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Keywords:

  • cerebellar development;
  • monoclonal antibody;
  • cerebellum;
  • Purkinje cell;
  • immunocytochemistry

Abstract

The mammalian cerebellar cortex consists of a number of parasagittal Purkinje cell compartments that can be demonstrated cytochemically. The afferent inputs to the cerebellum are also compartmentalized, and a complex but reproducible relationship exists between the afferents and the intrinsic maps. Developmental studies in the rat have shown that many of the main features of compartmentation are already established at birth, and are therefore not easily manipulated experimentally. The compartmentation antigen zebrin II is expressed selectively by Purkinje cell subsets in a range of species, including fish and primates. In this study, zebrin II immunoreactivity has been studied in the grey opossum, Monodelphis domestica, in order to develop a marsupial model of compartment formation in which the early developmental events are more readily accessible. A monoclonal antibody to zebrin II from the weakly electric fish Apteronotus recognizes a 36 kD polypeptide in homogenates of Monodelphis cerebellum that appears to be identical to the antigen in the rat.

Immunocytochemistry reveals that zebrin II in adult Monodelphis is confined exclusively to the cerebellum, where it is expressed by a subset of Purkinje cells. All regions of the cell, except the nucleus, are stained. The zebrin II+ Purkinje cells are arranged in a set of parasagittal compartments interposed by similar zebrin II compartments. In each hemicerebellum there is one zebrin II+ band abutting the midline (P1+), and two others laterally in the vermis (P2+, P3+). A fourth zebrin II+ compartment straddles the paravermian region (P4+). Three other compartments have been identified in the hemisphere (P5+, P6+, P7+). This arrangement is very similar to that found in the rat. During postnatal development, zebrin II is first expressed between P14 and P21 in Purkinje cells of the posterior lobe vermis, and spreads throughout the cerebellar cortex by P28. As in rat, there is a stage at which all Purkinje cells are zebrin II+, including those destined to be zebrin II in the adult. The mature pattern of expression emerges after P35 as immunoreactivity gradually disappears from the cells destined to become zebrin II. The adult appearance is attained only after P56. The developmental timetable is therefore similar to that in rat, but is rather more protracted. Monodelphis should prove to be a valuable experimental model in which to study the early events leading to the formation of cerebellar compartments.