Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells.

In situ hybridization of human chromosome 18 and X-specific alphoid DNA-probes was performed in combination with three dimensional (3D) and two dimensional (2D) image analysis to study the interphase distribution of the centric heterochromatin (18c and Xc) of these chromosomes in cultured human cells. 3D analyses of 18c targets using confocal laser scanning microscopy indicated a nonrandom disposition in 73 amniotic fluid cell nuclei. The shape of these nuclei resembled rather flat cylinders or ellipsoids and targets were preferentially arranged in a domain around the nuclear center, but close to or associated with the nuclear envelope. Within this domain, however, positionings of the two targets occurred independently from each other, i.e., the two targets were observed with similar frequencies at the same (upper or lower) side of the nuclear envelope as those on opposite sides. This result strongly argues against any permanent homologous association of 18c. A 2D analytical approach was used for the rapid evaluation of 18c positions in over 4000 interphase nuclei from normal male and female individuals, as well as individuals with trisomy 18 and Bloom's syndrome. In addition to epithelially derived amniotic fluid cells, investigated cell types included in vitro cultivated fibroblastoid cells established from fetal lung tissue and skin-derived fibroblasts. In agreement with the above 3D observations 18c targets were found significantly closer (P less than 0.01) to the center of the 2D nuclear image (CNI) and to each other in all these cultures compared to a random distribution derived from corresponding ellipsoid or cylinder model nuclei. For comparison, a chromosome X-specific alphoid DNA probe was used to investigate the 2D distribution of chromosome X centric heterochromatin in the same cell types. Two dimensional Xc-Xc and Xc-CNI distances fit a random distribution in diploid normal and Bloom's syndrome nuclei, as well as in nuclei with trisomy X. The different distributions of 18c and Xc targets were confirmed by the simultaneous staining of these targets in different colors within individual nuclei using a double in situ hybridization approach.

Chromosomal organization and localization of the human urokinase inhibitor gene: perfect structural conservation with ovalbumin.

Plasminogen activator inhibitor 2 (PAI-2) plays an essential role in the regulation of localized extracellular proteolysis by its inactivation of urokinase. Using probes derived from a cDNA we isolated from lipopolysaccharide (LPS)-stimulated human peripheral blood monocytes, we have mapped, isolated, and determined the molecular organization of the gene for PAI-2 (PLANH2). In situ hybridization of the cDNA to normal metaphase chromosomes has confirmed our prior assignment of the gene for PAI-2 to chromosome 18 and further localized it to the long arm at 18q21.2-18q22. We have isolated nine independent genomic clones, two of which were found to contain the entire PAI-2 transcriptional unit of approximately 16.4 kilobase pairs (kbp). Analysis of the gene organization by restriction enzyme mapping, Southern blotting, and DNA sequencing revealed that the cDNA sequence is divided among eight exons interrupted by seven introns, the junctions of which all conform to the "GT-AG" consensus rule. In common with the arrangement found throughout, the serpin superfamily, of which PAI-2 is a member, the first intron is located just 5' to the initiator methionine residue, and the 3' untranslated region (UTR) is not interrupted by a splice junction. Determination of the transcription initiation site by primer extension analysis of monocytic mRNA indicated that our PAI-2 cDNA was, at most, only three nucleotides short of full length, yielding a primary PAI-2 transcript with a 66-bp first exon. A promoter "TATAAAbox" is located 30 bp upstream of the "cap" site.(ABSTRACT TRUNCATED AT 250 WORDS)

Prenatal diagnosis of a case of 46,XY,18p-/46,XY,18p+ mosaicism.

A case of mosaicism involving structural abnormality of chromosome 18 found in cultured amniotic fluid is reported.