Differential expression of the human mucin genes MUC1 to MUC5 in relation to growth and differentiation of different mucus-secreting HT-29 cell subpopulations

1993 ◽  
Vol 106 (3) ◽  
pp. 771-783 ◽  
Author(s):  
T. Lesuffleur ◽  
N. Porchet ◽  
J.P. Aubert ◽  
D. Swallow ◽  
J.R. Gum ◽  
...  
Keyword(s):  
Steady State ◽  
Apical Surface ◽  
Parental Population ◽  
Mucin Expression ◽  
Mucin Genes ◽  
Muc Genes ◽  
Cell Subpopulations ◽  
Two Populations ◽  
Ht 29 ◽  
Muc1 Glycoprotein

Mucin expression was analysed, in relation to cell growth, in parental HT-29 cells and in two populations of mucus-secreting HT-29 cells selected by adaptation to methotrexate (HT29-MTX) or 5-fluorouracil (HT29-FU). These two populations express mature mucins that differ in their immunoreactivity to antibodies against gastric (HT29-MTX) or colonic mucins (HT29-FU). In the parental population, at late confluency, only very few cells produce mucins or the MUC1 glycoprotein, this being consistent with the low level of expression of the mRNAs corresponding to the MUC1 to MUC5C mucin genes. In the HT29-MTX and HT29-FU populations, the appearance of mucus droplets, as shown by histochemistry and immunofluorescence, starts a few days after confluency, progressively involving a greater proportion of cells and reaching a steady state at late confluency. The MUC1 glycoprotein appears earlier, already being detectable in preconfluent cells. Its distribution is restricted to the apical surface of the cells and is distinct from that of the mucus droplets. In both populations the growth-related levels of MUC1 mRNA are concordant with the apparent levels of expression of the MUC1 glycoprotein. The levels of MUC2, MUC3, MUC4 and MUC5C mRNAs differ from one population to another and, within each population, according to the stage of the culture. The highest levels of MUC2 and MUC4 mRNAs are found in the HT29-FU cells, whereas the highest levels of MUC3 and MUC5C are found in the HT29-MTX cells, suggesting that the differences observed in the mature mucins expressed by either population may be related to which MUC genes are expressed. In both populations significant or even high levels of MUC mRNAs are already present in early cultures, i.e. at a stage when the mature mucins are not yet detectable, suggesting that mucin maturation is a later event.

Expression of the human trefell factor 3 (TFF3) in relation to growth and differentiation of HT-29 cell subpopulations

2001 ◽  
Vol 120 (5) ◽  
pp. A508-A508
Author(s):  
S DURUAL ◽  
F MORO ◽  
V GOUYER ◽  
C BLANCHARD ◽  
C LABOISSE ◽  
...  
Keyword(s):  
Cell Subpopulations ◽  
Ht 29

scholarly journals Ingested Engineered Nanomaterials Affect the Expression of Mucin Genes—An In Vitro-In Vivo Comparison

Nanomaterials ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 2621
Author(s):  
Gerrit Bredeck ◽  
Angela A. M. Kämpfer ◽  
Adriana Sofranko ◽  
Tina Wahle ◽  
Veronika Büttner ◽  
...  
Keyword(s):  
Toxicity Testing ◽  
In Vitro Models ◽  
Engineered Nanomaterials ◽  
Intestinal Mucus ◽  
Specific Effects ◽  
Intestinal Infections ◽  
Mucin Expression ◽  
Mucin Genes

The increasing use of engineered nanomaterials (ENM) in food has fueled the development of intestinal in vitro models for toxicity testing. However, ENM effects on intestinal mucus have barely been addressed, although its crucial role for intestinal health is evident. We investigated the effects of ENM on mucin expression and aimed to evaluate the suitability of four in vitro models of increasing complexity compared to a mouse model exposed through feed pellets. We assessed the gene expression of the mucins MUC1, MUC2, MUC5AC, MUC13 and MUC20 and the chemokine interleukin-8 in pre-confluent and confluent HT29-MTX-E12 cells, in stable and inflamed triple cultures of Caco-2, HT29-MTX-E12 and THP-1 cells, and in the ileum of mice following exposure to TiO2, Ag, CeO2 or SiO2. All ENM had shared and specific effects. CeO2 downregulated MUC1 in confluent E12 cells and in mice. Ag induced downregulation of Muc2 in mice. Overall, the in vivo data were consistent with the findings in the stable triple cultures and the confluent HT29-MTX-E12 cells but not in pre-confluent cells, indicating the higher relevance of advanced models for hazard assessment. The effects on MUC1 and MUC2 suggest that specific ENM may lead to an elevated susceptibility towards intestinal infections and inflammations.

Studies of Muc-1 mucin expression and polarity in the mouse mammary gland demonstrate developmental regulation of Muc-1 glycosylation and establish the hormonal basis for mRNA expression

1992 ◽  
Vol 101 (1) ◽  
pp. 191-199 ◽  
Author(s):  
G. Parry ◽  
J. Li ◽  
J. Stubbs ◽  
M.J. Bissell ◽  
C. Schmidhauser ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cdna Probe ◽  
Mouse Mammary Gland ◽  
Mammary Development ◽  
Mammary Epithelial ◽  
Mrna Levels ◽  
Apical Surface ◽  
Ductal Cells ◽  
Carbohydrate Epitopes ◽  
Mucin Expression

Muc-1 is a major mucin glycoprotein expressed on the surface of mammary epithelial cells. It has attracted considerable attention as it is expressed in an aberrant form on many breast tumor cells. Here we describe studies using a recently obtained cDNA probe of Muc-1 expression during lactogenic development in the mouse. Northern blot analysis demonstrated that Muc-1 is expressed at all stages of lactogenic development but its levels are increased very significantly during mid-pregnancy and into lactation. The basis of this was examined using CID-9 mammary epithelial cell cultures. It was found that in the presence of insulin Muc-1 mRNA levels were increased by both hydrocortisone and prolactin, with the combination of the three hormones supporting maximum expression. Muc-1 mRNA levels were also modulated by culturing cells on a basement-membrane-like extracellular matrix that promoted mRNA levels 5- to 10-fold above levels in cells cultured on plastic tissue culture dishes. Immunocytochemical studies using monoclonal antibodies to carbohydrate epitopes on Muc-1 demonstrated that while Muc-1 was found at all developmental stages, it became increasingly sialylated during the course of pregnancy and into lactation. Additionally, we found that while Muc-1 is tightly polarized to the apical surface of the epithelium of lactating and pregnant mice it exhibited a less-polarized distribution on a small proportion of ductal cells in virgin mice. We conclude that the expression of Muc-1 is regulated at several different levels and by a number of different factors. We speculate that this may reflect different functional roles for Muc-1 at different stages of mammary development.

scholarly journals Interactions between actin filaments and between actin filaments and membranes in quick-frozen and deeply etched hair cells of the chick ear.

1982 ◽  
Vol 95 (1) ◽  
pp. 249-261 ◽  
Author(s):  
N Hirokawa ◽  
L G Tilney
Keyword(s):  
Hair Cells ◽  
Actin Filaments ◽  
Apical Cell ◽  
Apical Surface ◽  
Contractile Ring ◽  
Vestibular Organ ◽  
Apical Cell Membrane ◽  
Cuticular Plate ◽  
Quick Freezing ◽  
Two Populations

Replicas of the apical surface of hair cells of the inner ear (vestibular organ) were examined after quick freezing and rotary shadowing. With this technique we illustrate two previously undescribed ways in which the actin filaments in the stereocilia and in the cuticular plate are attached to the plasma membrane. First, in each stereocilium there are threadlike connectors running from the actin filament bundle to the limiting membrane. Second, many of the actin filaments in the cuticular plate are connected to the apical cell membrane by tiny branched connecting units like a "crow's foot." Where these "feet" contact the membrane there is a small swelling. These branched "feet" extend mainly from the ends of the actin filaments but some connect the lateral surfaces of the actin filaments as well. Actin filaments in the cuticular plate are also connected to each other by finer filaments, 3 nm in thickness and 74 +/- 14 nm in length. Interestingly, these 3-nm filaments (which measure 4 nm in replicas) connect actin filaments not only of the same polarity but of opposite polarities as documented by examining replicas of the cuticular plate which had been decorated with subfragment 1 (S1) of myosin. At the apicolateral margins of the cell we find two populations of actin filaments, one just beneath the tight junction as a network, the other at the level of the zonula adherens as a ring. The latter which is quite substantial is composed of actin filaments that run parallel to each other; adjacent filaments often show opposite polarities, as evidenced by S1 decoration. The filaments making up this ring are connected together by the 3-nm connectors. Because of the polarity of the filaments this ring may be a "contractile" ring; the implications of this is discussed.

scholarly journals Differential mucin expression in colon carcinoma HT-29 clones with variable resistance to 5-fluorouracil and methotrexate

2004 ◽  
Vol 96 (2) ◽  
pp. 145-151 ◽  
Author(s):  
Emmanuelle Leteurtre ◽  
Valérie Gouyer ◽  
Karine Rousseau ◽  
Odile Moreau ◽  
Alain Barbat ◽  
...  
Keyword(s):  
Colon Carcinoma ◽  
Variable Resistance ◽  
Mucin Expression ◽  
Ht 29

scholarly journals Steady-state distribution and biogenesis of endogenous Madin-Darby canine kidney glycoproteins: evidence for intracellular sorting and polarized cell surface delivery.

1989 ◽  
Vol 109 (5) ◽  
pp. 2117-2127 ◽  
Author(s):  
M P Lisanti ◽  
A Le Bivic ◽  
M Sargiacomo ◽  
E Rodriguez-Boulan
Keyword(s):  
Steady State ◽  
Epithelial Cell ◽  
Cell Surface ◽  
Mdck Cells ◽  
Apical Surface ◽  
Steady State Distribution ◽  
State Distribution ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

We used domain-selective biotinylation/125I-streptavidin blotting (Sargiacomo, M., M. P. Lisanti, L. Graeve, A. Le Bivic, and E. Rodriguez-Boulan. 1989 J. Membr. Biol. 107:277-286), in combination with lectin precipitation, to analyze the apical and basolateral glycoprotein composition of Madin-Darby canine kidney (MDCK) cells and to explore the role of glycosylation in the targeting of membrane glycoproteins. All six lectins used recognized both apical and basolateral glycoproteins, indicating that none of the sugar moieties detected were characteristic of the particular epithelial cell surface. Pulse-chase experiments coupled with domain-selective glycoprotein recovery were designed to detect the initial appearance of newly synthesized glycoproteins at the apical or basolateral cell surface. After a short pulse with a radioactive precursor, glycoproteins reaching each surface were biotinylated, extracted, and recovered via precipitation with immobilized streptavidin. Several basolateral glycoproteins (including two sulfated proteins) and at least two apical glycoproteins (one of them the major sulfated protein of MDCK cells) appeared at the corresponding surface after 20-40 min of chase, but were not detected in the opposite surface, suggesting that they were sorted intracellularly and vectorially delivered to their target membrane. Several "peripheral" apical proteins were detected at maximal levels on the apical surface immediately after the 15-min pulse, suggesting a very fast intracellular transit. Finally, domain-selective labeling of surface carbohydrates with biotin hydrazide (after periodate oxidation) revealed strikingly different integral and peripheral glycoprotein patterns, resembling the Con A pattern, after labeling with sulfo-N-hydroxy-succinimido-biotin. The approaches described here should be useful in characterizing the steady-state distribution and biogenesis of endogenous cell surface components in a variety of epithelial cell lines.

Expression of the human trefell factor 3 (TFF3) in relation to growth and differentiation of HT-29 cell subpopulations

2001 ◽  
Vol 120 (5) ◽  
pp. A508
Author(s):  
Stephane Durual ◽  
Frederic Moro ◽  
Valerie Gouyer ◽  
Carine Blanchard ◽  
Christian L. Laboisse ◽  
...  
Keyword(s):  
Cell Subpopulations ◽  
Ht 29

Acyl-CoA: cholesterol acyltransferase in HT 29 cell subpopulations. Defect of activity in the undifferentiated cells

1991 ◽  
Vol 1085 (2) ◽  
pp. 265-271 ◽  
Author(s):  
Viviane Viallard ◽  
Christiane Lacombe ◽  
Christine Nebot ◽  
Isabelle Castan ◽  
Véronique Trocheris
Keyword(s):  
Cholesterol Acyltransferase ◽  
Undifferentiated Cells ◽  
Cell Subpopulations ◽  
Ht 29

scholarly journals Constitutive endocytosis and recycling of NKCC2 in rat thick ascending limbs

2010 ◽  
Vol 299 (5) ◽  
pp. F1193-F1202 ◽  
Author(s):  
Gustavo R. Ares ◽  
Pablo A. Ortiz
Keyword(s):  
Plasma Membrane ◽  
Steady State ◽  
Acute Treatment ◽  
Apical Membrane ◽  
Surface Expression ◽  
Membrane Cholesterol ◽  
Thick Ascending Limb ◽  
Apical Surface ◽  
Total Pool ◽  
Stimulation Of

The Na-K-2Cl cotransporter (NKCC2) mediates NaCl absorption by the thick ascending limb of Henle's loop (THAL). Exocytosis and endocytosis regulates surface expression of most transporters. However, little is known about the mechanism of NKCC2 trafficking in the absence of stimulating hormones and whether this mechanism contributes to regulation of steady-state surface expression of apical NKCC2 in the THAL. We tested whether NKCC2 undergoes constitutive endocytosis that regulates steady-state surface NKCC2 and NaCl reabsorption in THALs. We measured steady-state surface NKCC2 levels and the rate of NKCC2 endocytosis by surface biotinylation and Western blot and confocal microscopy of isolated perfused rat THALs. We observed constitutive NKCC2 endocytosis over 30 min that averaged 21.5 ± 2.7% of the surface pool. We then tested whether methyl-β-cyclodextrin (MβCD), a compound that inhibits endocytosis by chelating membrane cholesterol, blocked NKCC2 endocytic retrieval. We found that 30-min treatment with MβCD (5 mM) blocked NKCC2 endocytosis by 81% ( P < 0.01). Blockade of endocytosis by MβCD induced accumulation of NKCC2 at the apical membrane as demonstrated by a 60 ± 16% ( P < 0.05) increase in steady-state surface expression and enhanced apical surface NKCC2 immunostaining in isolated, perfused THALs. Acute treatment with MβCD did not change the total pool of NKCC2. MβCD did not affect NKCC2 trafficking when it was complexed with cholesterol before treatment. Inhibition endocytosis with MβCD enhanced NKCC2-dependent NaCl entry by 57 ± 16% ( P < 0.05). Finally, we observed that a fraction of retrieved NKCC2 recycles back to the plasma membrane (36 ± 7%) over 30 min. We concluded that constitutive NKCC2 trafficking maintains steady-state surface NKCC2 and regulates NaCl reabsorption in THALs. These are the first data showing an increase in apical membrane NKCC2 in THALs by altering the rates of constitutive NKCC2 trafficking, rather than by stimulation of hormone-dependent signaling.

scholarly journals Inferring inter- and intrachromosomal Dobzhansky-Muller incompatibilities from imbalanced segregation of recombinant haplotypes in hybrid populations

2021 ◽  
Author(s):  
Juan Li ◽  
Molly Schumer ◽  
Claudia Bank
Keyword(s):  
Genetic Mapping ◽  
Low Power ◽  
Reproductive Isolation ◽  
Statistical Tests ◽  
Parental Population ◽  
Swordtail Fish ◽  
Haplotype Frequencies ◽  
New Statistics ◽  
Negative Epistasis ◽  
Two Populations

Dobzhansky-Muller incompatibilities (DMIs) are a major component of reproductive isolation between species. DMIs imply negative epistasis, exposed when two diverged populations hybridize. Mapping the locations of DMIs has largely relied on classical genetic mapping, but these approaches are stymied by low power and the challenge of identifying DMI loci on the same chromosome, because strong initial linkage of parental haplotypes weakens statistical tests. Here, we propose new statistics to infer negative epistasis from haplotype frequencies in hybrid populations. When two divergent populations hybridize, the variance of two-locus heterozygosity decreases faster with time at DMI loci than at random pairs of loci. If two populations hybridize at near-even admixture proportions, the deviation of the observed variance from its expectation is negative, which enables us to detect signals of intermediate to strong negative epistasis both within and between chromosomes. When the initial proportion of the two parental populations is uneven, only strong DMIs can be detected with our method, unless migration reintroduces haplotypes from the minor parental population. We use the two new statistics to infer candidate DMIs from three hybrid populations of swordtail fish. We identify numerous new DMI candidates some of which are inferred to interact with several loci within and between chromosomes. Moreover, we discuss our results in the context of an expected enrichment in intrachromosomal over interchromosomal DMIs.

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