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  • 1
    Electronic Resource
    Electronic Resource
    Development and fine structure of murine Purkinje cells in dissociated cerebellar cultures: dendritic differentiation, synaptic maturation, and formation of cell-class specific features (1997)
    Springer
    Anatomy and embryology 197 (1997), S. 31-50 
    Details
    ISSN: 1432-0568
    Keywords: Key words Purkinje neuron ; Synaptogenesis ; Dendritogenesis ; L7 ; Cerebellum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The morphological differentiation of E16 murine Purkinje cells (PCs) in dissociated cerebellar cultures was analyzed by light and electron microscopic immunocytochemistry after 2–5 weeks in vitro (wiv), with particular emphasis on dendritic differentiation, synaptic maturation, and formation of stereotypical fine structural features. This study complements a companion paper on the features of PCs after 1 wiv. After 2 wiv, the PCs have an eccentric nucleus and the cytoplasmic organelles appear immature; the axon has a distinct initial segment and beaded axon collaterals but its boutons still contain sparse synaptic vesicles; dendrites show few bifurcations and tufts of spiny branchlets. After 3 wiv, the PCs display a centered nucleus, an extensive hypolemmal cisternal system, and stacks of up to four cisterns of granular endoplasmic reticulum; there is an increased number of dendritic bifurcations, spiny branchlets, mature spines, and axonal branches; dendritic tips still contain vesicle clusters, suggesting growth, and many synapses and afferent boutons continue to display immature features. After 4 wiv, elaborate perinucleolar coiled body rosettes, subsurface cistern-mitochondrion complexes and large stacks of granular endoplasmic reticulum finally appear within the soma; dendrites show a further increase in the numbers of bifurcations, segments and spines; most spines are synaptic and show mature features; afferent synapses are differentially distributed; PC boutons consistently display mature features and show a considerable degree of target specificity, although naked spines and reduced glial sheaths persist. After 5 wiv, PCs do not show further maturation and some dystrophic features appear. We conclude that under standard conditions and despite the disruption of normal tissue organization, PCs in dissociated cultures differentiate maximally after 4 wiv, at which stage they display many of the light and electron microscopic features that characterize mature PCs in situ. This prolonged developmental time-frame resembles that in the normal cerebellum. In view of the increasing usage of dissociated cerebellar cultures to study aspects of neuronal differentiation, synaptic activation and neuronal-glial interactions, an elucidation of the neurocytology of dissociated cerebellar cultures as presented in this study provides important clues for the interpretation of experimental data.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004290050118
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  • 2
    Electronic Resource
    Electronic Resource
    Development and fine structure of murine Purkinje cells in dissociated cerebellar cultures: neuronal polarity (1997)
    Springer
    Anatomy and embryology 197 (1997), S. 9-29 
    Details
    ISSN: 1432-0568
    Keywords: Key words Cerebellum ; Dendritogenesis ; Filopodia ; Growth cone ; L7 ; Polarization ; Purkinje neuron ; Synaptogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Cerebellar Purkinje cells (PC) display a highly distinctive form of polarity. We have cultured murine PCs from dissociated E16 cerebellar anlagen for 1 week to investigate the early stages of neuronal compartmentalization and synaptic interactions, features which are important for the establishment of neuronal polarity. To unequivocally identify the PCs we utilized light and electron microscopic immunocytochemistry with an antiserum to the cell class-specific marker L7/pcp2 gene product. The PCs typically show a single, long axon, numerous short appendages classified as filopodia and protospines, and a small number of protodendrites. The nucleus is positioned asymmetrically in both the horizontal and vertical axes of the soma. The Golgi apparatus, coated and uncoated vesicles, and mitochondria are prominent ultrastructural features, while the endoplasmic reticulum is highly fragmented. The cell body receives rudimentary synapses on its smooth surfaces and appendages and no consistent morphological differences were detected between these elementary contacts. The axon is clearly identifiable; it emanates from either the cell body or a protodendrite, bifurcates at predominantly right angles, forms beaded collaterals, and terminates with relatively large growth cones. The varicosities of the PC axon contain pleomorphic synaptic vesicles and form rudimentary synapses primarily with the dendritic shafts of immunonegative neurons. The protodendrites are short, quickly tapering and sparsely branched; they emit numerous filopodia and immature spines and terminate with small growth cones. Rudimentary synapses are received on the proximal dendritic shafts and filopodia, and more mature synapses occur frequently on protospines. With few exceptions, PCs lie atop an astrocytic bed layer and glial processes are apposed to the various aspects of the PC body left free by the afferent axons. By contrast, PC processes are largely free of glial sheaths. We conclude that the ”stellate stage” of PC development in situ is replicated rather faithfully in culture and that PCs have established polarity and have begun to form intercellular contacts by 1 week in vitro. Moreover, the PCs are already morphologically distinct from other cell types in the 1-week cultures, although they have yet to develop the differentiated features that distinguish mature PCs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004290050117
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    Paper (German National Licenses)
    Fulltext
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