Ventral mesencephalic grafts in the neostriatum of the weaver mutant mouse: Structural molecule and receptor studies

L. C. Triarhou, C. Solà, G. Mengod, F. J. Garcia-Ladona, B. Landwehrmeyer, Bernardino Ghetti, J. M. Palacios

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Mesencephalic cell suspensions were prepared from E12 wild-type (+/+) mouse embryos and stereotaxically implanted into the dorsal neostriatum of weaver mutant mice (wv/wv), which have a genetic mesostriatal dopamine (DA) deficiency. Survival of DA neurons in the grafts was documented by tyrosine hydroxylase (TH) immunocytochemistry. Axon growth was monitored by immunocytochemistry using a battery of antibody markers, and the cellular localization of structural protein and receptor RNA transcripts was studied by in situ hybridization histochemistry using [32P]oligonucleotide probes. The cell suspension grafts exhibited strong immunoreactivity for neural cell adhesion molecule (NCAM), growth-associated phosphoprotein GAP-43, microtubule-associated protein 2 (MAP2), β-amyloid protein precursor (βAPP), and phosphorylated neurofilament epitopes (clone SMI-31); intermediate-to-high levels of immunoreactivity were seen for synaptophysin. High levels of hybridization were found in the grafts for the RNA transcripts of GAP-43, MAP2, and isoforms βAPP695, βAPP714 and βAPP751 of the βAPP. No hybridization signal was detected in the grafts for DA D2 or neurotensin receptor mRNAs, both of which are normally expressed by nigral DA neurons. DA receptor autoradiography using the D2/D3 agonist [3H]CV 205-502 as a ligand showed no binding in the transplants, indicating an apparent abnormality of grafted cells; neurotensin binding sites, labeled with [125I]neurotensin, were visualized in the suspensions, indicating the possibility that receptors could be present but that RNA message levels might be too low to allow detection. These findings offer a molecular correlate of axonal, dendritic and structural protein expression by transplanted mesencephalic neurons; further, they suggest that specific functional properties of grafted nigral cells are maintained after transplantation, while other aspects of their cellular biology may be compromised.

Original languageEnglish
Pages (from-to)39-48
Number of pages10
JournalCell Transplantation
Volume4
Issue number1
DOIs
StatePublished - 1995

Fingerprint

Neurologic Mutant Mice
Neostriatum
Grafts
Transplants
Molecules
RNA
Neurons
GAP-43 Protein
Proteins
Neurotensin
Suspensions
Microtubule-Associated Proteins
Amyloid beta-Protein Precursor
Dopaminergic Neurons
Substantia Nigra
Immunohistochemistry
Neurotensin Receptors
Cytology
Epitopes
Neural Cell Adhesion Molecules

Keywords

  • Dopamine
  • GAP-43
  • MAP2
  • Mouse
  • N-CAM
  • Neostriatum
  • Neural graft
  • Neurofilaments
  • Neurological mutant
  • Neurotensin
  • Synaptophysin
  • Weaver
  • βAPP

ASJC Scopus subject areas

  • Cell Biology
  • Biomedical Engineering
  • Transplantation

Cite this

Ventral mesencephalic grafts in the neostriatum of the weaver mutant mouse : Structural molecule and receptor studies. / Triarhou, L. C.; Solà, C.; Mengod, G.; Garcia-Ladona, F. J.; Landwehrmeyer, B.; Ghetti, Bernardino; Palacios, J. M.

In: Cell Transplantation, Vol. 4, No. 1, 1995, p. 39-48.

Research output: Contribution to journalArticle

Triarhou, L. C. ; Solà, C. ; Mengod, G. ; Garcia-Ladona, F. J. ; Landwehrmeyer, B. ; Ghetti, Bernardino ; Palacios, J. M. / Ventral mesencephalic grafts in the neostriatum of the weaver mutant mouse : Structural molecule and receptor studies. In: Cell Transplantation. 1995 ; Vol. 4, No. 1. pp. 39-48.
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AU - Ghetti, Bernardino

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N2 - Mesencephalic cell suspensions were prepared from E12 wild-type (+/+) mouse embryos and stereotaxically implanted into the dorsal neostriatum of weaver mutant mice (wv/wv), which have a genetic mesostriatal dopamine (DA) deficiency. Survival of DA neurons in the grafts was documented by tyrosine hydroxylase (TH) immunocytochemistry. Axon growth was monitored by immunocytochemistry using a battery of antibody markers, and the cellular localization of structural protein and receptor RNA transcripts was studied by in situ hybridization histochemistry using [32P]oligonucleotide probes. The cell suspension grafts exhibited strong immunoreactivity for neural cell adhesion molecule (NCAM), growth-associated phosphoprotein GAP-43, microtubule-associated protein 2 (MAP2), β-amyloid protein precursor (βAPP), and phosphorylated neurofilament epitopes (clone SMI-31); intermediate-to-high levels of immunoreactivity were seen for synaptophysin. High levels of hybridization were found in the grafts for the RNA transcripts of GAP-43, MAP2, and isoforms βAPP695, βAPP714 and βAPP751 of the βAPP. No hybridization signal was detected in the grafts for DA D2 or neurotensin receptor mRNAs, both of which are normally expressed by nigral DA neurons. DA receptor autoradiography using the D2/D3 agonist [3H]CV 205-502 as a ligand showed no binding in the transplants, indicating an apparent abnormality of grafted cells; neurotensin binding sites, labeled with [125I]neurotensin, were visualized in the suspensions, indicating the possibility that receptors could be present but that RNA message levels might be too low to allow detection. These findings offer a molecular correlate of axonal, dendritic and structural protein expression by transplanted mesencephalic neurons; further, they suggest that specific functional properties of grafted nigral cells are maintained after transplantation, while other aspects of their cellular biology may be compromised.

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