Nucleocytoplasmic interactions in the mouse embryo

Development ◽  
1986 ◽  
Vol 97 (Supplement) ◽  
pp. 277-289
Author(s):  
James McGrath ◽  
Davor Solter
Keyword(s):  
Gene Transcription ◽  
Mouse Embryo ◽  
Developmental Stages ◽  
Female Parent ◽  
Nuclear Transplantation ◽  
Gene Products ◽  
Cytoplasmic Expression ◽  
Amphibian Embryos ◽  
Cellular Compartments ◽  
Nucleocytoplasmic Interactions

Fertilized mammalian ova consist of haploid genomes derived from both parents and cytoplasmic components inherited largely from the female parent. These three cellular compartments must successfully interact with each other and with their environment for development to proceed. These interactions require the transposition of nuclear and cytoplasmic products between cellular compartments with resultant alteration of gene transcription and the cytoplasmic expression of preformed or newly synthesized gene products. We have investigated nuclear/cytoplasmic interactions in the mouse embryo via the microsurgical transfer of nuclei and cytoplasm. Experiments have specifically examined the ability of nuclei from later developmental stages or from a different species to support development, volume relationships between nuclear and cytoplasmic compartments, and the nonequivalency of the maternal and paternal genomic contributions to development. The ability of egg cytoplasm to alter the function of a variety of embryonic and adult nuclei and the ability of these nuclei to support development has been extensively tested in nuclear transplantation investigations in amphibian embryos.

scholarly journals Expression Characteristics of the Transfer-RelatedkilB Gene Product of Streptomyces Plasmid pIJ101: Implications for the Plasmid Spread Function

2001 ◽  
Vol 183 (4) ◽  
pp. 1339-1345 ◽  
Author(s):  
Gregg S. Pettis ◽  
Naomi Ward ◽  
Kevin L. Schully
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Secondary Metabolism ◽  
Gene Product ◽  
Gene Transcription ◽  
Cellular Differentiation ◽  
Developmental Stages ◽  
Agar Medium ◽  
Spread Function ◽  
Aerial Hyphae

ABSTRACT Intermycelial transfer of Streptomyces plasmid pIJ101 occurs prior to cellular differentiation and is mediated by plasmid functions that are also required for production of zones of growth-inhibited recipient cells (i.e., pocks) that develop around individual donors during mating on agar medium. Several other pIJ101 functions, including that of the kilB gene, whose unregulated expression on pIJ101 is lethal, are required for normal pock size and so have been postulated to mediate intramycelial spread of the plasmid throughout recipient cells. Using antibodies raised against a KilB fusion protein expressed in Escherichia coli, native KilB protein was detected throughout development of pIJ101-containing Streptomyces lividans cells, with the concentration of KilB increasing dramatically and reaching a maximum during the final stages (i.e., sporulation and secondary metabolism) of cellular differentiation. Insertion of the kilB gene of pIJ101 into the S. lividans chromosome in cells lacking the pIJ101 KorB protein, which normally represses kilB gene transcription, resulted in elevated but still temporally increasing amounts of KilB. The increased expression or accumulation of the KilB spread protein throughout cellular differentiation of S. lividans, which leads to maximum KilB concentrations during developmental stages that occur far later than when intermycelial transfer of pIJ101 is mediated, supports the existence of a subsequent intramycelial component to the pIJ101 spread function. The results also suggest that intramycelial spread of pIJ101 molecules within the recipient extends beyond intercompartmental movements within the substrate mycelia and includes undetermined steps within the spore-yielding aerial hyphae as well.

Nuclear transplantation in the mouse embryo by microsurgery and cell fusion

Science ◽  
1983 ◽  
Vol 220 (4603) ◽  
pp. 1300-1302 ◽  
Author(s):  
J McGrath ◽  
D Solter
Keyword(s):  
Cell Fusion ◽  
Mouse Embryo ◽  
Nuclear Transplantation

Spatial- and temporal-restricted pattern for amelogenin gene expression during mouse molar tooth organogenesis

Development ◽  
1988 ◽  
Vol 104 (1) ◽  
pp. 77-85 ◽  
Author(s):  
M.L. Snead ◽  
W. Luo ◽  
E.C. Lau ◽  
H.C. Slavkin
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Gene Transcription ◽  
Developmental Stages ◽  
Three Dimensional ◽  
Molar Tooth ◽  
Specific Expression ◽  
Amelogenin Gene ◽  
Stage Of Development

Position- and time-restricted amelogenin gene transcription was analysed in developing tooth organs using in situ hybridization with asymmetric complementary RNA probes produced from a cDNA specific to the mouse 26 × 10(3) Mr amelogenin. In situ analysis was performed on developmentally staged fetal and neonatal mouse mandibular first (M1) and maxillary first (M1) molar tooth organs using serial sections and three-dimensional reconstruction. Amelogenin mRNA was first detected in a cluster of ameloblasts along one cusp of the M1 molar at the newborn stage of development. In subsequent developmental stages, amelogenin transcripts were detected within foci of ameloblasts lining each of the five cusps comprising the molar crown form. The number of amelogenin transcripts appeared to be position-dependent, being more abundant on one cusp surface while reduced along the opposite surface. Amelogenin gene transcription was found to be bilaterally symmetric between the developing right and left M1 molars, and complementary between the M1 and M1 developing molars; indicating position-restricted gene expression resulting in organ stereoisomerism. The application of in situ hybridization to forming tooth organ geometry provides a novel strategy to define epithelial-mesenchymal signal(s) which are believed to be responsible for organ morphogenesis, as well as for temporal- and spatial-restricted tissue-specific expression of enamel extracellular matrix.

Expression Pattern of Connexin Gene Products at the Early Developmental Stages of the Mouse Cardiovascular System

1997 ◽  
Vol 81 (3) ◽  
pp. 423-437 ◽  
Author(s):  
Bruno Delorme ◽  
Edgar Dahl ◽  
Thérèse Jarry-Guichard ◽  
Jean-Paul Briand ◽  
Klaus Willecke ◽  
...  
Keyword(s):  
Cardiovascular System ◽  
Expression Pattern ◽  
Developmental Stages ◽  
Gene Products ◽  
Connexin Gene ◽  
Early Developmental Stages ◽  
Early Developmental

scholarly journals PROTEINS IN NUCLEOCYTOPLASMIC INTERACTIONS

1968 ◽  
Vol 39 (2) ◽  
pp. 404-414 ◽  
Author(s):  
David Prescott ◽  
Lester Goldstein
Keyword(s):  
Binding Sites ◽  
Interphase Nucleus ◽  
Nuclear Protein ◽  
Protein A ◽  
Amoeba Proteus ◽  
Nuclear Proteins ◽  
Nuclear Transplantation ◽  
Amino Acid Labeling ◽  
Turn Over ◽  
Nucleocytoplasmic Interactions

The behavior of nuclear proteins in Amoeba proteus was studied by tritiated amino acid labeling, nuclear transplantation, and cytoplasmic amputation. During prophase at least 77% (but probably over 95%) of the nuclear proteins is released to the cytoplasm. These same proteins return to the nucleus within the first 3 hr of interphase. When cytoplasm is amputated from an ameba in mitosis (shen the nuclear proteins are in the cytoplasm), the resultant daughter nuclei are depleted in the labeled nuclear proteins. The degree of depletion is less than proportional to the amount of cytoplasm removed because a portion of rapidly migrating protein (a nuclear protein that is normally shuttling between nucleus and cytoplasm and is thus also present in the cytoplasm) which would normally remain in the cytoplasm is taken up by the reconstituting daughter nuclei. Cytoplasmic fragments cut from mitotic cells are enriched in both major classes of nuclear proteins, i.e. rapidly migrating protein and slow turn-over protein. An interphase nucleus implanted into such an enucleated cell acquires from the cytoplasm essentially all of the excess nuclear proteins of both classes. The data indicate that there is a lack of binding sites in the cytoplasm for the rapidly migrating nuclear protein. The quantitative aspects of the distribution of rapidly migrating protein between the nucleus and the cytoplasm indicate that the distribution is governed primarily by factors within the nucleus.

scholarly journals An integrated genome-wide multi-omics analysis of gene expression dynamics in the preimplantation mouse embryo

2018 ◽  
Author(s):  
Steffen Israel ◽  
Mathias Ernst ◽  
Olympia E. Psathaki ◽  
Hannes C. A. Drexler ◽  
Ellen Casser ◽  
...  
Keyword(s):  
Mouse Embryo ◽  
Developmental Stages ◽  
Protein Complexes ◽  
Large Fraction ◽  
Transcript Level ◽  
Free Ribosome ◽  
Translational Machinery ◽  
Preimplantation Mouse ◽  
Morula Stage ◽  
The Impact

AbstractEarly mouse embryos have an atypical translational machinery comprised of cytoplasmic lattices, poorly competent for translation. Thus, the impact of transcriptomic changes on the operational levels of proteins has likely been overestimated in the past. To find out, we used liquid chromatography–tandem mass spectrometry to detect and quantify 6,550 proteins in the oocyte and in six developmental stages (from zygote to blastocyst) collected in triplicates, and we also performed mRNA sequencing.In contrast to the known split between the 2-cell and 4-cell stages at the transcript level, on the protein level the oocyte-to-embryo transition appeared to last until the morula stage. In general, protein abundance profiles were weakly correlated with those of their cognate mRNAs and we found little or no concordance between changes in protein and transcript expression relative to the oocyte at early stages. However, concordance increased towards morula and blastocyst, hinting at a more direct coupling of proteins with transcripts at these stages, in agreement with the increase in free ribosome abundance. Independent validation by immunofluorescence and qPCR confirmed the existence of genes featuring strongly positively and negatively correlated protein and transcript. Moreover, consistent coverage of most known protein complexes indicates that our dataset represents a large fraction of the expressed proteome. Finally, we identified 20 markers, including members of the endoplasmic reticulum pathway, for discriminating between early and late stages.This resource contributes towards closing the gap between the ‘predicted’ phenotype, based on mRNA, and the ‘actual’ phenotype, based on protein, of the mouse embryo.

scholarly journals Low Quantity single strand CAGE (LQ-ssCAGE) maps regulatory enhancers and promoters

2020 ◽  
Author(s):  
Hazuki Takahashi ◽  
Hiromi Nishiyori-Sueki ◽  
Jordan A. Ramilowski ◽  
Masayoshi Itoh ◽  
Piero Carninci
Keyword(s):  
Gene Expression ◽  
Developmental Stages ◽  
Cost Effective ◽  
Single Strand ◽  
Large Cage ◽  
Transcription Start Sites ◽  
Dna Elements ◽  
Nucleotide Resolution ◽  
Cap Analysis ◽  
Cellular Compartments

AbstractThe Cap Analysis of Gene Expression (CAGE) is a powerful method to identify the Transcription Start Sites (TSSs) of capped RNAs while simultaneously measuring transcripts expression level. CAGE allows mapping at single nucleotide resolution at all active promoters and enhancers. Large CAGE datasets have been produced over the years from individual laboratories and consortia, including the Encyclopedia of DNA Elements (ENCODE) and Functional Annotation Of the Mammalian Genome (FANTOM). These datasets constitute open resource for TSS annotations and gene expression analysis. Here we provide an experimental protocol for the most recent CAGE method called Low Quantity (LQ) single strand (ss) CAGE “LQ-ssCAGE”, which enables cost-effective profiling of low quantity RNA samples. LQ-ssCAGE is especially useful for samples derived from cells cultured in small volumes, cellular compartments such as nuclear RNAs or for samples from developmental stages. We demonstrate the reproducibility and effectiveness of the method by constructing 240 LQ-ssCAGE libraries from 50 ng of THP-1 cell extracted RNAs and discover lowly expressed novel enhancer and promoter-derived lncRNAs.

Isolation and analysis of a novel class of suppressor of Ty insertion mutations in Saccharomyces cerevisiae.

Genetics ◽  
1988 ◽  
Vol 118 (2) ◽  
pp. 203-212
Author(s):  
J S Fassler ◽  
F Winston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Transcription ◽  
Transcriptional Analysis ◽  
Insertion Mutation ◽  
Gene Products ◽  
Adjacent Gene ◽  
Cellular Functions ◽  
Ty Elements ◽  
Insertion Mutations

Abstract Using a new scheme for the isolation of suppressor of Ty insertion mutations (spt mutations) in yeast, we have identified six new SPT genes. Mutations in two of these genes, SPT13 and SPT14, exhibit a novel suppression pattern: suppression of complete Ty insertion mutations, but not of solo delta insertion mutations. Transcriptional analysis shows that spt13- and spt14-mediated suppression of Ty insertion mutations is the result of an elevation in the levels of adjacent gene transcription. In spite of the failure of these mutations to suppress solo delta insertion mutations, they do cause changes in transcription of at least one solo delta insertion mutation. In addition, spt13 and spt14 mutations are epistatic to mutations in certain other SPT genes that do suppress solo delta insertion mutations. These results suggest that the SPT13 and SPT14 gene products may act via sequences in both the delta and epsilon regions of Ty elements. Finally, mutations in SPT13 cause sporulation and mating defects and SPT14 is essential for growth, suggesting that these two genes have important roles in general cellular functions.

scholarly journals Insulating DNA directs ubiquitous transcription of the Drosophila melanogaster alpha 1-tubulin gene.

1994 ◽  
Vol 14 (9) ◽  
pp. 6398-6408 ◽  
Author(s):  
K H O'Donnell ◽  
C T Chen ◽  
P C Wensink
Keyword(s):  
Drosophila Melanogaster ◽  
Developmental Stages ◽  
Germ Line ◽  
Core Promoter ◽  
High Sensitivity ◽  
In Vitro Transcription ◽  
Gene Products ◽  
Position Effects ◽  
Germ Line Transformation

We identify DNA regions that are necessary for the ubiquitous expression of the Drosophila melanogaster alpha 1-tubulin (alpha 1t) gene. In vitro transcription showed that two upstream regions, tubulin element 1 (TE1 [29 bp]) and tubulin element 2 (TE2 [68 bp]), and a downstream region activate transcription. Germ line transformation demonstrated that these three regions are sufficient to direct the alpha 1t core promoter to begin transcribing at the stage of cellular blastoderm formation and to continue thereafter at high levels in all tissues and developmental stages. Remarkably, mutation of any one of these regions results in high sensitivity to chromosomal position effects, producing different but reproducible tissue-specific patterns of expression in each transformed line. None of these regions behaves as an enhancer in a conventional germ line transformation test. These observations show that these three regions, two of which bind the GAGA transcription factor, act ubiquitously to insulate from position effects and to activate transcription. The results also provide vectors for ubiquitous expression of gene products and for examining silencer activities.

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