Mapping of the SWAP70 gene to mouse chromosome 7 and human chromosome 11p15.

The protein SWAP-70 was isolated as part of a DNA recombination complex in B lymphocytes, where it is predominantly expressed. In resting B cells, SWAP-70 is found in the cytoplasm; upon B-cell activation, it is transported both into the nucleus and to the cell membrane, where it is associated with the B-cell receptor complex and may play a role in signal transduction. In the nucleus, its involvement in heavy-chain class switch recombination has been suggested. In this report, using restriction fragment length polymorphism, simple sequence length polymorphism, and fluorescence in situ hybridization, we map the chromosomal localization of the mouse and the human genes to syntenic regions of mouse mid Chromosome (Chr) 7 and human Chr 11p15.

The secretory phospholipase A2 gene is a candidate for the Mom1 locus, a major modifier of ApcMin-induced intestinal neoplasia.

Mutations in the APC gene are responsible for various familial and sporadic colorectal cancers. Min mice carry a dominant mutation in the homolog of the Apc gene and develop multiple adenomas throughout their small and large intestine. Quantitative trait loci studies have identified a locus, Mom1, which maps to the distal region of chromosome 4, that dramatically modifies Min-induced tumor number. We report here the identification of a candidate gene for Mom1. The gene for secretory type II phospholipase A2 (Pla2s) maps to the same region that contains Mom1 and displays 100% concordance between allele type and tumor susceptibility. Expression and sequence analysis revealed that Mom1 susceptible strains are most likely null for Pla2s activity. Our results indicate that Pla2s acts as a novel gene that modifies polyp number by altering the cellular microenvironment within the intestinal crypt.

A high-resolution linkage map of the lethal spotting locus: a mouse model for Hirschsprung disease.

Mice homozygous for the lethal spotting (ls) mutation exhibit aganglionic megacolon and a white spotted coat owing to a lack of neural crest-derived enteric ganglia and melanocytes. The ls mutation disrupts the migration, differentiation, or survival of these neural crest lineages during mammalian development. A human congenital disorder, Hirschsprung disease (HSCR), is also characterized by aganglionic megacolon of the distal bowel and can be accompanied by hypopigmentation of the skin. HSCR has been attributed to multiple loci acting independently or in combination. The ls mouse serves as one animal model for HSCR, and the ls gene may represent one of the loci responsible for some cases of HSCR in humans. This study uses 753 N2 progeny from a combination of three intersubspecific backcrosses to define the molecular genetic linkage map of the ls region and to provide resources necessary for positional cloning. Similar to some cases of HSCR, the ls mutation acts semidominantly, its phenotypic effects dependent upon the presence of modifier genes segregating in the crosses. We have now localized the ls mutation to a 0.8-cM region between the D2Mit113 and D2Mit73/D2Mit174 loci. Three genes, endothelin-3 (Edn3), guanine nucleotide-binding protein alpha-stimulating polypeptide 1 (Gnas), and phosphoenolpyruvate carboxykinase (Pck1) were assessed as candidates for the ls mutation. Only Edn3 and Gnas did not recombine with the ls mutation. Mutational analysis of the Edn3 and Gnas genes will determine whether either gene is responsible for the neural crest deficiencies observed in ls/ls mice.

Differential isoactin gene expression in the sphincteric and nonsphincteric gastrointestinal smooth muscles of the opossum.

The sphincteric smooth muscle tissues of the gastrointestinal tract have been shown to possess mechanical properties distinct from the surrounding nonsphincteric smooth muscle tissues. Little is currently known regarding the molecular basis of this differential smooth muscle development and function. Actin is an important contractile protein whose expression has been linked to the normal development and function of smooth muscle tissues. The purpose of this study was to characterize isoactin gene expression in the sphincteric versus nonsphincteric smooth muscle tissues of the gastrointestinal tract. Northern blot analysis was performed on manometrically identified sphincteric and the flanking nonsphincteric smooth muscle tissues of the adult opossum. Quantitative analysis revealed a distinct pattern of isoactin gene expression in the sphincteric versus nonsphincteric smooth muscle tissues of the gut. The sphincteric smooth muscle tissues expressed significantly lower total quantities of isoactin mRNA than their surrounding nonsphincteric smooth muscle tissues. It is hypothesized that these differential patterns of isoactin gene expression may play a significant role in establishing the myogenic potential of the functionally distinct sphincteric and nonsphincteric smooth muscle tissues of the gut.

Heterogeneous isoactin gene expression in the adult rat gastrointestinal tract.

BACKGROUND: Normal gastrointestinal development is a complex process involving the precise integration of multiple cell types. To gain a better understanding of these processes, the present study examined isoactin gene expression in the adult rat gastrointestinal tract. METHODS: Northern blot analysis was performed on specified segments of the adult rat esophagus, stomach, small intestine, cecum, colon, rectum, and anus using actin isoform-specific complementary DNAs for all six vertebrate isoactins. RESULTS: Smooth muscle and cytoplasmic isoactins were heterogeneously coexpressed in a segment-specific manner throughout the gastrointestinal tract. In addition, striated muscle isoactin expression was also detected in segments of the adult rat esophagus, stomach, colon, cecum, rectum, and anus. Histological analysis indicated that the adult rat esophagus, stomach, and anus contained significant quantities of skeletal muscle, providing a source for the striated muscle isoactins detected in these gut segments. A similar source of striated muscle isoactin expression in the cecum, colon, and rectum was not identified. Both coordinate and independent regulation of isoactin gene expression was observed in the gastrointestinal tract, although distinct patterns of autoregulation were absent. CONCLUSIONS: This study represents the first complete analysis of isoactin gene expression in the adult rat gastrointestinal tract and provides the basis for future studies designed to investigate the factors responsible for these processes.

Altered isoactin gene expression in the affected bowel segments of the lethal spotted mouse.

BACKGROUND: Actin is a key contractile protein associated with the normal differentiation and function of gastrointestinal smooth muscle cells. Distinct changes in gastrointestinal smooth muscle cell morphology and function have been reported for the aganglionic rectum and megacolon of the adult lethal spotted mouse. This study examines what effect these changes in smooth muscle cell morphology and function have on the expression of the actin multigene family in both the aganglionic rectum and megacolon of the lethal spotted mouse. METHODS: Expression of the smooth muscle and cytoplasmic isoactins was examined by Northern blot analysis of the aganglionic rectum and megacolon of the homozygotic lethal spotted mouse and the equivalent bowel segments of control animals. RESULTS: The megacolon of the lethal spotted mouse showed a significant increase in gamma-smooth muscle isoactin expression. The aganglionic rectum of the lethal spotted mouse displayed a complex pattern of altered isoactin gene expression that included changes in both gamma-smooth muscle and beta-cytoplasmic isoactin expression. Strain-specific differences in the quantitative levels of isoactin gene expression were observed for the various bowel segments examined in this study. CONCLUSIONS: These results show that the changes in smooth muscle cell morphology and function observed in the lethal spotted mutant mouse are accompanied by significant alterations in isoactin gene expression.