Structure and locomotion of adult in vitro regenerated spiral ganglion growth cones – A study using video microscopy and SEM

Anderson, Malin; Boström, Marja; Pfaller, Kristian; Glueckert, Rudolf; Schrott‐Fischer, Anneliese; Gerdin, Bengt; Rask‐Andersen, Helge

doi:10.1016/j.heares.2006.03.014

Cited by 18 publications

(20 citation statements)

References 44 publications

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“…By comparison, the yield after dissociation was 10% for adult rats, and 1% to 6% of the seeded neurons survived (Lefebvre et al, 1991); 20-30 neurons per spiral ganglion survived for adult guinea pigs (Anderson et al, 2006). On an absolute scale, all these yields are low, as may be expected for adult neurons (Banker and Goslin, 1998).…”

Section: Discussionmentioning

confidence: 89%

“…The pioneering work on culturing dissociated spiral ganglion neurons was carried out more than a decade ago with embryonic rats and chicken (Lefebvre et al, 1990a;Yamaguchi and Ohmori, 1990). Various protocols have since been established for culturing spiral ganglion neurons from a range of species and ages, such as embryonic mice (Rabejac et al, 1994;Vazquez et al, 1994); neonatal mice (Kita et al, 2005;Lin et al, 1998;Mo and Davis, 1997;Whitlon et al, 2006), rats (Dazert et al, 1998;Hegarty et al, 1997;Lefebvre et al, 1990b;Malgrange et al, 1996;Marzella et al, 1997;Ripoll and Rebillard, 1997;Rome et al, 1999;Zheng et al, 1995), and gerbils (Lin, 1997); and adult rats (Lefebvre et al, 1991), guinea pigs (Anderson et al, 2006;Rask-Andersen et al, 2005), and humans (Rask-Andersen et al, 2005). Sensory epithelia have also been cultured from the cochlea of adult guinea pigs (Zhao, 2001); most experiments with spiral ganglion neurons, however, have been conducted with neonatal samples, possibly because adult neurons are in general more difficult to culture (Banker and Goslin, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…In contrast, almost all protocols cited above use physiological saline for the isolation of spiral ganglion neurons and generic chemically-defined media, such as Dulbecco modified Eagle medium, together with the traditional N1 or N2 supplements (Bottenstein and Sato, 1985) for their culture. Only a single study has described culturing spiral ganglion neurons in Neurobasal medium with B27 supplement (Anderson et al, 2006), but the quantitation of survival and neurite outgrowth was not the focus of that work.…”

Section: Introductionmentioning

confidence: 99%

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Survival and stimulation of neurite outgrowth in a serum-free culture of spiral ganglion neurons from adult mice

et al. 2007

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We have developed a reliable protocol for the serum-free dissociation and culture of spiral ganglion neurons from adult mice, an important animal model for patients with post-lingual hearing loss. Pilot experiments indicated that the viability of spiral ganglion cells in vitro depended critically on the use of Hibernate medium with B27 supplement.With an optimized protocol, we obtained 2 · 10 3 neurons immediately after dissociation, or about one-fifth of those present in the intact spiral ganglion. After four days in culture, 4% of the seeded neurons survived without any exogenous growth factors other than insulin. This yield was highly reproducible in five independent experiments and enabled us to measure systematically the numbers and lengths of the regenerating neurites.Furthermore, the survival rate compared well to the few published protocols for culturing adult spiral ganglion neurons from other species. Enhanced survival and neurite outgrowth upon the addition of brain-derived neurotrophic factor and leukemia inhibitory factor demonstrated that both are potent stimulants for damaged spiral ganglion neurons in adults. This responsiveness to exogenous growth factors suggested that our culture protocol will facilitate the screening of molecular compounds as potential treatments for sensorineural hearing loss.3

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Section: Discussionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Survival and stimulation of neurite outgrowth in a serum-free culture of spiral ganglion neurons from adult mice

et al. 2007

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“…Neurotrophins, including brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3) alone [Zheng et al, 1995;Malgrange et al, 1996;Mou et al, 1997;Hegarty et al, 1997;Miller et al, 1997;Hansen et al, 2001a, b;Anderson et al, 2006;Vieira et al, 2007;Wei et al, 2007], and together with transforming growth factor ␤ and leukemia inhibitory factor [Marzella et al, 1999] may promote survival and growth of SG cells. Recently, in vivo studies have demonstrated that BDNF and fibroblast growth factor 1 promote auditory nerve survival and regrowth of peripheral processes following deafness [Miller et al, 2007;Glueckert et al, 2008].…”

Section: Introductionmentioning

confidence: 99%

Effects of Neurotrophic Factors on Growth and Glial Cell Alignment of Cultured Adult Spiral Ganglion Cells

Boström¹,

Khalifa²,

Boström³

et al. 2009

Audiol Neurotol

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Adult spiral ganglion cells were cultured in chorus to assess the influence of the neurotrophins brain-derived neurotrophic factor, neurotrophin 3 and glial cell line-derived neurotrophic factor (GDNF) on neurite growth and Schwann cell alignment. Over 1500 measurements were collected using each factor at 10 ng/ml and all three in combination. Evaluation was made with GDNF at concentrations of up to 100 ng/ml. Neurite dimensions were assessed at days 5, 7, 9 and 11 using a computer-based program (Axon Analyzer). GDNF had a strong effect on spiral ganglion cell growth almost attaining the level of all three factors in combination. GDNF increased glial cell alignment and nerve bundle formation. Results show the potential of GDNF to maintain and possibly restore auditory nerve integrity.

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“…Our data support this link and extend similar studies in native sensory systems governed by neurogenins. In addition to the effects in retinal ganglion cells discussed above, DCC-mediated chemoattraction plays a vital role in interneurons of the dorsal column [37], vagal sensory neurons in the gut [38,39], and primary auditory neurons in the cochlea [17,40]. However, the expression of Neurog1 and DCC alone is not predictive of netrin chemoattraction.…”

mentioning

confidence: 99%

Netrin-1-Mediated Axon Guidance in Mouse Embryonic Stem Cells Overexpressing Neurogenin-1

Hill

Purcell

Liu

et al. 2012

Stem Cells and Development

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Stem cell therapy holds great promise for treating neurodegenerative disease, but major barriers to effective therapeutic strategies remain. A complete understanding of the derived phenotype is required for predicting cell response once introduced into the host tissue. We sought to identify major axonal guidance cues present in neurons derived from the transient overexpression of neurogenin-1 (Neurog1) in mouse embryonic stem cells (ESCs). Neurog1 upregulated the netrin-1 axon guidance receptors DCC (deleted in colorectal cancer) and neogenin (NEO1). Quantitative polymerase chain reaction results showed a 2-fold increase in NEO1 mRNA and a 36-fold increase in DCC mRNA in Neurog1-induced compared with control ESCs. Immunohistochemistry indicated that DCC was primarily expressed on cells positive for the neuronal marker TUJ1. DCC was preferentially localized to the cell soma and growth-cones of induced neurons. In contrast, NEO1 expression showed less specificity, labeling both TUJ1-positive and TUJ1-negative cells as well as uninduced control cells. Axonal outgrowth was directed preferentially toward aggregates of HEK293 cells secreting a recombinant active fragment of netrin-1. These data indicate that DCC and NEO1 are downstream products of Neurog1 and may guide the integration of Neurog1-induced ESCs with target cells secreting netrin-1. Differential expression profiles for netrin receptors could indicate different roles for this guidance cue on neuronal and non-neuronal cells.

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Structure and locomotion of adult in vitro regenerated spiral ganglion growth cones – A study using video microscopy and SEM

Cited by 18 publications

References 44 publications

Survival and stimulation of neurite outgrowth in a serum-free culture of spiral ganglion neurons from adult mice

Survival and stimulation of neurite outgrowth in a serum-free culture of spiral ganglion neurons from adult mice

Effects of Neurotrophic Factors on Growth and Glial Cell Alignment of Cultured Adult Spiral Ganglion Cells

Netrin-1-Mediated Axon Guidance in Mouse Embryonic Stem Cells Overexpressing Neurogenin-1

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