Antigenic compartmentation in the mouse cerebellar cortex: Zebrin and HNK‐1 reveal a complex, overlapping molecular topography

Abstract

Two monoclonal antibodies--anti-zebrin I and anti-HNK-1--have been used to study the compartmentation of the mouse cerebellar cortex. As in other species, the pattern of localization of the Purkinje cell specific antigen zebrin I is confined to a subset of Purkinje cells that are organized into parasagittal bands. The basic pattern consists of two abutting paramedian bands (P1+) and up to three additional vermal bands on either side (P2(+)-P4+). This pattern is altered in the vermal regions of lobules X and VI-VII where all Purkinje cells are immunoreactive. In the hemisphere there are three additional bands present (P5(+)-P7+) plus two shorter bands in the paravermal area (P4b+ and P5a+) that extend from the paramedian lobule through the lobulus simplex. This pattern is very similar, but perhaps not identical, to that previously described for the rat. These results suggest a common mammalian plan for the expression and localization of zebrin I. By using a monoclonal antibody to an epitope associated with HNK-1, we have now identified a novel pattern of compartmentation in mouse cerebellum. The HNK-1 epitope is expressed most notably on Purkinje cells and Golgi cells. The molecular layer immunoreactivity associated with the Purkinje cell dendrites varies in intensity in a systematic and reproducible fashion. This reveals a novel cerebellar compartmentation that is sometimes complementary, sometimes overlapping, to that revealed by anti-zebrin. As a result, it is now possible to subdivide the cerebellar cortex into a still finer mosaic of antigenic patches and bands than was possible by using zebrins alone.

Golgi Cell Dendrites Are Restricted by Purkinje Cell Stripe Boundaries in the Adult Mouse Cerebellar Cortex

“…2); n ϭ 8 animals]: 4164 (97.3%) were confined to a single compartment, and 117 (2.7%) appeared to cross into the neighboring compartment (with some appearing to straddle the boundary and others with their dendritic arbors preferentially in the neighboring stripe). Restriction of Purkinje cell dendrites is not confined to zebrin II ϩ/Ϫ boundaries: a similar pattern of Golgi cell restriction was also found both at boundaries revealed by HNK-1 expression in ventral lobule IX and lobule X [the nodular zone (Eisenman and Hawkes, 1993;Marzban et al, 2004)], where all Purkinje cells are zebrin II ϩ (Fig. 3E), and at boundaries of expression of PLC␤4 (see Fig.…”

“…Golgi-Cox staining was performed according to Glaser and Van der Loos (1981) as modified by Gibb and Kolb (1998). Subsequently, the cerebella were transferred to 30% sucrose in distilled water for Ͼ48 h, sectioned at 200 -400 m on a Vibratome, and immunoperoxidase stained for zebrin II, with diaminobenzidine as the substrate (Eisenman and Hawkes, 1993). Photomicrographs were captured with a SPOT Cooled Color digital camera (Diagnostic Instruments, Sterling Heights, MI) and assembled in Adobe (San Jose, CA) Photoshop.…”

Zebrin-Immunopositive and -Immunonegative Stripe Pairs Represent Functional Units in the Pigeon Vestibulocerebellum

“…We used the nomenclature from Pakan et al (2007), whereby the most medial positive stripe is designated as P1ϩ and the number increases as the stripes move laterally to P7ϩ (Fig. 1 B) (Brochu et al, 1990;Eisenman and Hawkes, 1993;Ozol et al, 1999;Sillitoe and Hawkes, 2002) (for review, see Sillitoe et al, 2005). The width of individual stripes can vary both between animals as well as along the rostrocaudal axis of the cerebellum within animals.…”

Whole-mount Immunohistochemistry: A High-throughput Screen for Patterning Defects in the Mouse Cerebellum

“…More lateral still, a broader, weakly immunoreactive P3 ϩ is seen. [Zebrin-immunoreactive stripes are numbered from P1 ϩ medially to P7 ϩ laterally: zebrin-negative (P Ϫ ) stripes are numbered according to the immediately medial P ϩ stripe (Eisenman and Hawkes 1993). ]…”