Regional Variation in Expression of Calbindin and Inositol 1,4,5‐trisphosphate Receptor Type 1 mRNAs in the Cerebellum of the Staggerer Mutant Mouse

Abstract

The Purkinje cells in the staggerer mutant mouse have various cellular abnormalities, including reduced cell number, ectopia, smaller size and absence of dendritic spines. It is also know that some of these abnormalities exhibit regional variations in the cerebellum. In this paper we have investigated expression in the staggerer Purkinje cells of the calbindin and inositol 1,4, 5-trisphosphate receptor type 1 mRNAs by in situ hybridization. Although the transcription levels of both mRNAs were significantly reduced compared with the wild-type cells, the reduction among the Purkinje cell populations was not even, varying greatly from region to region. Purkinje cells with different transcription levels were distributed in discrete regions and arranged alternately in the mediolateral direction. Moreover, the cell bodies with higher transcription levels were larger in size and aligned in a monolayer between the granular and molecular layers, whereas those with lower levels were smaller in size, fewer in number and dispersed throughout the granular layer. These findings suggest that there is a distinct mediolateral heterogeneity in the staggerer cerebellum with respect to transcription levels of these Purkinje cell-specific molecules, which might correlate with some cytological phenotypes.

Cytological compartmentalization in the staggerer cerebellum, as revealed by calbindin immunohistochemistry for Purkinje cells

“…In addition to these neurocytological deficits, several lines of evidence suggest that development of neurochemical and neurophysiological properties is retarded, such as the persistence of the immature type of cell surface carbohydrates (Hatten and Messer, 1978;Trenkner, 1979), late peaking of glycosidase activity (Wille et al, 1983), delayed developmental conversion of neural cell adhesion molecules (Edelman and Chuong, 1982), and persistent N-methyl-D-aspartate (NMDA) receptor-mediated responses (Dupont et al, 1984;Nakagawa et al, 1996a). Furthermore, expressions of particular genes, which attain striking levels in adult wild-type PCs, remain at low or undetectable levels in the staggerer PCs; these include the inositol 1,4,5-trisphosphate receptor type 1 (InsP 3 R1), metabotropic glutamate receptor type 1␣, calmodulin, calbindin, and L7/pcp-2 (Mallet et al, 1975;Furuichi et al, 1989;Messer et al, 1990;Ryo et al, 1993;Hamilton et al, 1996;Nakagawa et al, 1996b). All these results suggest that the nuclear hormone receptor ROR␣ is a transcriptional regulator that exerts its profound effects in phenotypic differentiation of PCs, although its endogenous ligand has not been identified.…”

“…Recently we have found that the reduction of calbindin and InsP 3 R1 mRNA expressions does not occur evenly in the staggerer PCs, but instead, PC populations with different transcription levels compartmentalize the mature staggerer cerebellum in the mediolateral direction (Nakagawa et al, 1996b). To pursue the nature of the ''transcriptional'' compartments which emerge as the result of ROR␣ gene mutation, we developed a hightiter anti-calbindin antibody to compare the cytology of staggerer PCs in individual compartments.…”

“…In the staggerer cerebellum, it is often difficult to distinguish PCs from Golgi type II cells, because of the reduced size and ectopia of PCs (Sidman, 1962;Herrup and Mullen, 1979a;Nakagawa et al, 1996b) and of remarkable down-regulation of PC-specific or -abundant marker molecules (Furuichi et al, 1989;Nakagawa et al, 1996b). To examine the cytology of the staggerer PCs, we developed a high-titer antibody against calbindin, a cytosolic protein localized in the dendrite, cell body, and axon of PCs (Yamakuni et al, 1984).…”

Evidence against GABA Release from Glutamatergic Mossy Fiber Terminals in the Developing Hippocampus

“…We used primary antibodies raised against the following molecules (species immunized): mouse calbindin (rabbit and goat) (Nakagawa et al, 1996;Miura et al, 2006), rat and mouse vesicular glutamate transporter 1 (VGluT1; guinea pig and goat) (Miyazaki et al, 2003;Miura et al, 2006), mouse VGAT (rabbit, guinea pig, and goat) (Miyazaki et al, 2003;Miura et al, 2006), mouse GAD67/65 (Yamada et al, 2001) (rabbit), and GABA (rabbit; catalog #A2052; Sigma). For specificity control, VGAT and GAD antibodies were preabsorbed by overnight incubation with 50 -100 g/ml of antigens (Yamada et al, 2001;Miyazaki et al, 2003).…”

Cerebrospinal Fluid Calbindin D Concentration as a Biomarker of Cerebellar Disease Progression in Niemann-Pick Type C1 Disease

“…Immunostaining of Purkinje cells with antibodies to calbindin aides in quantifying Purkinje cell loss in various disease processes (de Barry and Gombos, 1989;Kurobe et al, 1992;Ferrer et al, 1993;Ishikawa et al, 1995;Maguire-Zeiss et al, 1995;Nakagawa et al, 1996;Nag and Wadhwa, 1999;Barski et al, 2003;Haworth et al, 2006;Laure-Kamionowska and Ma sli nska, 2009;Lee et al, 2010;Verdes et al, 2010;WierzbaBobrowicz et al, 2011). Indeed, decreases in calbindin immunoreactivity may be seen in disease prior to the loss of Purkinje cells and the onset of ataxia (Vig et al, 1998;Barski et al, 2003).…”