Gene ecology: a cis-acting gene-to-gene interaction due to the spatial arrangement of genes in chromosomes affects neighbouring transfected c-H-ras expression.

A cis-acting interference between gene activities, which occurs when two genes lie on the same DNA strand and have an intergenic distance less than a defined length, was previously deduced when chromosomal organizations of various higher eukaryote nuclear genes in clusters were compared. In order to investigate such an interference due to arrangement of genes along chromosomes, we have isolated a few cell lines which possessed (i) human mutated c-H-ras fused with the mouse mammary tumour virus long terminal repeat and (ii) the E. coli xanthine-guanine phosphoribosyltransferase (gpt) gene with the SV40 promoter, on the same or on different DNA strands, separated by a short intergenic distance or unlinked. Since the cancerous phenotype of a cell can be readily identified due to c-H-ras expression, we examined in these cell lines whether continuous c-H-ras expression, induced by dexamethasone, is disturbed through a cis-acting gene-to-gene interaction when the expression of the neighbouring gpt gene is enforced and as a result, the cancerous state of a cell is converted to the 'normal' state. The enforced expression of the neighbouring gpt gene was shown to alter c-H-ras expression, and thus reversible conversion of a cell between cancerous and normal states occurred only when the cell possessed an optimum number of the gene pair, in which both c-H-ras and the gpt gene were on the same DNA strand. This implies that the spatial arrangement of genes in chromosomes plays an important role in the regulation of gene expression in a cluster.

Morphology of the kidney of the platypus (Ornithorhynchus anatinus: Monotremata).

The platypus kidney shows morphological similarities to those of other mammals. Macroscopically, the cortex is easily distinguishable from the fairly wide medulla. Within the medulla, no clear border is observed between the inner and outer zones. Light and transmission electron microscopically, the glomeruli show quite similar architecture to those of other mammals; however, the glomerular lobulation is very clear. The glomerular tufts are rather simple, but capillary lumen varies widely in size, which is one of the unique features of the platypus kidney. The urinary tubule is generally similar to that of human and other mammals in shape and segmentation; however, the staining specificities of histochemical reactions and the shape of epithelial cells of the Henle's loop differ from those of other mammals. The most conspicuous features are: 1) although no protein casts are found in the tubular lumina, epithelial cells of the proximal convoluted tubule (PCT) have numerous electron-dense vesicles as in human nephrotic kidneys; and 2) the platypus Henle's loop consists of the thick epithelial cells similar to the mammalian type nephron of birds. As compared to those of other mammals such as humans and rats, our observations suggest that the platypus kidney is less developed, in terms of evolution.

An optimum dose of c-H-ras is a prerequisite for hormone-dependent conversion of a cell between cancerous and normal states in tissue culture.

We have established a few cell lines which can be reversibly converted from cancerous to normal and vice versa by the addition to, or removal from the culture medium of glucocorticoid hormone. These cell lines were derived from mouse NIH 3T3 cells and possessed the integrated gene pairs on chromosomes, which are composed of human mutated c-H-ras fused with mouse mammary tumour virus long terminal repeat and E. coli xanthine-guanine phosphoribosyltransferase gene with the SV40 promoter. We have characterised these cell lines in order to elucidate an essential requirement for the conversion of the state of a cell. It was found that the presence of at least two to three copies of the gene pair per diploid genome are essential. An approximate threshold level of c-H-ras 1.6 kb RNA required for reversible conversion was estimated.

Intestinal mucosa and intra-abdominal lymphoid tissues of the platypus, Ornithorhynchus anatinus.

The morphological features of the intestinal mucosa and intra-abdominal lymphoid tissues of the platypus were examined. The mucosal surface of the intestine was characterized by the formation of large folds instead of the finger-like villi found in placental mammals. The lamina propria of the mucosal fold was well developed and contained numerous lymphocytes, expressing the lymphoid nature which is characteristic of the lamina propria of mammalian intestines. Although numerous well-developed Peyer's patches were observed in the ileum, solitary lymphoid nodules could not be found anywhere in the small intestine. Other intra-abdominal lymphoid tissues, particularly mesenteric lymphoid nodules, were well developed. However, each nodule represented a single follicle in contrast to the mammalian mesenteric lymph node which is composed of numerous follicles fused together. On the basis of the above findings, the tissues in question are considered to be at an evolutionary level preceding that of placental mammals.