Long-term embryonic spinal cultures were developed and analyzed at sequential times in vitro, namely after 1, 2, and 3 weeks. Spatial and temporal regulation of neuronal and non-neuronal markers was investigated by immunocytochemical and Western blotting analysis using antibodies against: a) the non-phosphorylated epitope of neurofilament H (SMI32 antibody); b) the enzyme choline acetyltransferase, to localize motoneurons and cholinergic interneurons; c) the enzyme glutamic acid decarboxylase 67, to identify GABAergic interneurons; d) human eag-related gene (HERG) K(+) channels, which appear to be involved in early stages of neuronal and muscle development; e) glial fibrillary acidic protein, to identify mature astrocytes; f) myelin basic protein, to identify the onset of myelination by oligodendrocytes. To examine the development of muscle acetylcholine receptors clusters in vitro, we incubated live cultures with tetramethylrhodamine isothiocyanate-labeled alpha-bungarotoxin, and we subsequently immunostained them with SMI32 or with anti-myosin antibodies. Our results indicate that the developmental pattern of expression of these markers in organotypic cultures shows close similarities to the one observed in vivo. Therefore, spinal organotypic cultures provide a useful in vitro model system to study several aspects of neurogenesis, gliogenesis, muscle innervation, and synaptogenesis.

Spinal circuits formation: a study of developmentally regulated markers in organotypic cultures of embryonic mouse spinal cord / Avossa, D.; Rosato-Siri, M.; Mazzarol, F.; Ballerini, L.. - In: NEUROSCIENCE. - ISSN 0306-4522. - 122:2(2003), pp. 391-405. [10.1016/j.neuroscience.2003.07.006]

Spinal circuits formation: a study of developmentally regulated markers in organotypic cultures of embryonic mouse spinal cord

Avossa, D.;Rosato-Siri, M.;Mazzarol, F.;Ballerini, L.
2003-01-01

Abstract

Long-term embryonic spinal cultures were developed and analyzed at sequential times in vitro, namely after 1, 2, and 3 weeks. Spatial and temporal regulation of neuronal and non-neuronal markers was investigated by immunocytochemical and Western blotting analysis using antibodies against: a) the non-phosphorylated epitope of neurofilament H (SMI32 antibody); b) the enzyme choline acetyltransferase, to localize motoneurons and cholinergic interneurons; c) the enzyme glutamic acid decarboxylase 67, to identify GABAergic interneurons; d) human eag-related gene (HERG) K(+) channels, which appear to be involved in early stages of neuronal and muscle development; e) glial fibrillary acidic protein, to identify mature astrocytes; f) myelin basic protein, to identify the onset of myelination by oligodendrocytes. To examine the development of muscle acetylcholine receptors clusters in vitro, we incubated live cultures with tetramethylrhodamine isothiocyanate-labeled alpha-bungarotoxin, and we subsequently immunostained them with SMI32 or with anti-myosin antibodies. Our results indicate that the developmental pattern of expression of these markers in organotypic cultures shows close similarities to the one observed in vivo. Therefore, spinal organotypic cultures provide a useful in vitro model system to study several aspects of neurogenesis, gliogenesis, muscle innervation, and synaptogenesis.
2003
122
2
391
405
Avossa, D.; Rosato-Siri, M.; Mazzarol, F.; Ballerini, L.
File in questo prodotto:
File Dimensione Formato  
2003_Neuroscience.pdf

non disponibili

Tipologia: Versione Editoriale (PDF)
Licenza: Non specificato
Dimensione 890.41 kB
Formato Adobe PDF
890.41 kB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.11767/13923
Citazioni
  • ???jsp.display-item.citation.pmc??? 19
  • Scopus 58
  • ???jsp.display-item.citation.isi??? 55
social impact