Purkinje cells monoclonal antibodie

Fig. 23. Section of reeler mutant mouse cerebellum stained with monoclonal antibody 4C11 against the P400 protein. Note stained Purkinje cells in the cortex (CX) and in the central mass of dislocated cells (DP). Bar = 200 jum. Maeda et al. (1989).

Several other markers are only present in zonally distributed subpopulations of Purkinje cells (see also Section 6.1.3.)- The monoclonal antibody B1 of Ingram et al. 

Levine et al. (1986), who used another monoclonal antibody against GD3, found immunoreactivity of reactive astrocytes in mouse mutants, but failed to observe a reaction within the Purkinje cells. These different results probably are due to differences in fixation (Reynolds and Wilkin, 1988). 

P75 nerve growth factor-receptor protein (NGF-R) is present in developing and adult Purkinje cells. Yan and Johnson (1988) and Cohen-Cory et ah (1989) described and reviewed the development of NGF-R in rat cerebellum. Low affinity NGF-R immunoreactivity has been demonstrated with species-specific monoclonal antibodies in Purkinje cells of adult rats (Pioro and Cuello, 1988, 1990 Pioro et ah, 1991 Fusco et ah, 1991 Dusart et ah, 1994), monkey and human brain (Mufson et ah, 1991). Immunoreactivity was present in the somata, dendrites and the proximal axon of the Purkinje cells. Additional immunoreactivity in granule cells was reported by Vega et ah... 

Fig. 66. A. Monoclonal antibody Ral-303 recognizes neurons in the granule cell layer (g), but not in the Purkinje cell (p) or molecular (m) layers of the rat cerebellar cortex. Scale bar = 100 //m. B. The morphology of Rat-303-positive neurons matches that described for Golgi II cells a large cell body emitting relatively stout dendrites from many points over the cell circumference (13). Scale bar = 10 im. Hockfield (1987).

Fig. 69. A. Monoclonal antibody Rat-302 recognizes a subset of neurons, later identified as unipolar brush cells, restricted to the granular layer of the flocculus and the vermis of rat cerebellum (arrows), whereas in other areas of the cerebellum no positive cells are found. B. In contrast, antibody Rat-303 recognizes Golgi-II cells in the granular layer (g) in the entire cerebellum. C. Rat-302 also recognizes Purkinje cells outside the caudal vermis and the flocculus. D. Rat-302 positive cells in the vermis. E and F. Unipolar brush cells recognized by Rat-302 have a round cell body and short dendrites ending in a spray of appendages (arrows), g, granular layer m, molecular layer p, Purkinje cell layer. Scale bars 500 fim in A and B, 50 /rm in C and D. 10 /rm in E and F. Hockfield (1987).

Fig. 85. A. Drawing displaying the distribution of mossy rosettes (dots) immunoreactive to monoclonal choline-acetyltransferase (ChAT) antibody. The section (40 fim thick) was cut sagittally through the middle vermis of rat cerebellum. A considerable number of immunoreactive mossy terminals are observed in lobules I through IXab, although they are much fewer than in lobules IXc and X. Calibration bar = 1 mm. B. Drawing of part of lobule IXab shows the overall distribution of immunoreactive fibers. Arrows indicate mossy fibers with glomerular rosettes. Small and large arrowheads point to some varicose fibers distributing in or near the Purkinje cell layer (PCL) and in the molecular layer (ML), respectively. The ML fibers are most frequently observed in this lobule and tend to be restricted to the inner half of the layer. Calibration bar = 200 jum. Ojima et al. (1989).

The distribution of 5 -nucleotidase (5 -N) (Section 3.5.) in alternate longitudinal bands of high and low enzyme activity in the molecular layer of the cerebellar cortex of the mouse (Scott, 1963,1964,1965,1967) was the first evidence for the biochemical compart-mentalization of the cerebellar cortex. The pattern of 5 -N-positive and -negative zones is complete in the sense that it is present in all the lobules of vermis and hemisphere and unequivocal, because, in the mouse at least, the bands are clearly delineated (Marani, 1986). The 5 -N band pattern is very similar, if not identical, to the more recently described distribution of Purkinje cells in the rat, reacting with Purkinje cell-specific monoclonal antibodies to Zebrin-I (mabQl 13) (Eisenman and Hawkes, 1989). 

The epitopes recognized by Hawkes family of monoclonal antibodies known as the anti-Zebrins are localized on Purkinje cells (see Section 3.1.8.). Zonal patterns that are identical or very similar to Zebrin I and II have been described for the distribution of 5 -nucleotidase (see above), the p75 low affinity nerve growth factor receptor protein in the rat (Section 3.1.10., Fig. 38), protein kinase C delta (Fig. 133) (see Section 3.1.5.) and the B30 antibody of Stainier and Gilbert (1989) (see Section 3.1.8.). Immunoreactiv-ity in mouse Purkinje cells for an antibody against HNK is partially congruent with the Zebrin negative Purkinje cells, but Zebrin+/HNK-l- Purkinje cells also exist (Hawkes, 1992 Eisenman and Hawkes, 1993). The similarity between the Zebrin pattern and the transient zonal patterns in the development of the Purkinje cell specific marker L7 is discussed in Section 6.2. 

Bands of P-path immunoreactive Purkinje cells alternate with zebrin II immuno-reactive neurons in the cerebellum of the mouse (Leclerc et al., 1992) (Fig. 134). In the P3+ band in the anterior vermis, lobule VII, VIII and dorsal IX, the P4+ band in the lobules V and VIII and the P2+ band in dorsal lobule IX the two epitopes are colocalized. The B1 monoclonal antibody of Ingram et al. (1985) also detects a subset of Purkinje cells in monkey cerebellum, but their distribution did not correspond to the distribution of Zebrin I in the squirrel monkey (Leclerc et al., 1990) or to AChE as reported by Hess and Voogd (1986). 

Hawkes R, Leclerc N (1987) Antigenic map of the rat cerebellar cortex the distribution of sagittal bands as revealed by monoclonal anti-Purkinje cell antibody mabQl 13. J. Comp. Neurol, 256, 29-41. 

Langley OK, Sternberger NH, Sternberger LA (1988) Expression of neurofilament proteins by Purkinje cells ultra-structural immunolocalization with monoclonal antibodies. Brain Res., 457, 12-20. 

Matsushita M, Ragnarson B, Grant G (1991) Topographic relationship between sagittal Purkinje cell bands revealed by a monoclonal antibody to zebrin I and spinocerebellar projections arising from the central cervical nucleus in the rat. Exp. Brain Res., 84, 133-141. 

Trisphosphate (InsPj) receptor in mouse cerebellar Purkinje cells using three monoclonal antibodies. Cell Structure, Function, 13, 163-173. 

Royds JA, Ironside JW, Warnaar SO, Taylor CB, Timperley WR (1987) Monoclonal antibody to aldolase C A selective marker for Purkinje cells in the human cerebellum. Neuropath. Appl Neurobiol, 13, 11-21. 

Weber A, Schachner M (1982) Development and expression of cytoplasmic antigenes in Purkinje cells recognized by monoclonal antibodies. Cell Tissue Res., 227, 659-676. 

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