lag-2 may encode a signaling ligand for the GLP-1 and LIN-12 receptors of C. elegans

Development ◽  
1994 ◽  
Vol 120 (10) ◽  
pp. 2913-2924 ◽  
Author(s):  
S.T. Henderson ◽  
D. Gao ◽  
E.J. Lambie ◽  
J. Kimble
Keyword(s):  
In Situ Hybridization ◽  
Membrane Protein ◽  
Fusion Protein ◽  
Germ Line ◽  
Sequence Similarity ◽  
Notch Receptor ◽  
Null Mutant ◽  
Reporter Protein ◽  
C Elegans

The C. elegans lag-2 gene is required for several cell-cell interactions that rely on the receptors GLP-1 and LIN-12. In this paper, we report that lag-2 encodes a putative membrane protein with sequence similarity to Drosophila Delta, a proposed ligand for the Notch receptor. Furthermore, we show that the lag-2 promoter drives expression of a reporter protein in the signaling distal tip cell (DTC) of the DTC/germline interaction. By in situ hybridization, we have found that endogenous lag-2 mRNA is present in the DTC but not the germ line. One fusion protein, called LAG-2::beta-gal(intra), rescues a lag-2 null mutant and can be detected in both DTC and germ line. Taking these results together, we propose that lag-2 may encode a signaling ligand for GLP-1/LIN-12 and that the entire LAG-2 protein may be taken up into the receiving cell during induction by GLP-1 and lateral signaling by LIN-12.

A gene with sequence similarity to Drosophila engrailed is expressed during the development of the neural tube and vertebrae in the mouse

Development ◽  
1988 ◽  
Vol 104 (2) ◽  
pp. 305-316 ◽  
Author(s):  
D. Davidson ◽  
E. Graham ◽  
C. Sime ◽  
R. Hill
Keyword(s):  
In Situ Hybridization ◽  
Neural Tube ◽  
Mouse Embryo ◽  
Sequence Similarity ◽  
Anterior Part ◽  
Limb Bud ◽  
Tissue Type ◽  
Restricted Domains ◽  
Restricted Region

The mouse genes En-1 and En-2 display sequence similarity, in and around the homeobox region, to the engrailed family in Drosophila. This paper describes their pattern of expression in the 12.5-day mouse embryo as determined by in situ hybridization. En-2 is expressed in a subset of cells expressing En-1. Both genes are expressed in the developing midbrain and its junction with the hindbrain. In addition, En-1 is expressed in the floor of the hindbrain, a restricted ventrolateral segment of the neural tube throughout the trunk and anterior part of the tail, the dermatome of tail somites, the centrum and costal processes in developing vertebrae, a restricted region of facial mesenchyme and the limb-bud ectoderm. Supplementary studies of 9.5-day and 10.5-day embryos showed that the same pattern of expression pertained in the neural tube, but that expression in the somites is at first confined to the dermatome and later found at a low level in restricted sclerotomal regions. Both genes are expressed in restricted domains which do not cross tissue-type boundaries. In several instances, however, boundaries of expression lie within morphologically undifferentiated tissue. These results suggest that En-1 and En-2 may be involved in the establishment or maintenance of the spatial integrity of specific domains within developing tissues.

GLP-1 is localized to the mitotic region of the C. elegans germ line

Development ◽  
1994 ◽  
Vol 120 (10) ◽  
pp. 2901-2911 ◽  
Author(s):  
S.L. Crittenden ◽  
E.R. Troemel ◽  
T.C. Evans ◽  
J. Kimble
Keyword(s):  
Membrane Protein ◽  
Germ Line ◽  
Integral Membrane Protein ◽  
Membrane Receptor ◽  
Down Regulation ◽  
Western Blots ◽  
Tip Cell ◽  
C Elegans ◽  
Cell Signal ◽  
Translational Level

In C. elegans, germline mitosis depends on induction by the somatic distal tip cell (DTC) and on activity of the glp-1 gene. Using antibodies to GLP-1 protein, we have examined GLP-1 on western blots and by immunocytochemistry. GLP-1 is tightly associated with membranes of mitotic germline cells, supporting its identification as an integral membrane protein. Furthermore, GLP-1 is localized within the germ line to the mitotic region, consistent with the model that GLP-1 acts as a membrane receptor for the distal tip cell signal. Unexpectedly, GLP-1 and the zone of mitosis extend further than the DTC processes. We present three models by which the DTC may influence GLP-1 activity and thereby determine the zone of mitosis. The spatial restriction of GLP-1 appears to be controlled at the translational level in hermaphrodites. We suggest that down-regulation of GLP-1 may be required to effect the transition from mitosis into meiosis.

scholarly journals Germ line-specific DNA sequences are present on all five micronuclear chromosomes in Tetrahymena thermophila.

1983 ◽  
Vol 3 (11) ◽  
pp. 1909-1919 ◽  
Author(s):  
K M Karrer
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Tetrahymena Thermophila ◽  
Germ Line ◽  
Plasmid Vector ◽  
Ciliated Protozoan ◽  
Specific Sequence ◽  
Individual Probe ◽  
Vector Pbr322

The development of the macronucleus from the zygotic micronucleus in the ciliated protozoan Tetrahymena spp. involves the elimination of specific DNA sequences (M. C. Yao and M. Gorovsky, Chromosoma 48:1-18 1974). The present study demonstrates that micronucleus-specific DNA is present on all five of the micronuclear chromosomes. Fragments of micronuclear DNA from Tetrahymena thermophila were cloned in the plasmid vector pBR322. A procedure was developed to examine the organization of the cloned sequences in micro- and macronuclear DNA without nick translating each individual probe. Twenty-three percent of randomly selected DNA sequences examined by this method were micronucleus (germ line) specific. They were all members of families of repeated sequences. Hybridization of six micronucleus-specific DNA sequences to micronuclear DNA from nullisomic strains of T. thermophila, which are lacking one or more pairs of chromosomes in the micronucleus, suggested that these sequences are present on several chromosomes. One micronucleus-specific sequence was shown by in situ hybridization to be present on all five of the micronuclear chromosomes.

scholarly journals Germ line-specific DNA sequences are present on all five micronuclear chromosomes in Tetrahymena thermophila

1983 ◽  
Vol 3 (11) ◽  
pp. 1909-1919
Author(s):  
K M Karrer
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Tetrahymena Thermophila ◽  
Germ Line ◽  
Plasmid Vector ◽  
Ciliated Protozoan ◽  
Specific Sequence ◽  
Individual Probe ◽  
Vector Pbr322

The development of the macronucleus from the zygotic micronucleus in the ciliated protozoan Tetrahymena spp. involves the elimination of specific DNA sequences (M. C. Yao and M. Gorovsky, Chromosoma 48:1-18 1974). The present study demonstrates that micronucleus-specific DNA is present on all five of the micronuclear chromosomes. Fragments of micronuclear DNA from Tetrahymena thermophila were cloned in the plasmid vector pBR322. A procedure was developed to examine the organization of the cloned sequences in micro- and macronuclear DNA without nick translating each individual probe. Twenty-three percent of randomly selected DNA sequences examined by this method were micronucleus (germ line) specific. They were all members of families of repeated sequences. Hybridization of six micronucleus-specific DNA sequences to micronuclear DNA from nullisomic strains of T. thermophila, which are lacking one or more pairs of chromosomes in the micronucleus, suggested that these sequences are present on several chromosomes. One micronucleus-specific sequence was shown by in situ hybridization to be present on all five of the micronuclear chromosomes.

scholarly journals A Fluorescence In situ Hybridization Screen for E26 Transformation–Specific Aberrations: Identification of DDX5-ETV4 Fusion Protein in Prostate Cancer

2008 ◽  
Vol 68 (18) ◽  
pp. 7629-7637 ◽  
Author(s):  
Bo Han ◽  
Rohit Mehra ◽  
Saravana M. Dhanasekaran ◽  
Jindan Yu ◽  
Anjana Menon ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
In Situ Hybridization ◽  
Fusion Protein ◽  
Fluorescence In Situ Hybridization

Bcl-2 Expression in Langerhans' Cell Histiocytosis

1998 ◽  
Vol 1 (3) ◽  
pp. 210-215 ◽  
Author(s):  
Van H. Savell ◽  
Todd Sherman ◽  
Richard H. Scheuermann ◽  
Abdul M. Siddiqui ◽  
Linda R. Margraf
Keyword(s):  
In Situ Hybridization ◽  
Langerhans Cells ◽  
Langerhans Cell Histiocytosis ◽  
Germ Line ◽  
Normal Skin ◽  
Protein Product ◽  
Langerhans Cell ◽  
Genomic Rearrangement ◽  
Southern Analysis

Langerhans' cell histiocytosis (LCH) is an abnormal accumulation of dendritic histiocytes of unknown pathogenesis. It has recently been shown to be a clonal process. Bcl-2 is a proto-oncogene whose protein product is known to inhibit apoptosis. The overexpression of bcl-2 has been demonstrated in a number of neoplasms, presumably prolonging the survival of the neoplastic cells. We examined the expression of bcl-2 in normal Langerhans' cells in the skin and in LCH by immunohistochemistry for protein and in situ hybridization for mRNA to see if it could be implicated in the pathogenesis of this disorder. Additionally, we performed Southern analysis to determine if genomic rearrangement of the bcl-2 gene occurs in cases of LCH. Bcl-2 was not detected in normal skin Langerhans' cells. Eleven of thirteen cases of LCH demonstrated bcl-2 protein expression in the cytoplasm of the Langerhans' cells by immunohistochemistry, while 12 of 13 cases had evidence of bcl-2 mRNA by in situ hybridization. Southern analysis revealed a germ-line configuration of the bcl-2 gene in the five cases studied. These findings suggest that bcl-2 expression is present and up-regulated in pathologic Langerhans' cells, however, this overexpression does not appear to be due to genomic rearrangement.

scholarly journals The Effects of Age and Reproduction on the Lipidome of Caenorhabditis elegans

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Qin-Li Wan ◽  
Zhong-Lin Yang ◽  
Xiao-Gang Zhou ◽  
Ai-Jun Ding ◽  
Yuan-Zhu Pu ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Caenorhabditis Elegans ◽  
Double Mutant ◽  
Germ Line ◽  
High Resolution Mass Spectrometry ◽  
Notch Receptor ◽  
C Elegans ◽  
Age Related ◽  
Lipid Species ◽  
Lipidome Analysis

Aging is a complex life process, and a unified view is that metabolism plays key roles in all biological processes. Here, we determined the lipidomic profile of Caenorhabditis elegans (C. elegans) using ultraperformance liquid chromatography high-resolution mass spectrometry (UPLC-HRMS). Using a nontargeted approach, we detected approximately 3000 species. Analysis of the lipid metabolic profiles at young adult and ten-day-old ages among wild-type N2, glp-1 defective mutant, and double mutant daf-16;glp-1 uncovered significant age-related differences in the total amount of phosphatidylcholines (PC), sphingomyelins (SM), ceramides (Cer), diglycerides (DG), and triglycerides (TG). In addition, the age-associated lipid profiles were characterized by ratio of polyunsaturated (PUFA) over monounsaturated (MUFA) lipid species. Lipid metabolism modulation plays an important role in reproduction-regulated aging; to identify the variations of lipid metabolites during germ line loss-induced longevity, we investigated the lipidomic profiles of long-lived glp-1/notch receptor mutants, which have reproductive deficiency when grown at nonpermissive temperature. The results showed that there was some age-related lipid variation, including TG 40:2, TG 40:1, and TG 41:1, which contributed to the long-life phenotype. The longevity of glp-1 mutant was daf-16-dependent; the lipidome analysis of daf-16;glp-1 double mutant revealed that the changes of some metabolites in the glp-1 mutant were daf-16-dependent, while other metabolites displayed more complex epistatic patterns. We first conducted a comprehensive lipidome analysis to provide novel insights into the relationships between longevity and lipid metabolism regulated by germ line signals in C. elegans.

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