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Volume 17, Number 10, Issue of May 15, 1997 pp. 3599-3609
Copyright ©1997 Society for NeuroscienceRegulation of Purkinje Cell Alignment by Reelin as Revealed with CR-50 Antibody
Received Dec. 14, 1996; revised Feb. 10, 1997; accepted Feb. 24, 1997.
Takaki Miyata1, 2, 3, Kazunori Nakajima1, Katsuhiko Mikoshiba1, 3, , and Masaharu Ogawa2, 1 Molecular Neurobiology Laboratory, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Tsukuba, Ibaraki 305, Japan, 2 Department of Physiology, Kochi Medical School, Nankoku, Kochi 783, Japan, and 3 Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108, Japan
Cerebellar Purkinje cells are generated in the ventricular zone, migrate outward, and finally form a monolayer in the cortex. In reeler mice, however, most Purkinje cells cluster abnormally in subcortical areas. Reelin, the candidate reeler gene product recognized by the CR-50 monoclonal antibody, is concentrated in a cortical zone along which Purkinje cells are aligned linearly, implying that it may regulate their alignment. We used an in vitro system and a transplantation approach to analyze the function of Reelin.
Explant culture for 7 d of cerebella isolated from wild-type and reeler mice at embryonic day 13 (E13) reproduced in a phenotype-dependent manner the two distinct arrangement patterns (linear vs clustered) of Purkinje cells. Extensive CR-50 binding to wild-type explants converted the linear pattern into a reeler-like, clustered pattern. On the other hand, when reeler explants lacking Reelin were crowned with an artificial layer of Reelin+ granule cells, some Reelin molecules were distributed into a superficial zone of the reeler explants, and Purkinje cells formed a linear pattern along the Reelin-rich overlay. This "rescue" effect was also inhibited by CR-50. Hence, Reelin is involved in the Purkinje cell alignment, and the lack of this activity may explain the malformation in reeler cerebella.
We further injected Reelin+ granule cells into the fourth ventricle of E12-13 mice. Extensive incorporation of the injected Reelin+ cells into the ventricular zone, but not of Reelin
cells, forced Purkinje cells of the host cerebella to form an aberrant layer, suggesting that premigratory Purkinje cells may already be responsive to Reelin or Reelin-related signals.
Key words: cerebellum; layer formation; cell migration; reeler mutant mouse; reelin; CR-50; Purkinje cell; granule cell; explant culture; transplantation
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