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At least 883 records · Page 49Linked to original sources
PubMed2001

Mammalian myotube dedifferentiation induced by newt regeneration extract.

Newts are capable of regenerating several anatomical structures and organs, including their limbs. This remarkable regenerative capacity is thought to depend on cellular dedifferentiation. Terminally differentiated mammalian cells, by contrast, are normally incapable of reversing the differentiation process. Several factors could explain the absence of cellular dedifferentiation in mammals: (i) inadequate expression of genes that initiate dedifferentiation; (ii) insufficient intracellular signaling pathways; (iii) irreversible expression of differentiation factors; and (iv) structural characteristics that make dedifferentiation impossible. To investigate the causes underlying the lack of cellular plasticity in mammalian cells, we examined the effect of an extract derived from newt regenerating limbs on terminally differentiated mouse C2C12 myotubes. Approximately 18% of murine myotubes reentered the cell cycle when treated with regeneration extract, whereas 25% of newt myotubes exhibited cell cycle reentry. The muscle differentiation proteins MyoD, myogenin, and troponin T were reduced to undetectable levels in 15-30% of treated murine myotubes. We observed cellular cleavage in 11% of the treated murine myotubes and approximately 50% of these myotubes continued to cleave to produce proliferating mononucleated cells. These data indicate that mammalian myotubes can dedifferentiate when stimulated with the appropriate factors and suggest that one mechanism preventing dedifferentiation of mammalian cells is inadequate spatial or temporal expression of genes that initiate dedifferentiation.

Animals↗
PubMed2025

Profiling hippocampal neuronal populations reveals unique gene expression mosaics reflective of connectivity-based degeneration in the Ts65Dn mouse model of Down syndrome and Alzheimer's disease.

INTRODUCTION: Individuals with Down syndrome (DS) exhibit neurological deficits throughout life including the development of in Alzheimer's disease (AD) pathology and cognitive impairment. At the cellular level, dysregulation in neuronal gene expression is observed in postmortem human brain and mouse models of DS/AD. To date, RNA-sequencing (RNA-seq) analysis of hippocampal neuronal gene expression including the characterization of discrete circuit-based connectivity in DS remains a major knowledge gap. We postulate that spatially characterized hippocampal neurons display unique gene expression patterns due, in part, to dysfunction of the integrity of intrinsic circuitry. METHODS: We combined laser capture microdissection to microisolate individual neuron populations with single population RNA-seq analysis to determine gene expression analysis of CA1 and CA3 pyramidal neurons and dentate gyrus granule cells located in the hippocampus, a region critical for learning, memory, and synaptic activity. RESULTS: The hippocampus exhibits age-dependent neurodegeneration beginning at ~6 months of age in the Ts65Dn mouse model of DS/AD. Each population of excitatory hippocampal neurons exhibited unique gene expression alterations in Ts65Dn mice. Bioinformatic inquiry revealed unique vulnerabilities and differences with mechanistic implications coinciding with onset of degeneration in this model of DS/AD. CONCLUSIONS: These cell-type specific vulnerabilities may underlie degenerative endophenotypes suggesting precision medicine targeting of individual populations of neurons for rational therapeutic development.

Alzheimer’s disease↗
PubMed1993

The process of reinnervation in the dentate gyrus of adult rats: temporal relationship between changes in the levels of glial fibrillary acidic protein (GFAP) and GFAP mRNA in reactive astrocytes.

The present study evaluates the temporal relationships between increases in glial fibrillary acidic protein (GFAP) mRNA, GFA protein levels, and GFAP immunostaining in the hippocampus of adult rats following unilateral lesions of the entorhinal cortex (EC). GFAP mRNA levels were assessed at 12 h, 1, 2, 4, 6, 8, 10, 12, 14, and 30 days postlesion by dot blot assays using 35S-labeled cRNA probes against the mRNA. Animals were also prepared for in situ hybridization during the peak of GFAP mRNA expression (2 days postlesion) to explore the nature of individual differences in the spatial extent of the increases. GFA protein levels were assessed by Western blot and dot immunoblot techniques in a separate group of animals prepared at 1, 2, 4, 6, 8, and 10 days postlesion and by immunostaining at 1, 2, 4, 6, and 8 days postlesion. The dot blot analyses of GFAP mRNA levels confirmed previous studies, in that we observed dramatic increases in the levels of GFAP mRNA in the hippocampus ipsilateral to the EC lesions. The increases were biphasic, with a large peak in mRNA levels at 1-2 days postlesion (about 10-fold greater than control) and a second peak at 6-8 days. In most animals, the increases were predominantly ipsilateral to the lesion. However, in some animals, there were also large increases on the contralateral side. In situ hybridization experiments revealed two different spatial patterns of increased gene expression, one in which the increases in GFAP mRNA occurred bilaterally and one in which increases were restricted primarily to the hippocampus ipsilateral to the lesion. Immunochemical measures revealed that GFA protein levels increased gradually in the hippocampus ipsilateral to the lesion, reaching a peak at about 2-fold higher than control at 4 days postlesion, and then remained near this level until at least 10 days postlesion. In the contralateral hippocampus, GFA protein levels were increased to about the same extent as on the ipsilateral side at 1, 2, and 4 days postlesion, but then began to decline, returning to near control levels by 8 days. Increases in immunostaining occurred with about the same time course as the increases in GFA protein levels as measured immunochemically. These results define the temporal relationship between increases in GFAP mRNA and increases in GFA protein, providing new insights into the regulation of gene expression in reactive astrocytes.

Animals↗
PubMed2001

Erythropoietin (Epo) and EpoR expression and 2 waves of erythropoiesis.

Erythropoiesis occurs in 2 distinct waves during embryogenesis: the primitive wave in the extra-embryonic yolk sac (YS) followed by the definitive wave in the fetal liver and spleen. Even though progenitors for both cell types are present in the YS blood islands, only primitive cells are formed in the YS during early embryogenesis. In this study, it is proposed that erythropoietin (Epo) expression and the resultant EpoR activation regulate the timing of the definitive wave. First, it was demonstrated that Epo and EpoR gene expressions are temporally and spatially segregated: though EpoR is expressed early (embryonic days 8.0-9.5) in the yolk sac blood islands, no Epo expression can be detected in this extra-embryonic tissue. Only at a later stage can Epo expression be detected intra-embryonically, and the onset of Epo expression correlates with the initiation of definitive erythropoiesis. It was further demonstrated that the activation of the EpoR signaling pathway by knocking-in a constitutively active form of EpoR (R129C EpoR) can lead to earlier onset of definitive erythropoiesis in the YS. Thus, these results provide the first in vivo mechanism as to how 2 erythroid progenitor populations can coexist concurrently in the YS yet always differentiate successively during embryogenesis.

Animals↗
PubMed2003

Subnuclear localization of C/EBP beta is regulated by growth hormone and dependent on MAPK.

Localization of transcription regulatory proteins in the nucleus is dynamically regulated, and may alter nucleoplasmic concentrations and/or assembly of multimolecular transcription regulatory complexes, which ultimately regulate gene expression. Since growth hormone (GH) regulates multiple transcription factors including C/EBP beta, the effect of GH on the subcellular localization of C/EBP beta was examined in 3T3-F442A preadipocytes. Indirect immunofluorescence shows that C/EBP beta is diffusely distributed in nuclei of quiescent cells. Within 5 min of GH treatment, the diffuse pattern dramatically becomes punctate. The relocalization of C/EBP beta coincides with DAPI staining of heterochromatin. Further, C/EBP beta and heterochromatin protein (HP)-1 alpha colocalize in the nucleus, consistent with localization of C/EBP beta to pericentromeric heterochromatin. In contrast, C/EBP delta exhibits a diffuse distribution in the nucleus that is not modified by GH treatment. C/EBP beta is rapidly and transiently phosphorylated on a conserved MAPK consensus site in response to GH (Piwien-Pilipuk, G., MacDougald, O. A., and Schwartz, J. (2002) J. Biol. Chem. 277, 44557-44565). Indirect immunofluorescence using antibodies specific for C/EBP beta phosphorylated on the conserved MAPK site shows that GH also rapidly induces a punctate pattern of staining for the phosphorylated C/EBP beta. In addition, phosphorylated C/EBP beta colocalizes to pericentromeric heterochromatin. The satellite DNA present in heterochromatin contains multiple C/EBP binding sites. DNA binding analysis shows that C/EBP beta, C/EBP delta, and C/EBP alpha (p42 and p30 forms) can bind to satellite DNA as homo- or heterocomplexes in vitro. Importantly, GH rapidly and transiently increases binding of endogenous C/EBP beta from 3T3-F442A cells to satellite DNA. Further, the GH-promoted nuclear relocalization of C/EBP beta to pericentromeric heterochromatin was prevented by the MEK inhibitor U0126. This observation suggests that GH-dependent MAPK activation plays a role in the regulation of nuclear relocalization of C/EBP beta. Nuclear redistribution introduces a new level of transcriptional regulation in GH action, since GH-mediated phosphorylation and nuclear redistribution of C/EBP beta may be coordinated to achieve spatial-temporal control of gene expression.

3T3 Cells↗
PubMed1995

Temporal expression and cellular distribution of pulmonary fibronectin gene induction following exposure to an emission source particle.

This study examines the ability of an emission source particle, residual oil fly ash (ROFA), to influence pulmonary fibronectin (Fn) gene expression. Fn is an extracellular matrix (ECM) protein involved in a variety of cellular functions, including inflammation, cell proliferation, and fibrosis. Temporal expression and spatial distribution of Fn gene induction were assessed by in situ hybridization in rat lung during acute phase of lung injury occurring 6 to 72 h following intratracheal instillation of ROFA. Fn mRNA was not detected in rat lungs treated with either saline or at 6 h after ROFA treatment. However, Fn mRNA was induced in airway epithelial cells 24 h after ROFA instillation. Histopathology showed peribronchial inflammation and focal edema. Diffuse inflammation in alveolar region with limited expression of Fn mRNA was evident 48 h after ROFA exposure, occurring mainly in proliferating epithelial cells. Extensive Fn mRNA expression was seen in proliferating fibroblasts and in hyperplastic epithelial cells within incipient fibrotic lesions 72 h after exposure, while the intensity of expression in the airway epithelial cells was decreased. Therefore, Fn mRNA induction was associated with inflammatory and incipient fibrotic lesions, indicating its possible involvement in airway hyperreactivity and initiation of fibrogenesis.

Air Pollutants↗
PubMed1991

A yeast transcription assay defines distinct rel and dorsal DNA recognition sequences.

Recent data have demonstrated that vRel, cRel, Dorsal, and NF-kappa B are members of a larger family of DNA-binding regulatory proteins. Rel proteins interact to form homo- and heterodimers that recognize specific sites on DNA, and it is likely that such protein-protein and protein-DNA interactions contribute to proper regulation of target gene expression by these proteins. Here we describe the use of a yeast transcription activation assay to study binding of three Rel family proteins to their native binding sites. These results show that the vRel and cRel proteins recognize two known NF-kappa B binding sites; the Dorsal protein does not recognize NF-kappa B sites, but does recognize related sites upstream of the Drosophila zerknüllt gene. Our experiments demonstrate that the members of this protein family recognize similar, but not identical, sites in the promoters of target genes, and we are able to identify a particular nucleotide that is apparently involved in the DNA-protein interaction. We exploit the properties of LexA fusion proteins to study the dimerization and DNA-contacting domains of cRel. Our results suggest that the cRel protein forms homodimers and that dimer formation may be necessary for cRel to bind DNA. Finally, our results show that transcription activation by these proteins is cooperative; such cooperativity may be important for correct temporal and spatial regulation of target gene expression.

Animals↗
PubMed2003

Coupling gene expression and multicellular morphogenesis during fruiting body formation in Myxococcus xanthus.

A recurring theme in morphogenesis is the coupling of the expression of genes that drive morphogenesis and the morphogenetic process per se. This coupling ensures that gene expression and morphogenesis are carried out in synchrony. Morphogenesis of the spore-filled fruiting bodies in Myxococcus xanthus illustrates this coupling in the construction of a multicellular structure. Fruiting body formation involves two stages: aggregation of cells into mounds and the position-specific sporulation of cells that have accumulated inside mounds. Developmental gene expression propels these two processes. In addition, gene expression in individual cells is adjusted according to their spatial position. Progress in the understanding of the cell surface-associated C-signal is beginning to reveal the framework of an intercellular signalling system that allows the coupling of gene expression and multicellular morphogenesis. Accumulation of the C-signal is tightly regulated and involves transcriptional activation of the csgA gene and proteolysis of the full-length CsgA protein to produce the shorter cell surface-associated 17 kDa C-signal protein. The C-signal induces aggregation, sporulation and developmental gene expression at specific thresholds. The ordered increase in C-signalling levels, in combination with the specific thresholds, allows the C-signal to induce these three processes in the correct temporal order. The contact-dependent C-signal transmission mechanism, in turn, guarantees that C-signalling levels reflect the spatial position of individual cells relative to other cells and, thus, allows the cells to decode their spatial position during morphogenesis. By this mechanism, individual cells can tailor their gene expression profile to one that matches their spatial position. In this scheme, the molecular device that keeps gene expression in individual cells in register with morphogenesis is the C-signalling system, and the morphological structure, which is assessed, is the spatial position of individual cells relative to that of other cells.

Gene Expression Regulation, Bacterial↗
PubMed1999

Down-regulation of L-type calcium channel and sarcoplasmic reticular Ca(2+)-ATPase mRNA in human atrial fibrillation without significant change in the mRNA of ryanodine receptor, calsequestrin and phospholamban: an insight into the mechanism of atrial electrical remodeling.

OBJECTIVES: We investigated the gene expression of calcium-handling genes including L-type calcium channel, sarcoplasmic reticular calcium adenosine triphosphatase (Ca(2+)-ATPase), ryanodine receptor, calsequestrin and phospholamban in human atrial fibrillation. BACKGROUND: Recent studies have demonstrated that atrial electrical remodeling in atrial fibrillation is associated with intracellular calcium overload. However, the changes of calcium-handling proteins remain unclear. METHODS: A total of 34 patients undergoing open heart surgery were included. Atrial tissue was obtained from the right atrial free wall, right atrial appendage, left atrial free wall and left atrial appendage, respectively. The messenger ribonucleic acid (mRNA) amount of the genes was measured by reverse transcription-polymerase chain reaction and normalized to the mRNA levels of glyceraldehyde 3-phosphate dehydrogenase. RESULTS: The mRNA of L-type calcium channel and of Ca(2+)-ATPase was significantly decreased in patients with persistent atrial fibrillation for more than 3 months (0.36+/-0.26 vs. 0.90+/-0.88 for L-type calcium channel; 0.69+/-0.42 vs. 1.21+/-0.68 for Ca(2+)-ATPase; both p < 0.05, all data in arbitrary unit). We further demonstrated that there was no spatial dispersion of the gene expression among the four atrial tissue sampling sites. Age, gender and underlying cardiac disease had no significant effects on the gene expression. In contrast, the mRNA levels of ryanodine receptor, calsequestrin and phospholamban showed no significant change in atrial fibrillation. CONCLUSIONS: L-type calcium channel and the sarcoplasmic reticular Ca(2+)-ATPase gene were down-regulated in atrial fibrillation. These changes may be a consequence of, as well as a contributory factor for, atrial fibrillation.

Adolescent↗
PubMed2002

Activation of Rac1 by shear stress in endothelial cells mediates both cytoskeletal reorganization and effects on gene expression.

Hemodynamic shear stress is a fundamental determinant of vascular remodeling and atherogenesis. Changes in focal adhesions, cytoskeletal organization and gene expression are major responses of endothelial cells to shear stress. Here, we show that activation of the small GTPase Rac is essential for gene expression and for providing spatial information for shear stress-induced cell alignment. Fluorescence resonance energy transfer (FRET) localizes activated Rac1 in the direction of flow. This directional Rac1 activation is downstream of shear-induced new integrin binding to extracellular matrix. Additionally, Rac1 mediates flow-induced stimulation of nuclear factor kappaB (NF-kappaB) and the subsequent expression of intercellular cell adhesion molecule 1 (ICAM-1), an adhesion receptor involved in the recruitment of leukocytes to atherosclerotic plaque. These studies provide a unifying model linking three of the main responses to shear stress that mediate both normal adaptation to hemodynamic forces and inflammatory dysfunction of endothelial cells in atherosclerosis.

Animals↗
PubMed2006

Retinoids, eye development, and maturation of visual function.

Vitamin A is known to be critical for the beginning of eye development as well as for photoreception in the functional retina. Hardly anything, however, is known about whether retinoic acid (RA)-regulated gene expression also plays a role in the long intervening period, during which the neurobiological retinal structure takes shape. The eye contains a highly intricate architecture of RA-synthesizing (RALDH) and degrading (CYP26) enzymes. Whereas the RALDHs are integrated in the early molecular mechanisms through which the dorso-ventral retina organization is established, the CYP26 enzymes are not necessary for this process and no molecular targets that match their retinal expression pattern have yet been identified. In this article we describe that CYP26 expression in the mouse is most distinctive during later stages of retina formation. Throughout development CYP26A1 degrades RA in a horizontal region that extends across the retina, but during later embryonic and postnatal retina maturation this function is reinforced by another enzyme, CYP26C1. RA applications at this stage do not affect the RALDHs but cause differential changes in CYP26 expression: Cyp26a1 is up-regulated, but more rapidly by 9-cis than all-trans RA, Cyp26c1 is down-regulated, and Cyp26b1, which is undetectable in the normal mouse retina, is strongly activated in retinal ganglion cells. The dynamic regulation in RA-difference patterns by the CYP26 enzymes may set up spatial constellations for expression of genes involved in formation of retinal specializations for higher acuity vision, which are known to form over a prolonged period late in retina development.

Aldehyde Oxidoreductases↗
PubMed1994

Genomic organization and chromosome localization of the murine homeobox gene Pmx.

Homeobox genes are expressed in very specific temporal and spatial patterns and function as transcriptional regulators of developmental processes. The murine homeobox gene, Pmx (paired mesoderm homeobox), is expressed in a mesodermally restricted pattern in embryos and most abundantly in cardiac, skeletal, and smooth muscle tissues in adults. Previously, this murine gene was named K-2 and mHox, while the human homolog was named Phox1. In this report, the localization of Pmx has been determined by interspecific backcross analysis. The Pmx gene is located on Chromosome 1, approximately 3.3 cM distal to the Gsh-4 homeobox locus. The sequence of the Pmx transcript has been extended toward the 5' end and corresponds in size to one of the transcripts previously detected by Northern blot analysis. Sequence analysis indicates that Pmx is the first characterized mammalian gene to encode a paired type homeodomain, but not a paired domain. The Pmx gene includes at least five exons spanning a minimum of 60 kb of genomic DNA, making this the largest known murine homeobox gene.

Animals↗
PubMed2005

Replication, variation and normalisation in microarray experiments.

INTRODUCTION: Microarray experiments often have complex designs that include sample pooling, biological and technical replication, sample pairing and dye-swapping. This article demonstrates how statistical modelling can illuminate issues in the design and analysis of microarray experiments, and this information can then be used to plan effective studies. METHODS: A very detailed statistical model for microarray data is introduced, to show the possible sources of variation that are present in even the simplest microarray experiments. Based on this model, the efficacy of common experimental designs, normalisation methodologies and analyses is determined. RESULTS: When the cost of the arrays is high compared with the cost of samples, sample pooling and spot replication are shown to be efficient variance reduction methods, whereas technical replication of whole arrays is demonstrated to be very inefficient. Dye-swap designs can use biological replicates rather than technical replicates to improve efficiency and simplify analysis. When the cost of samples is high and technical variation is a major portion of the error, technical replication can be cost effective. Normalisation by centreing on a small number of spots may reduce array effects, but can introduce considerable variation in the results. Centreing using the bulk of spots on the array is less variable. Similarly, normalisation methods based on regression methods can introduce variability. Except for normalisation methods based on spiking controls, all normalisation requires that most genes do not differentially express. Methods based on spatial location and/or intensity also require that the nondifferentially expressing genes are at random with respect to location and intensity. Spotting designs should be carefully done so that spot replicates are widely spaced on the array, and genes with similar expression patterns are not clustered. DISCUSSION: The tools for statistical design of experiments can be applied to microarray experiments to improve both efficiency and validity of the studies. Given the high cost of microarray experiments, the benefits of statistical input prior to running the experiment cannot be over-emphasised.

Algorithms↗
PubMed2005

Posttraining intrahippocampal infusion of a protein kinase AII inhibitor impairs spatial memory retention in rats.

The role of protein kinase AII (PKA II) in spatial memory retention in male rats and its regulation of cholinergic gene expression were explored through the effects of intrahippocampal infusion of H-89, a selective PKA II inhibitor. Alterations in escape latency, travel distance, and swimming speed in a Morris water maze were measured. Animals were trained for 3 days; each day included two blocks, and each block contained four trials. Stereotaxic surgery was employed for the infusions after the last trial on the third day of training, and the animals were tested 48 hr after surgery. Bilateral intrahippocampal infusion of H-89 (2.5 or 5 microM) into the CA1 region generated significant alterations in escape latency and traveled distance but not swimming speed. The response was fairly dose dependent, and the maximal effect was obtained with 5 microM H-89. After behavioral testing, several of the infused animals were transcardially perfused and their brains removed. Brain tissue sections from these rats were subjected to immunohistochemical staining analysis with anticholine acetyltransferase (ChAT) antibodies. These analyses indicated that 5 microM H-89 infusions qualitatively reduced the density of ChAT-containing cholinergic nerve terminals in the dorsal hippocampus. The intrahippocampal infusions with 5 microM H-89 also caused an apparent reduction in the number of ChAT-containing neurons in the medial septum. Our results suggest that PKA II is involved in regulation of cholinergic gene expression and plays an important role in spatial memory retention in rats.

Acetylcholine↗
PubMed1998

Sugars and light/dark exposure trigger differential regulation of ADP-glucose pyrophosphorylase genes in Arabidopsis thaliana (thale cress).

Expression of four Arabidopsis (thale cress) genes corresponding to the small (ApS) and large subunits (ApL1, ApL2, ApL3) of ADP-glucose pyrophosphorylase (AGPase), a key enzyme of starch biosynthesis, was found to be profoundly and differentially regulated by sugar and light/dark exposures. Transcript levels of both ApL2 and ApL3, and to a lesser extent ApS, increased severalfold upon feeding sucrose or glucose to the detached leaves in the dark, whereas the mRNA content for ApL1 decreased under the same conditions. Glucose was, in general, less effective than sucrose in inducing regulation of AGPase genes, possibly due to observed limitations in its uptake when compared with sucrose uptake by detached leaves. Osmotic agents [sorbitol, poly(ethylene glycol)] had no effect on ApS, ApL2 and ApL3 transcript level, but they did mimic the effect of sucrose on ApL1 gene, suggesting that the latter is regulated by osmotic pressure rather than any particular sugar. For all the genes the sugar effect was closely mimicked by an exposure of the dark-pre-adapted leaves to the light. Under both dark and light conditions, sucrose fed to the detached leaves was found to be rapidly metabolized to hexoses and, to some extent, starch. Starch production reflected most probably an increase in substrate availability for AGPase reaction rather than being due to changes in AGPase protein content, since both the sugar feeding and light exposure had little or no effect on the activity of AGPase or on the levels of its small and large subunit proteins in leaf extracts. The data suggest tight translational or post-translational control, but they may also reflect spatial control of AGPase gene expression within a leaf. The sugar/light-dependent regulation of AGPase gene expression may represent a part of a general cellular response to the availability/allocation of carbohydrates during photosynthesis.

Arabidopsis↗
PubMed1997

Molecular organization of a gene in barley which encodes a protein similar to aspartic protease and its specific expression in nucellar cells during degeneration.

The nucellar cells of barley undergo progressive degeneration after ovule fertilization. This degeneration is a characteristic of programmed cell death. Increasing evidence has indicated that proteases are important regulators of programmed cell death in animals. We have cloned and characterized a barley gene which encodes an aspartic protease-like protein and is specifically expressed in nucellar cells during degeneration. The gene contains eight exons and seven introns and encodes a polypeptide of 410 amino acid residues. The deduced polypeptide is characterized by having two aspartic protease catalytic site motifs, the Asp-Thr-Gly-Ser in the N-terminal and Asp-Ser-Gly-Ser in the C-terminal region, and two other regions nearly identical to two regions of plant aspartic proteases. However, it shares < 20% overall sequence identity with the known plant aspartic proteases, and does not contain a 'prosequence' or a 'plant-specific insert' which are characteristics of plant aspartic proteases. We have named this aspartic protease-like protein 'nucellin'. In northern analyses nucellin transcripts were most abundant in ovaries 3-4 days after pollination, but only marginally detectable before pollination or 10 days after pollination. RNA in situ hybridization showed that before pollination the nucellin gene was expressed at a very low level only in a cluster of nucellar cells close to the embryo sac at the chalazal end, but after pollination it was highly expressed in most nucellar cells surrounding the entire embryo sac. Furthermore, no nucellin transcripts were detectable in anther, leaf, or root tissue. The temporal and spatial pattern of the nucellin gene expression is synchronal with nucellar cell degeneration and thus, nucellin may be involved with nucellar cell death.

Amino Acid Sequence↗
PubMed1991

The muscle regulatory gene, Myf-6, has a biphasic pattern of expression during early mouse development.

The spatial and temporal expression pattern of the muscle regulatory gene Myf-6 (MRF4/herculin) has been investigated by in situ hybridization during embryonic and fetal mouse development. Here, we report that the Myf-6 gene shows a biphasic pattern of expression. Myf-6 transcripts are first detected in the most rostral somites of the mouse embryo at 9 d of gestation and accumulate progressively in myotomal cells along the rostro-caudal axis. This expression is transient and Myf-6 mRNA can no longer be detected in myotomal cells after day 12 post coitum (p.c.). In contrast to other muscle determination genes (MyoD1, myogenin, Myf-5), Myf-6 mRNA is not detected in limb buds or visceral arches and skeletal muscle of the mouse embryo (day 8-15 p.c.). In fetal mice, Myf-6 transcripts appear at day 16 p.c. in all skeletal muscles, and the gene continues to be expressed at a high level after birth. These results suggest that early Myf-6 expression may be restricted to a population of myogenic cells that does not contribute to the embryonic muscle masses in limb buds and visceral arches. The reappearance of Myf-6 mRNA in fetal skeletal muscle coincides approximately with secondary muscle fiber formation and the onset of innervation.

Base Sequence↗
PubMed2000

Photoactivated gene expression for cell fate mapping and cell manipulation.

A long-standing goal of developmental biologists is to create developmental fate maps by tracking individual cells through development. Another objective is to perturb the behavior of selected cells and follow the ensuing effects. To this end, we have developed a technique that allows for spatial and temporal control of gene expression in single cells or patches of cells using light to induce gene expression. This technique relies on "caging" the activity of the potent transcriptional activator GAL4VP16 with a photolabile compound, which can be removed with a brief exposure to long-wavelength ultraviolet (UV) light. The caged GAL4VP16 is injected into early-stage embryos, which are aged to the desired point in development, and the cell(s) of interest are irradiated with a brief pulse of long-wavelength UV light. This method has been used extensively in Drosophila, Xenopus, and Zebrafish embryos. The methods for purifying, caging, injection, and photoactivation of the GAL4VP16 protein, and methods for the visualization of marked cells are described in detail.

Animals↗