neuronal transplantation. In addition, these cells could be manipulated in vivo or in vitro to introduce genes into the brain. Adult neurogenesis offers new experimental opportunities to study neuronal birth, migration and differentiation and for the treatment of neurological diseases.
Radial glial cells play a key role in echinoderm neural regeneration
BMC biology, 2013
"BACKGROUND: Unlike the mammalian central nervous system (CNS), the CNS of echinoderms is capable of fast and efficient regeneration following injury and constitutes one of the most promising model systems that can provide important insights into evolution of the cellular and molecular events involved in neural repair in deuterostomes. So far, the cellular mechanisms of neural regeneration in echinoderm remained obscure. In this study we show that radial glial cells are the main source of new cells in the regenerating radial nerve cord in these animals. RESULTS: We demonstrate that radial glial cells of the sea cucumber Holothuria glaberrima react to injury by dedifferentiation. Both glia and neurons undergo programmed cell death in the lesioned CNS, but it is the dedifferentiated glial subpopulation in the vicinity of the injury that accounts for the vast majority of cell divisions. Glial outgrowth leads to formation of a tubular scaffold at the growing tip, which is later populated by neural elements. Most importantly, radial glial cells themselves give rise to new neurons. At least some of the newly produced neurons survive for more than 4 months and express neuronal markers typical of the mature echinoderm CNS. CONCLUSIONS: A hypothesis is formulated that CNS regeneration via activation of radial glial cells may represent a common capacity of the Deuterostomia, which is not invoked spontaneously in higher vertebrates, whose adult CNS does not retain radial glial cells. Potential implications for biomedical research aimed at finding the cure for human CNS injuries are discussed."
Neural cells and their progenitors in regenerating zebrafish spinal cord
2020
The zebrafish (Danio rerio), among all amniotes is emerging as a powerful model to study vertebrate organogenesis and regeneration. In contrast to mammals, the adult zebrafish is capable of regenerating damaged axonal tracts; it can replace neurons and glia lost after spinal cord injury (SCI) and functionally recover. In the present paper, we report ultrastructural and cell biological analyses of regeneration processes after SCI. We have focused on event specific analyses of spinal cord regeneration involving different neuronal and glial cell progenitors, such as radial glia, oligodendrocyte progenitors (OPC), and Schwann cells. While comparing the different events, we frequently refer to previous ultrastructural analyses of central nervous system (CNS) injury in higher vertebrates. Our data show (a) the cellular events following injury, such as cell death and proliferation; (b) demyelination and remyelination followed by target innervation and regeneration of synaptic junctions and c) the existence of different progenitors and their roles during regeneration. The present ultrastructural analysis corroborates the cellular basis of regeneration in the zebrafish spinal cord and confirms the presence of both neuronal and different glial progenitors.