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Cerebellar Purkinje cells from the Lurcher mutant and wild‐type mouse grown in vitro: A light and electron microscope study
Author(s) -
Doughty Martin L.,
Patterson Lilian,
Caddy Keith W. T.
Publication year - 1995
Publication title -
journal of comparative neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.855
H-Index - 209
eISSN - 1096-9861
pISSN - 0021-9967
DOI - 10.1002/cne.903570114
Subject(s) - biology , electron microscope , cerebellum , neuroscience , mutant , in vitro , cerebellar cortex , microbiology and biotechnology , genetics , optics , gene , physics
Lurcher is an autosomal semidominant murine mutation. Lurcher heterozygotes, (+/Lc) lose all their cerebellar Purkinje cells by adulthood. Explants from 2 days postnatal (P2) wild‐type (+/+) and +/Lc cerebellar cortex were grown in vitro to investigate the role of local neuronal environment and afferent input on the degenerating +/Lc Purkinje cell. In Lurcher explants, Purkinje cells were maintained for up to 25 days in vitro. No significant difference was observed between +/+ and +/Lc Purkinje cell numbers from 10 to 20 days in vitro, as revealed by calbindin‐D immunoreactivity. Growing +/Lc explants in association with + / + explants resulted in no significant difference in Purkinje cell survival (10–20 days in vitro). Image analysis of the gross morphology of calbindin‐D‐immunostained Purkinje cells from +/+ and +/Lc explants grown in vitro revealed a significant decrease in the total area and dendritic lengths of +/Lc Purkinje cells (15 and 20 days in vitro). The fine structure of +/Lc and +/+ Purkinje cells was examined under the electron microscope (10–25 days in vitro). No difference in ultrastructure was observed between +/Lc and +/+ Purkinje cells grown in vitro, and many features similar to normal Purkinje cell development in vivo were present. These included monosynaptic parallel fibre synapses with Purkinje cell dendritic spines, other interneuron synapses with Purkinje cell dendrites and soma, astroglial investment, and minimal extracellular space in the neuropil. Unusual features observed included a persistence of the perisomatic spines in some Purkinje cells, an absence of Nissl bodies in the Purkinje cell perikaryon, naked Purkinje cell dendritic spines, and occasional heterol, ogous synapses. The results are discussed in the light of previous chimeric analysis of the Lurcher mutation, and a hypothesis is put forward to explain the survival of + /Lc Purkinje cells in vitro. © 1995 Wiley‐Liss, Inc.