scholarly journals Knockdown of p180 Eliminates the Terminal Differentiation of a Secretory Cell Line

2009 ◽  
Vol 20 (2) ◽  
pp. 732-744 ◽  
Author(s):  
Payam Benyamini ◽  
Paul Webster ◽  
David I. Meyer
Keyword(s):  
Cell Line ◽  
Mammalian Cells ◽  
Secretory Cell ◽  
Terminal Differentiation ◽  
Secretory Cells ◽  
Rnai Technology ◽  
Apo E ◽  
Secretory Phenotype ◽  
Rough Er ◽  
Necessary And Sufficient

We have previously reported that the expression in yeast of an integral membrane protein (p180) of the endoplasmic reticulum (ER), isolated for its ability to mediate ribosome binding, is capable of inducing new membrane biogenesis and an increase in secretory capacity. To demonstrate that p180 is necessary and sufficient for terminal differentiation and acquisition of a secretory phenotype in mammalian cells, we studied the differentiation of a secretory cell line where p180 levels had been significantly reduced using RNAi technology and by transiently expressing p180 in nonsecretory cells. A human monocytic (THP-1) cell line, that can acquire macrophage-like properties, failed to proliferate rough ER when p180 levels were lowered. The Golgi compartment and the secretion of apolipoprotein E (Apo E) were dramatically affected in cells expressing reduced p180 levels. On the other hand, expression of p180 in a human embryonic kidney nonsecretory cell line (HEK293) showed a significant increase in proliferation of rough ER membranes and Golgi complexes. The results obtained from knockdown and overexpression experiments demonstrate that p180 is both necessary and sufficient to induce a secretory phenotype in mammalian cells. These findings support a central role for p180 in the terminal differentiation of secretory cells and tissues.

scholarly journals Molecular analysis of ethyl methanesulfonate-induced reversion of a chromosomally integrated mutant shuttle vector gene in mammalian cells.

1988 ◽  
Vol 8 (10) ◽  
pp. 4185-4189 ◽  
Author(s):  
J A Greenspan ◽  
F M Xu ◽  
R L Davidson
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Dna Sequences ◽  
Mammalian Cells ◽  
Molecular Mechanisms ◽  
Shuttle Vector ◽  
Ethyl Methanesulfonate ◽  
Wild Type ◽  
Single Base ◽  
Type Sequence

The molecular mechanisms of ethyl methanesulfonate-induced reversion in mammalian cells were studied by using as a target a gpt gene that was integrated chromosomally as part of a shuttle vector. Murine cells containing mutant gpt genes with single base changes were mutagenized with ethyl methanesulfonate, and revertant colonies were isolated. Ethyl methanesulfonate failed to increase the frequency of revertants for cell lines with mutant gpt genes carrying GC----AT transitions or AT----TA transversions, whereas it increased the frequency 50-fold to greater than 800-fold for cell lines with mutant gpt genes carrying AT----GC transitions and for one cell line with a GC----CG transversion. The gpt genes of 15 independent revertants derived from the ethyl methanesulfonate-revertible cell lines were recovered and sequenced. All revertants derived from cell lines with AT----GC transitions had mutated back to the wild-type gpt sequence via GC----AT transitions at their original sites of mutation. Five of six revertants derived from the cell line carrying a gpt gene with a GC----CG transversion had mutated via GC----AT transition at the site of the original mutation or at the adjacent base in the same triplet; these changes generated non-wild-type DNA sequences that code for non-wild-type amino acids that are apparently compatible with xanthine-guanine phosphoribosyltransferase activity. The sixth revertant had mutated via CG----GC transversion back to the wild-type sequence. The results of this study define certain amino acid substitutions in the xanthine-guanine phosphoribosyltransferase polypeptide that are compatible with enzyme activity. These results also establish mutagen-induced reversion analysis as a sensitive and specific assay for mutagenesis in mammalian cells.

Structure and expression of ferritin genes in a human promyelocytic cell line that differentiates in vitro

1986 ◽  
Vol 6 (2) ◽  
pp. 566-573
Author(s):  
C C Chou ◽  
R A Gatti ◽  
M L Fuller ◽  
P Concannon ◽  
A Wong ◽  
...  
Keyword(s):  
Cell Line ◽  
Terminal Differentiation ◽  
Amino Acid Sequences ◽  
Multigene Families ◽  
Cdna Clones ◽  
Macrophage Differentiation ◽  
Time Points ◽  
Human Ferritin ◽  
Complex Manner

HL-60 is a human promyelocytic cell line with the capability of differentiating in vitro to give neutrophils, macrophages, or eosinophils. We screened libraries of HL-60 cDNA clones representing different time points during these differentiation processes to isolate clones corresponding to mRNAs whose expression is regulated during terminal differentiation. Upon sequencing this group of regulated clones, one clone encoding the heavy subunit and two clones encoding the light subunit of human ferritin were identified by reference to published amino acid sequences. Southern blot analyses showed that these clones are encoded by distinct multigene families. These clones identify two mRNAs whose ratios vary in a complex manner during both neutrophil and macrophage differentiation.

Dexamethasone regulation of terminal differentiation in 3T3-F442A preadipocyte cell line

1988 ◽  
Vol 1 (4) ◽  
pp. 285-293 ◽  
Author(s):  
Na�ma Mousta�d ◽  
Bernard Hainque ◽  
Annie Quignard-Boulang�
Keyword(s):  
Cell Line ◽  
Terminal Differentiation

scholarly journals Establishment and characterization of a human plasma cell myeloma culture having a rearranged cellular myc proto-oncogene

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1542-1549 ◽  
Author(s):  
AF Gazdar ◽  
HK Oie ◽  
IR Kirsch ◽  
GF Hollis
Keyword(s):  
Cell Line ◽  
Human Plasma ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Cultured Cells ◽  
Human Cell Line ◽  
Defined Medium ◽  
Chromosome 8 ◽  
Nuclear Antigen ◽  
Secretory Cells

Using a serum-free defined medium, we have established a human cell line, NCI-H929, from a malignant effusion occurring in a patient with IgAk myeloma. The cultured cells have the morphologic, ultrastructural, biochemical, immunologic, and cytochemical features of plasma cells. The cells have rearranged alpha and kappa genes and synthesize and secrete high amounts of IgAk (greater than 80 micrograms/10(6) cells per 24 hours). The cells express surface immunoglobulin (alpha and kappa), the plasma cell antigen PCA-1, the transferrin receptor (T9) and T10 but lack antigens associated with earlier stages of B cell development (HLA-DR, B1, B2, B4, CALLA), as well as other leukocyte- macrophage antigens and Epstein-Barr virus (EBV) nuclear antigen. Although molecular studies confirm that both the tumor and cultured cells are derived from the same clone of malignant B cells, the tumor cells were predominantly near-diploid, whereas the cultured cells are predominantly near-tetraploid with six copies of chromosome 8, four to six of which have an 8q + abnormality. However, both the tumor and the cultured cells have a rearrangement of the cellular c-myc proto- oncogene (located at 8q24) and express c-myc RNA. Although a modest number of human “plasmacytoid” cell lines have been established, most are lymphoblastoid lines lacking plasma cell features, while others appear to be early secretory cells. In contrast, NCI-H929 is a differentiated, highly secretory human plasma cell line.

scholarly journals Possible Arbovirus Found in Virome of Melophagus ovinus

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2375
Author(s):  
Alexander G. Litov ◽  
Oxana A. Belova ◽  
Ivan S. Kholodilov ◽  
Magomed N. Gadzhikurbanov ◽  
Larissa V. Gmyl ◽  
...  
Keyword(s):  
Weight Loss ◽  
Cell Line ◽  
Mammalian Cells ◽  
Skin Irritation ◽  
The Novel ◽  
Skin Injuries ◽  
Blood Sucking ◽  
Melophagus Ovinus ◽  
Porcine Embryo ◽  
The Republic

Members of the Lipopteninae subfamily are blood-sucking ectoparasites of mammals. The sheep ked (Melophagus ovinus) is a widely distributed ectoparasite of sheep. It can be found in most sheep-rearing areas and can cause skin irritation, restlessness, anemia, weight loss and skin injuries. Various bacteria and some viruses have been detected in M. ovinus; however, the virome of this ked has never been studied using modern approaches. Here, we study the virome of M. ovinus collected in the Republic of Tuva, Russia. In our research, we were able to assemble full genomes for five novel viruses, related to the Rhabdoviridae (Sigmavirus), Iflaviridae, Reoviridae and Solemoviridae families. Four viruses were found in all five of the studied pools, while one virus was found in two pools. Phylogenetically, all of the novel viruses clustered together with various recently described arthropod viruses. All the discovered viruses were tested on their ability to replicate in the mammalian porcine embryo kidney (PEK) cell line. Aksy-Durug Melophagus sigmavirus RNA was detected in the PEK cell line cultural supernate after the first, second and third passages. Such data imply that this virus might be able to replicate in mammalian cells, and thus, can be considered as a possible arbovirus.

scholarly journals Isolation of Human Transcripts Expressed in Hamster Cells from YACs by cDNA Representational Difference Analysis

1999 ◽  
Vol 9 (2) ◽  
pp. 182-188
Author(s):  
Jessie Gu ◽  
Xin-Yuan Guan ◽  
Melissa A. Ashlock
Keyword(s):  
Cell Line ◽  
Mammalian Cells ◽  
Positional Cloning ◽  
Yeast Artificial Chromosome ◽  
Expressed Sequence Tag ◽  
Gene Isolation ◽  
Representational Difference Analysis ◽  
Difference Analysis ◽  
Artificial Chromosomes ◽  
Link Type

Gene isolation methods used during positional cloning rely on physical contigs consisting of bacterial artificial chromosomes, P1, or cosmid clones. However, in most instances, the initial framework for physical mapping consists of contigs of yeast artificial chromosome (YACs), large vectors that are suboptimal substrates for gene isolation. Here we report a strategy to identify gene sequences contained within a YAC by using cDNA representational difference analysis (RDA) to directly isolate transcripts expressed from the YAC in mammalian cells. The RDA tester cDNAs were generated from a previously reported hamster cell line derived by stable transfer of a 590-kb YAC (911D5) that expressed NPC1, the human gene responsible for Niemann-Pick type C (NP-C). The driver cDNAs were generated from a control hamster cell line that did not contain the YAC that expressed NPC1. Among the gene fragments obtained by RDA,NPC1 was the most abundant product. In addition, two non-NPC1 fragments were isolated that were mapped to and expressed from 911D5. One of these RDA gene fragments (7-R) spans more than one exon and has 98% sequence identity with a human cDNA clone reported previously as an expressed sequence tag (EST), but not mapped to a chromosomal region. The other fragment (2-R) that had no significant sequence similarities with known mammalian genes or ESTs, was further localized to the region of overlap between YACs911D5 and 844E3. The latter YAC is part of a contig across the NP-C candidate region, but does not contain NPC1. This two-part approach in which stable YAC transfer is followed by cDNA RDA should be a useful adjunct strategy to expedite the cloning of human genes when a YAC contig is available across a candidate interval.[The sequence data described in this paper have been submitted to GenBank under accession nos. AF117641 and AF117642.]

Anatomy, ultrastructure, and functional morphology of the metathoracic tracheal defensive glands of the grasshopper Romalea guttata

1991 ◽  
Vol 69 (8) ◽  
pp. 2100-2108 ◽  
Author(s):  
Douglas W. Whitman ◽  
Johan P. J. Billen ◽  
David Alsop ◽  
Murray S. Blum
Keyword(s):  
Secretory Cell ◽  
Tactile Stimulation ◽  
Defensive Secretion ◽  
Smooth Endoplasmic Reticulum ◽  
Duct Cell ◽  
Secretory Cells ◽  
Glandular Tissue ◽  
Golgi Bodies ◽  
Romalea Guttata ◽  
Defensive Glands

In the lubber grasshopper Romalea guttata, the respiratory system produces, stores, and delivers a phenolic defensive secretion. The exudate is secreted by a glandular epithelium surrounding the metathoracic spiracular tracheal trunks. Embedded in the glandular tissue are multiple secretory units, each comprised of a basal secretory cell and an apical duct cell. Secretory cells have numerous mitochondria, a tubular, smooth endoplasmic reticulum, well-developed Golgi bodies, and a microvillilined vesicle thought to transfer secretion to the intracellular cuticular duct of a duct cell. Ducts empty into the metathoracic tracheal lumina where the exudate is stored behind the closed metathoracic spiracle. Tactile stimulation elicits secretion discharge, which begins when all spiracles except the metathoracic pair are closed and the abdomen is compressed. Increased hemostatic and pneumatic pressures drive air and secretion out of the spiracle with an audible hiss. Both metathoracic spiracles discharge simultaneously. The secretion erupts first as a dispersant spray, then as an adherent froth, and finally assumes the form of a slowly evaporating repellent droplet. Discharge force and number vary with eliciting stimuli, volume of stored secretion, and age, disturbance state, and temperature of the insect. Molting grasshoppers are unable to discharge because the stored exudate is lost with the shed cuticle. The advantages and limitations of a tracheal defensive system are discussed.

Sensitivity of mammary secretory cell turnover to protein restriction and re-alimentation during early lactation

1997 ◽  
Vol 1997 ◽  
pp. 130-130
Author(s):  
M.G. Goodwill ◽  
N.S. Jessop ◽  
J.D. Oldham
Keyword(s):  
Cell Proliferation ◽  
Mammary Gland ◽  
Milk Production ◽  
Secretory Cell ◽  
Protein Restriction ◽  
Secretory Cells ◽  
Early Lactation ◽  
Cell Turnover ◽  
Lactational Performance

Milk production depends on both the number and activity of secretory cells within the mammary gland. Our earlier work showed the sensitivity of lactational performance to changes in diet during lactation (Goodwill et al, 1996). This study investigated the influence of protein undernutrition and re-alimentation on secretory cell proliferation and death in the mammary gland of rats during early lactation.

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