Muc4/sialomucin complex,the intramembrane ErbB2 ligand,translocates ErbB2 to the apical surface in polarized epithelial cells

Muc4/Sialomucin complex (SMC) acts as an intramembrane ligand for the receptor tyrosine kinase ErbB2, inducing a limited phosphorylation of the receptor. Because Muc4/SMC is found at the apical surfaces of polarized epithelial cells and ErbB2 is often basolateral, the question arises as to whether these components become associated in polarized cells. To address this question, we examined the localization of these proteins in polarized human colon carcinoma CACO-2 cells. Dual color immunofluorescence analysis by confocal microscopy demonstrated the basolateral localization of the ErbB2 in these cells; it is primarily co-localized with E-cadherin at adherens junctions. Expression of apical Muc4/SMC in these cells by transient transfection results in the localization of the ErbB2 at the apical surface. Two-color confocal microscopy indicated that ErbB2 is colocalized with Muc4/SMC in the transfected cells but not in untransfected cells in the same culture. The change of localization of ErbB2 was confirmed by cell surface biotinylation of apical and basolateral proteins, followed by streptavidin precipitation and the subsequent detection of ErbB2 by immunoblotting. In contrast, Na+/K+-ATPase maintains its basolateral localization in Muc4/SMC-transfected cells, indicating that the translocation of ErbB2 is not the result of depolarization of the cells. A potential physiological role for the apical localization of ErbB2 is indicated by the fact that ErbB2 phosphorylated at tyrosine 1248 is found predominantly in Muc4/SMC-transfected cells, but not in untransfected cells, and is co-localized with the apical Muc4/SMC. The ability of Muc4/SMC to alter the localization of ErbB2, particularly a phosphorylated form of it, in epithelial cells, suggests that it has an important role in regulating ErbB2 signaling.     

Down syndrome and maternal age: the effect of erroneous assignment of parental origin.

It is tempting to assume that the maternal age effect in trisomy 21 is confined to cases arising from errors of maternal gametogenesis. However, it has been suggested that this hypothesis is incompatible with the results of studies, based on the subjective assessment of chromosome polymorphisms, of the parental origin of the additional chromosome. Contrary to the hypothesis, these studies appear to indicate that the ratio of maternal to paternal errors does not depend significantly on maternal age. I show here that the hypothesis need not be rejected if the proportion of published parental assignments that are incorrect is greater than or equal to 8%, a figure regarded as realistic by some experienced cytogeneticists.     

A study of genetic linkage heterogeneity in 35 adult-onset polycystic kidney disease families

A genetic heterogeneity analysis of 35 kindreds with adult-onset polycystic kidney disease (ADPKD) was carried out using the D16S85, D16S84, D16S125 and D16S94 loci that are closely linked to the PKD1 locus on chromosome 16. The results show that the likelihood of two ADPKD loci is 2,514.9 times greater than for a single locus (P < 0.0001). The maximum likelihood lod score is 27.38 under heterogeneity with PKD1 lying 4.9 cM proximal to D16S85 (in males). At least 3% of kindreds are unlinked to PKD1, since the 95% confidence limits of alpha, the proportion of families linked to PKD1, are 0.54–0.97. Only 2 out of 35 kindreds (5.7%) show statistically significant evidence of non-linkage to PKD1, with conditional probabilities of 0.987 and 0.993 that the disease locus is unlinked. This confirms the existence of a small subgroup of ADPKD kindreds that are unlinked to PKD1 and provides a firm basis for genetic counselling of this population on the basis of DNA probes.     

Linkage of a major breast cancer gene to chromosome 17q12-21: results from 15 Edinburgh families.

DNA from members of 15 pedigrees each containing between three and eight cases of breast cancer have been collected from southeastern Scotland. Polymorphic markers on chromosome 17q were screened to locate a putative breast cancer gene by using DNA from relevant individuals within these families. Pairwise LOD scores were calculated for markers D17S74, NM23, D17S588, and D17S579. The maximal summated LOD for the 15 families was 5.44 at theta = .034, when D17S588 (42D6) was used. In these breast cancer families, a subset which did not give evidence for linkage to this region could be identified.     

An efficient multiple-cell approach to automatic aneuploidy screening

We describe a statistical method of discriminating efficiently, on the basis of multiple-cell measurements without operator interaction, between chromosomally normal human cell lines and those either containing a single additional chromosome or missing one chromosome. We begin by defining hypothetical but realistic "confusion matrices," which give the probabilities of (1) assigning each chromosome to each of various possible groups and (2) rejecting it as unclassifiable. From these, false-positive and false-negative rates of 0.01 and 0.001, respectively, are found to be attainable by processing 16 to 32 cells if the average probability of misclassifying or rejecting individual chromosomes is 5% to 9% for "Denver" groups or 10% to 17% for homologous pairs. Since these values are probably within the reach of current technology, the method is a basis for a realistic, fully automatic screening system. We also show how the method can be extended to the detection of quite general types of chromosomally abnormal cell lines.     

Effect of trypsinization on thiamine transport in Escherichia coli Crookes

Trypsin treatment of intact $\text{Escherichia coli}$ Crookes cells is a probe for analyzing the functioning of protein sites in thiamine transport. These sites are cryptic since cells with different levels of thiamine transport activity respond differently to trypsin treatment. Mild trypsin treatment of cells with normal transport activity enhanced the velocity of uptake and decreased the capacity; more rigorous treatment reduced both parameters. Cells with low activity showed greatly increased rates of uptake and capacities under all but the mildest treatment conditions. These observations are consistent with a trypsin unmasking of thiamine transport sites in low activity cells and a destruction of the sites in higher activity cells.     

ULTRASTRUCTURAL STUDIES OF SPERMATOGENESIS IN THE ANTHOCEROTALES. II. THE BLEPHAROPLAST AND ANTERIOR MITOCHONDRION IN PHAEOCEROS LAEVIS: LATER DEVELOPMENT

Chromatin condensation begins as the multilayered structure (MLS) in Phaeoceros reaches its maximally structured differentiation. As nuclear elongation and chromatin condensation proceed, the S_2–4 strata disappear, and the nuclear beak extends between the S₁ and the nearly spherical anterior mitochondrion. In a mature sperm the mitochondrion is elongate and lies completely anterior to the blunt front end of the nucleus. The 12 S₁ tubules extend over the anterior mitochondrion and nucleus, but their number becomes reduced to five at the level where the nucleus' midportion is constricted. The anterior ends of the S₁ tubules lie embedded in a rather conical osmiophilic crest. Flagellar insertion is restricted to the extreme anterior tip of the S₁ tubules. The locomotory apparatus in Phaeoceros is compared with that of other bryophytes.     

OBSERVATIONS ON THE PROCAMBIUM AND PRIMARY PHLOEM OF PELARGONIUM DOMESTICUM

Carothers , Zane B. (U. Kentucky, Lexington.) Observation on the procambium and primary phloem of Pelargonium domesticum. Amer. Jour. Bot. 46(6): 397–404. Illus. 1959.—Stems of Pelargonium domesticum, a shrubby member of the Geraniaceae, were studied ontogenetically. It was found that a continuous cylinder of procambium is formed within the terminal millimeter of the young shoot. The pattern of trace procambium within this region is illustrated. Three traces are associated with each leaf; nodes are trilacunar. The prominent, sclerous “pericycle,” a diagnostic character of the family, actually consists of protophloem fibers. Commonly septate, these cells may contain as many as 4 nucleate protoplasts. The fibers average 1177 μ in length and 31 μ in diameter. Fiber walls average 5 μ in thickness, the inner pit apertures 6 μ in length. Metaphloem sieve tube elements averaged 203 μ in length.     

OBSERVATIONS ON THE PHI-THICKENINGS AND CASPARIAN STRIPS IN PELARGONIUM ROOTS

An ultrastructural examination of hypodermal cell walls failed to demonstrate the presence of casparian strips as has been reported in the literature. Rather, these cells have supportive phi-thickenings which differ significantly from casparian strips. Adjacent phi-thickenings are convex-convex and stratified in transection, with an uneven surface like the rest of the phi-cell's wall. They may be unbranched or branched and in the latter case, especially, they may appear on tangential as well as radial and transverse walls. They are lignified early in development and are associated with microtubules which lie parallel to the long axis of the thickening. Further, the plasmalemma exhibits no special adherence to the thickened wall region. In each of these and other characteristics phi-thickenings contrast sharply with casparian strips.