Survival and death of mature avian motoneurons in organotypic slice culture: Trophic requirements for survival and different types of degeneration

Brunet, Núria; Tarabal, Olga; Portero-Otı́n, Manel; Oppenheim, Ronald W.; Esquerda, Josep E.; Calderó, Jordi

doi:10.1002/cne.21157

Cited by 33 publications

(53 citation statements)

References 129 publications

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“…The neurotrophic agent glial cell line-derived neurotrophic factor (GDNF; Alomone Labs, Jerusalem, Israel) at a concentration of 50 ng/ml was added to the culture medium in order to promote the survival of MNs. As we have previously shown, MNs massively die in this culture system in the absence of exogenous trophic support (Brunet et al, 2007), whereas GDNF promotes the long-term survival (about 70%) and growth of MNs in organotypic cultures of E16 chick embryo spinal cord maintained for at least 28 days in vitro (DIV). Culture dishes were incubated at 378C in a 5% CO 2 -95% O 2 humidified incubator.…”

Section: Organotypic Slice Cultures Of Spinal Cordsupporting

confidence: 51%

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Development of microglia in the chick embryo spinal cord: Implications in the regulation of motoneuronal survival and death

Calderó¹,

Brunet²,

Ciutat³

et al. 2009

J of Neuroscience Research

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The role of microglia during normal development of the nervous system is still not well understood. In the present study, a chick embryo model was used to examine the development of microglia in the spinal cord and characterize their changes in response to naturally occurring and pathological death of motoneurons (MNs). The microglial response to MN axotomy and the effects of microglial activation on MN survival were also studied. We found that: 1) macrophages/microglial cells were present in the spinal cord at early developmental stages (E3) and that they were recruited after normal and induced MN apoptosis; 2) although many microglial cells were seen phagocytosing apoptotic bodies, a proportion of dying cells were devoid of engulfing microglia; 3) axotomy of mature MNs was accompanied by microglial activation in the absence of MN death; 4) excitotoxic (necrotic) MN death provoked a rapid and massive microglial recruitment with phagocytic activity; 5) lipopolysaccharide-induced microglial activation in vivo resulted in the death of immature, but not mature, microglia; and 6) overactivation of microglia modulated the survival of mature MNs, either by killing them or by enhancing their vulnerability to die in response to a mild injury. Taken together, these observations indicate that normal microglia do not play an active role in triggering apoptosis of developing MNs. Rather, they act as phagocytes for the removal of dying cells during the process of programmed cell death.

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Section: Organotypic Slice Cultures Of Spinal Cordsupporting

confidence: 51%

“…Organotypic slice cultures were prepared from lumbar spinal cord of chick embryos on E16, as described previously (Brunet et al, 2007). Spinal cords were dissected, removed, and transversely sectioned at 350 lm with a McIlwain tissue chopper (Mickle Laboratory Engineering, Gomshall; Surrey, UK).…”

Section: Organotypic Slice Cultures Of Spinal Cordmentioning

confidence: 99%

Development of microglia in the chick embryo spinal cord: Implications in the regulation of motoneuronal survival and death

Calderó¹,

Brunet²,

Ciutat³

et al. 2009

J of Neuroscience Research

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“…In the last 10 years, it has been increasingly recognized that there are diverse mechanisms that can mediate developmental as well as injury-induced PCD (Sperandio et al, 2000;Boya and de la Rosa, 2005;Chipuk and Green, 2005;Bredesen et al, 2006;Blomgren et al, 2007;Brunet et al, 2007). In the present study, we present evidence consistent with this contemporary view by demonstrating that in the absence of a key player in type 1 apoptotic PCD (Apaf-1), developing postmitotic neurons nonetheless undergo PCD but by a nonapoptotic caspase-independent pathway that exhibits signs of autophagy.…”

Section: Discussionsupporting

confidence: 77%

Developing Postmitotic Mammalian NeuronsIn VivoLacking Apaf-1 Undergo Programmed Cell Death by a Caspase-Independent, Nonapoptotic Pathway Involving Autophagy

Oppenheim¹,

Blomgren²,

Ethell³

et al. 2008

J. Neurosci.

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Previous studies have shown that caspases and Apaf-1 are required for the normal programmed cell death (PCD) in vivo of immature postmitotic neurons and mitotically active neuronal precursor cells. In contrast, caspase activity is not necessary for the normal PCD of more mature postmitotic neurons that are establishing synaptic connections. Although normally these cells use caspases for PCD, in the absence of caspase activity these neurons undergo a distinct nonapoptotic type of degeneration. We examined the survival of these more mature postmitotic neuronal populations in mice in which Apaf-1 has been genetically deleted and find that they exhibit quantitatively normal PCD of developing postmitotic neurons. We next characterized the morphological mode of PCD in these mice and show that the neurons degenerate by a caspase-independent, nonapoptotic pathway that involves autophagy. However, autophagy does not appear to be involved in the normal PCD of postmitotic neurons in which caspases and Apaf-1 are present and functional because quantitatively normal neuronal PCD occurred in the absence of a key gene required for autophagy (ATG7). Finally, we examined the possible role of another caspase-independent type of neuronal PCD involving the apoptosis-inducing factor (AIF). Mice deficient in AIF also exhibit quantitatively normal PCD of postmitotic neurons after caspase inhibition. Together, these data indicate that, when key components of the type 1 apoptotic pathway (i.e., caspases and Apaf-1) are perturbed in vivo, developing postmitotic neurons nonetheless undergo quantitatively normal PCD by a caspase-independent pathway involving autophagy and not requiring AIF.

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“…For example, neurotrophic factors are critical for the survival of spinal motor neurons [34][35][36]. It is possible that SMN disrupts the responses of motor neurons to trophic factors, which needs to be further investigated.…”

Section: Discussionmentioning

confidence: 99%

Recapitulation of spinal motor neuron-specific disease phenotypes in a human cell model of spinal muscular atrophy

Wang

Zhang

2012

Cell Res

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Establishing human cell models of spinal muscular atrophy (SMA) to mimic motor neuron-specific phenotypes holds the key to understanding the pathogenesis of this devastating disease. Here, we developed a closely representative cell model of SMA by knocking down the disease-determining gene, survival motor neuron (SMN), in human embryonic stem cells (hESCs). Our study with this cell model demonstrated that knocking down of SMN does not interfere with neural induction or the initial specification of spinal motor neurons. Notably, the axonal outgrowth of spinal motor neurons was significantly impaired and these disease-mimicking neurons subsequently degenerated. Furthermore, these disease phenotypes were caused by SMN-full length (SMN-FL) but not SMN-∆7 (lacking exon 7) knockdown, and were specific to spinal motor neurons. Restoring the expression of SMN-FL completely ameliorated all of the disease phenotypes, including specific axonal defects and motor neuron loss. Finally, knockdown of SMN-FL led to excessive mitochondrial oxidative stress in human motor neuron progenitors. The involvement of oxidative stress in the degeneration of spinal motor neurons in the SMA cell model was further confirmed by the administration of N-acetylcysteine, a potent antioxidant, which prevented disease-related apoptosis and subsequent motor neuron death. Thus, we report here the successful establishment of an hESC-based SMA model, which exhibits disease gene isoform specificity, cell type specificity, and phenotype reversibility. Our model provides a unique paradigm for studying how motor neurons specifically degenerate and highlights the potential importance of antioxidants for the treatment of SMA.

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Survival and death of mature avian motoneurons in organotypic slice culture: Trophic requirements for survival and different types of degeneration

Cited by 33 publications

References 129 publications

Development of microglia in the chick embryo spinal cord: Implications in the regulation of motoneuronal survival and death

Development of microglia in the chick embryo spinal cord: Implications in the regulation of motoneuronal survival and death

Developing Postmitotic Mammalian NeuronsIn VivoLacking Apaf-1 Undergo Programmed Cell Death by a Caspase-Independent, Nonapoptotic Pathway Involving Autophagy

Recapitulation of spinal motor neuron-specific disease phenotypes in a human cell model of spinal muscular atrophy

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