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Differential effects of Bcl-2 overexpression on fibre outgrowth and survival of embryonic dopaminergic neurons in intracerebral transplants

Schierle, G S LU ; Leist, M; Martinou, J C; Widner, H LU ; Nicotera, P and Brundin, P LU (1999) In European Journal of Neuroscience 11(9). p.81-3073
Abstract

The causes of death of transplanted neurons are not known in detail, but apoptotic mechanisms involving caspase activation are likely to play a role. We examined whether overexpression of the anti-apoptotic protein Bcl-2 may enhance the survival of dopaminergic [tyrosine hydroxylase (TH)-immunoreactive] grafted neurons. For this purpose, we prepared cells from embryonic day 13 ventral mesencephalon (VM) of mice overexpressing human Bcl-2, or from their wild-type littermates. The bcl-2 transgene was strongly expressed in these cells, and resulted in protection of neuronal cultures from death triggered by serum deprivation or exposure to staurosporine. To model pretransplantation stress more closely in vitro, we stored dissociated... (More)

The causes of death of transplanted neurons are not known in detail, but apoptotic mechanisms involving caspase activation are likely to play a role. We examined whether overexpression of the anti-apoptotic protein Bcl-2 may enhance the survival of dopaminergic [tyrosine hydroxylase (TH)-immunoreactive] grafted neurons. For this purpose, we prepared cells from embryonic day 13 ventral mesencephalon (VM) of mice overexpressing human Bcl-2, or from their wild-type littermates. The bcl-2 transgene was strongly expressed in these cells, and resulted in protection of neuronal cultures from death triggered by serum deprivation or exposure to staurosporine. To model pretransplantation stress more closely in vitro, we stored dissociated embryonic mesencephalic cells for 8 h in the same type of medium used for intracerebral transplantation. This resulted in massive cell death as quantified by lactate dehydrogenase (LDH) release, and increased DNA fragmentation. Although this cell loss was strongly reduced by a caspase inhibitor, Bcl-2 had no significant protective effect. Finally, mesencephalic cell suspensions were xenografted into the striatum of immunosuppressed hemiparkinsonian rats. Neither the survival of TH-immunopositive transplanted neurons nor the functional recovery of the rats was improved by Bcl-2, although the Bcl-2 protein was strongly expressed in transgenic grafts 5 weeks after implantation, and dopaminergic fibre outgrowth from the grafts was significantly improved. These data suggest that cell death in neuronal transplants involves apoptotic mechanisms that can bypass negative regulation by Bcl-2.

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published
subject
keywords
Animals, Apoptosis, Brain Tissue Transplantation, Culture Media, Serum-Free, Dopamine, Enzyme Inhibitors, Female, Fetal Tissue Transplantation, Functional Laterality, Gene Expression, Genes, bcl-2, Graft Survival, Humans, Immunoblotting, Mesencephalon, Mice, Mice, Transgenic, Nerve Fibers, Neurons, Rats, Rats, Sprague-Dawley, Staurosporine
in
European Journal of Neuroscience
volume
11
issue
9
pages
9 pages
publisher
Wiley-Blackwell
external identifiers
  • scopus:0032880090
ISSN
0953-816X
DOI
10.1046/j.1460-9568.1999.00727.x
language
English
LU publication?
yes
id
bc222acb-0d46-47bc-8edb-4ffaadff4376
date added to LUP
2017-04-19 18:22:32
date last changed
2017-04-28 12:08:12
@article{bc222acb-0d46-47bc-8edb-4ffaadff4376,
  abstract     = {<p>The causes of death of transplanted neurons are not known in detail, but apoptotic mechanisms involving caspase activation are likely to play a role. We examined whether overexpression of the anti-apoptotic protein Bcl-2 may enhance the survival of dopaminergic [tyrosine hydroxylase (TH)-immunoreactive] grafted neurons. For this purpose, we prepared cells from embryonic day 13 ventral mesencephalon (VM) of mice overexpressing human Bcl-2, or from their wild-type littermates. The bcl-2 transgene was strongly expressed in these cells, and resulted in protection of neuronal cultures from death triggered by serum deprivation or exposure to staurosporine. To model pretransplantation stress more closely in vitro, we stored dissociated embryonic mesencephalic cells for 8 h in the same type of medium used for intracerebral transplantation. This resulted in massive cell death as quantified by lactate dehydrogenase (LDH) release, and increased DNA fragmentation. Although this cell loss was strongly reduced by a caspase inhibitor, Bcl-2 had no significant protective effect. Finally, mesencephalic cell suspensions were xenografted into the striatum of immunosuppressed hemiparkinsonian rats. Neither the survival of TH-immunopositive transplanted neurons nor the functional recovery of the rats was improved by Bcl-2, although the Bcl-2 protein was strongly expressed in transgenic grafts 5 weeks after implantation, and dopaminergic fibre outgrowth from the grafts was significantly improved. These data suggest that cell death in neuronal transplants involves apoptotic mechanisms that can bypass negative regulation by Bcl-2.</p>},
  author       = {Schierle, G S and Leist, M and Martinou, J C and Widner, H and Nicotera, P and Brundin, P},
  issn         = {0953-816X},
  keyword      = {Animals,Apoptosis,Brain Tissue Transplantation,Culture Media, Serum-Free,Dopamine,Enzyme Inhibitors,Female,Fetal Tissue Transplantation,Functional Laterality,Gene Expression,Genes, bcl-2,Graft Survival,Humans,Immunoblotting,Mesencephalon,Mice,Mice, Transgenic,Nerve Fibers,Neurons,Rats,Rats, Sprague-Dawley,Staurosporine},
  language     = {eng},
  number       = {9},
  pages        = {81--3073},
  publisher    = {Wiley-Blackwell},
  series       = {European Journal of Neuroscience},
  title        = {Differential effects of Bcl-2 overexpression on fibre outgrowth and survival of embryonic dopaminergic neurons in intracerebral transplants},
  url          = {http://dx.doi.org/10.1046/j.1460-9568.1999.00727.x},
  volume       = {11},
  year         = {1999},
}