cDNA cloning and characterization of three genes uniquely expressed in cerebellum by Purkinje neurons

Abstract

The characteristics that distinguish the different neuronal cell types of an organism are believed to be primarily determined by unique patterns of cellular gene expression. The identification of cell-type specific molecules should therefore provide a good basis for understanding the biology of specific neuron types. In this paper, we describe the isolation of cDNA clones corresponding to mRNA uniquely expressed by Purkinje cells in mature mouse cerebellum. Three cDNA clones were selected from a library of normal mouse cerebellar cDNA by virtue of their failure to hybridize to mRNA sequences from the cerebella of Purkinje cell degeneration (pcd) mice. The cDNA clones were shown by in situ and Northern hybridization to correspond with mRNA present in Purkinje cells but absent or at low levels in other cell types of the cerebellum. By sequence analysis, clone PCD29 was determined to encode the calcium-binding protein calbindin-D28K. Clones PCD5 and PCD6 encode previously undescribed proteins of 99 and greater than 500 amino acids, respectively. All 3 PCD clones hybridized to mouse mRNA from sources other than cerebellum; clone PCD5 was found to have the most restricted expression, as it hybridized only to mRNA from cerebellum and eye. To define potential correlations between the PCD clones and mutations in the mouse genome known to affect Purkinje cells, clones PCD5, PCD6, and PCD29 were localized to mouse chromosomes 8, 6, and 4, respectively.

Distribution of inositol 1,4,5‐trisphosphate receptor mRNA in mouse tissues

“…However, Nordquist et al [22] reported that in testis the intensity of mRNA bands hybridizing to PCD6 clone, which is part of the IP3 receptor cDNA, appeared to be almost the same as that in kidney tissue. With regard to this discrepancy, we believe that the testis preparation of Nordquist et al [22] contained the ductus deferens (= vas deferens) which is composed of smooth muscle, while our preparation did not. In ovary sections (Fig.…”

Total Expression and Dual Gene-regulatory Mechanisms Maintained in Deletions and Duplications of the Pcdha Cluster

“…We cut 12-14 10-m thick sagittal sections at 200-m intervals from the midline, and performed in situ hybridization with probes specific for Pcdha1, Pcdha10, Pcdha12, Pcdhac1, Pcdhac2, and Purkinje cell protein 2 (Pcp2) (41). The sections were then photographed with a BIOREVO BZ-9000 microscope (Keyence).…”

Upstream Elements Involved in Vivo in Activation of the Brain-specific Rat Aldolase C Gene

“…Some genes are highly specific to certain neuron subpopulations (1,2), whereas others, especially isoenzymes of the general metabolism, are expressed in any types of neurons, although at different levels (3,4). The aldolase C gene, coding for the brain-specific form of the glycolytic enzyme aldolase, belongs to this last class and, in addition, is also transcribed in glial cells (5,6).…”

“…Like other genes broadly expressed in the brain, the aldolase C gene exhibits several features of housekeeping genes, namely the absence of canonical TATA and CAAT boxes, GC-rich sequence, and multiple transcription start sites (3,4,(7)(8)(9). We have previously reported that brain-specific expression of the CAT 1 reporter gene in transgenic mice could be directed by a short (Ͻ115 bp) promoter fragment of the aldolase C gene (10); however, this short fragment was not able to direct the high level transgene expression observed with the full-length 5.5-kb 5Ј-flanking region (11,12). To identify DNA regulatory elements required to ensure high level gene expression in the brain, we have now defined in vivo, in transgenic mice, activating sequences of the aldolase C gene and some of the cognate DNA-binding transcription factors.…”