scholarly journals Different roles for two enhancer domains in the organ- and age-specific pattern of polyomavirus replication in the mouse.

1992 ◽  
Vol 12 (8) ◽  
pp. 3628-3635 ◽  
Author(s):  
A Amalfitano ◽  
L G Martin ◽  
M M Fluck
Keyword(s):  
Transcription Factors ◽  
In Situ Hybridization ◽  
Mammary Gland ◽  
Crucial Role ◽  
Specific Pattern ◽  
Neonatal Stage ◽  
Murine Polyomavirus ◽  
A Domains

Viral replication in mice infected with murine polyomavirus strains with novel enhancer rearrangements was analyzed by direct in situ hybridization of whole mouse sections and by hybridization of nucleic acids extracted from a specific set of organs. The enhancer rearrangements included a deletion of the B domain as well as duplications within the A domain. Comparisons between enhancer variants demonstrate that the B domain plays an important role in replication in most organs, in particular in the kidney, at the neonatal stage (days 0 to 7 postbirth). In contrast, the B domain is not required in those organs which can sustain replication in the adult, i.e. mammary gland, skin, and bone (class I organs [J. J. Wirth, A. Amalfitano, R. Gross, M. B. A. Oldstone, and M. M. Fluck, J. Virol. 66:3278-3286, 1992]). Altogether, the results suggest that the B and A domains mediate very different functions in infection of mice, controlling the acute and persistent phases of infection, respectively. A model of mouse infection based on the crucial role of differentially expressed host transcription factors is presented.

Different roles for two enhancer domains in the organ- and age-specific pattern of polyomavirus replication in the mouse

1992 ◽  
Vol 12 (8) ◽  
pp. 3628-3635
Author(s):  
A Amalfitano ◽  
L G Martin ◽  
M M Fluck
Keyword(s):  
Transcription Factors ◽  
In Situ Hybridization ◽  
Mammary Gland ◽  
Crucial Role ◽  
Specific Pattern ◽  
Neonatal Stage ◽  
Murine Polyomavirus ◽  
A Domains

Viral replication in mice infected with murine polyomavirus strains with novel enhancer rearrangements was analyzed by direct in situ hybridization of whole mouse sections and by hybridization of nucleic acids extracted from a specific set of organs. The enhancer rearrangements included a deletion of the B domain as well as duplications within the A domain. Comparisons between enhancer variants demonstrate that the B domain plays an important role in replication in most organs, in particular in the kidney, at the neonatal stage (days 0 to 7 postbirth). In contrast, the B domain is not required in those organs which can sustain replication in the adult, i.e. mammary gland, skin, and bone (class I organs [J. J. Wirth, A. Amalfitano, R. Gross, M. B. A. Oldstone, and M. M. Fluck, J. Virol. 66:3278-3286, 1992]). Altogether, the results suggest that the B and A domains mediate very different functions in infection of mice, controlling the acute and persistent phases of infection, respectively. A model of mouse infection based on the crucial role of differentially expressed host transcription factors is presented.

scholarly journals On the Origin of Tetraploid Vernal Grasses (Anthoxanthum) in Europe

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 966
Author(s):  
Zuzana Chumová ◽  
Terezie Mandáková ◽  
Pavel Trávníček
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Crucial Role ◽  
Evolutionary History ◽  
Essential Role ◽  
Grass Species ◽  
Polyploid Species ◽  
Anthoxanthum Odoratum ◽  
European Populations

Polyploidy has played a crucial role in the evolution of many plant taxa, namely in higher latitudinal zones. Surprisingly, after several decades of an intensive research on polyploids, there are still common polyploid species whose evolutionary history is virtually unknown. Here, we addressed the origin of sweet vernal grass (Anthoxanthum odoratum) using flow cytometry, DNA sequencing, and in situ hybridization-based cytogenetic techniques. An allotetraploid and polytopic origin of the species has been verified. The chromosome study reveals an extensive variation between the European populations. In contrast, an autopolyploid origin of the rarer tetraploid vernal grass species, A. alpinum, has been corroborated. Diploid A. alpinum played an essential role in the polyploidization of both European tetraploids studied.

scholarly journals Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

1994 ◽  
Vol 42 (9) ◽  
pp. 1271-1276 ◽  
Author(s):  
M Numata ◽  
T Ono ◽  
S Iseki
Keyword(s):  
In Situ Hybridization ◽  
Significant Role ◽  
Meiotic Prophase ◽  
Oligonucleotide Probe ◽  
Mouse Testis ◽  
Seminiferous Tubules ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

Sequential silver staining and in situ hybridization reveal a direct association between rDNA levels and the expression of homologous nucleolar organizing regions: a hypothesis for NOR structure and function

1998 ◽  
Vol 111 (10) ◽  
pp. 1433-1439
Author(s):  
F. Zurita ◽  
R. Jimenez ◽  
M. Burgos ◽  
R.D. de la Guardia
Keyword(s):  
Transcription Factors ◽  
In Situ Hybridization ◽  
Silver Staining ◽  
Organizer Region ◽  
Nucleolar Organizer ◽  
Interindividual Variation ◽  
Nucleolar Organizer Regions ◽  
Single Pair ◽  
Ribosomal Cistrons

We have developed a procedure for sequential silver staining and in situ hybridization to analyze the relationship between the amount of rDNA present in nucleolar organizer regions, as estimated by in situ hybridization, and their level of expression, as estimated by the silver signal. For simplicity we used cells from the insectivorous mole Talpa occidentalis, which have a single pair of nucleolar organizer regions in chromosome pair 3. The relative content of ribosomal cistrons was also related to the hierarchy of activation of the nucleolar organizer regions present in this chromosomal pair. Statistical analyses demonstrated that both the relative level of expression and the activation hierarchy depended mainly on the number of ribosomal cistrons in nucleolar organizer regions. We propose a functional two-step hypothesis, which is consistent with most known data concerning interchromosomal, intercellular and interindividual variation in a number of plant and animal species, including Talpa occidentalis. In step one, the first available transcription factors bind randomly to the ribosomal promoters, such that larger nucleolar organizer regions are more likely to recruit them. In the second step the remaining transcription factors are recruited in a cooperative way, thus completing activation of one nucleolar organizer region, before the next one becomes active.

scholarly journals Expression and role of serum and glucocorticoid-regulated kinase 2 in the regulation of Na+/H+ exchanger 3 in the mammalian kidney

2010 ◽  
Vol 299 (6) ◽  
pp. F1496-F1506 ◽  
Author(s):  
Alan C. Pao ◽  
Aditi Bhargava ◽  
Francesca Di Sole ◽  
Raymond Quigley ◽  
Xinli Shao ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Proximal Tubule ◽  
Cell Surface Expression ◽  
Thick Ascending Limb ◽  
Proximal Tubule Cells ◽  
Mammalian Kidney ◽  
Tubule Cells ◽  
Exchange Activity

Serum and glucocorticoid-regulated kinase 2 (sgk2) is 80% identical to the kinase domain of sgk1, an important mediator of mineralocorticoid-regulated sodium (Na+) transport in the distal nephron of the kidney. The expression pattern and role in renal function of sgk2 are virtually uncharacterized. In situ hybridization and immunohistochemistry of rodent kidney coupled with real-time RT-PCR of microdissected rat kidney tubules showed robust sgk2 expression in the proximal straight tubule and thick ascending limb of the loop of Henle. Sgk2 expression was minimal in distal tubule cells with aquaporin-2 immunostaining but significant in proximal tubule cells with Na+/H+ exchanger 3 (NHE3) immunostaining. To ascertain whether mineralocorticoids regulate expression of sgk2 in a manner similar to sgk1, we examined sgk2 mRNA expression in the kidneys of adrenalectomized rats treated with physiological doses of aldosterone together with the glucocorticoid receptor antagonist RU486. Northern blot analysis and in situ hybridization showed that, unlike sgk1, sgk2 expression in the kidney was not altered by aldosterone treatment. Based on the observation that sgk2 is expressed in proximal tubule cells that also express NHE3, we asked whether sgk2 regulates NHE3 activity. We heterologously expressed sgk2 in opossum kidney (OKP) cells and measured Na+/H+ exchange activity by Na+-dependent cell pH recovery. Constitutively active sgk2, but not sgk1, stimulated Na+/H+ exchange activity by >30%. Moreover, the sgk2-mediated increase in Na+/H+ exchange activity correlated with an increase in cell surface expression of NHE3. Together, these results suggest that the pattern of expression, regulation, and role of sgk2 within the mammalian kidney are distinct from sgk1 and that sgk2 may play a previously unrecognized role in the control of transtubular Na+ transport through NHE3 in the proximal tubule.

scholarly journals In Situ Hybridization Analysis of ZPK Gene Expression During Murine Embryogenesis

1997 ◽  
Vol 45 (1) ◽  
pp. 107-118 ◽  
Author(s):  
André Nadeau ◽  
Gilles Grondin ◽  
Richard Blouin
Keyword(s):  
In Situ Hybridization ◽  
Northern Blot Analysis ◽  
Spatial And Temporal Patterns ◽  
Serine Threonine Kinase ◽  
Teratocarcinoma Cell ◽  
Threonine Kinase ◽  
Cell Lineages ◽  
Polymerase Chain

ZPK is a recently described protein serine/threonine kinase that has been originally identified from a human teratocarcinoma cell line by the polymerase chain reaction and whose function in signal transduction has not yet been elucidated. To investigate the potential role of this protein kinase in developmental processes, we have analyzed the spatial and temporal patterns of expression of the ZPK gene in mouse embryos of different gestational ages. Northern blot analysis revealed a single mRNA species of about 3.5 KB from Day 11 of gestation onwards. In situ hybridization studies demonstrated strong expression of ZPK mRNA in brain and in a variety of embryonic organs that rely on epithelio-mesenchymal interactions for their development, including skin, intestine, pancreas, and kidney. In these tissues, the ZPK mRNA was localized primarily in areas composed of specific types of differentiating cells, and this expression appeared to be upregulated at a time concomitant with the onset of terminal differentiation. Taken together, these observations raise the possibility that the ZPK gene product is involved in the establishment and/or maintenance of a fully cytodifferentiated state in a variety of cell lineages.

scholarly journals Transcriptome-Wide Analyses Provide Insights into Development of the Hedychium Coronarium Flower, Revealing Potential Roles of PTL

2020 ◽  
Author(s):  
Tong Zhao ◽  
Alma Piñeyro-Nelson ◽  
Qianxia Yu ◽  
Xiaoying Hu ◽  
Huanfang Liu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Flower Development ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Floral Organ ◽  
Mrna In Situ Hybridization ◽  
Hedychium Coronarium ◽  
Organ Identity

Abstract Background:The flower of Hedychium coronarium possesses highly specialized floral organs: a synsepalous calyx, petaloid staminodes and a labellum. The formation of these organs is controlled by two gene categories: floral organ identity genes and organ boundary genes, which may function individually or jointly during flower development. Although the floral organogenesis of H. coronarium has been studied at the morphological level, the underlying molecular mechanisms involved in its floral development still remain poorly understood. In addition, previous works analyzing the role of MADS-box genes in controlling floral organ specification in some Zingiberaceae did not address the molecular mechanisms involved in the formation of particular organ morphologies that emerge later in flower development, such as the synsepalous calyx formed through intercalary growth of adjacent sepals. Results:Here, we used comparative transcriptomics combined with Real-time quantitative PCR and mRNA in situ hybridization to investigate gene expression patterns of ABC-class genes in H. coronarium flowers, as well as the homolog of the organ boundary gene PETAL LOSS (HcPTL). qRT-PCR detection showed that HcAP3 and HcAG were expressed in both the petaloid staminode and the fertile stamen. mRNA in situ hybridization showed that HcPTL was expressed in developing meristems, including cincinnus primordia, floral primordia, common primordia and almost all new initiating floral organ primordia.Conclusions:Our studies found that stamen/petal identity or stamen fertility in H. coronarium was not necessarily correlated with the differential expression of HcAP3 and HcAG. We also found a novel spatio-temporal expression pattern for HcPTL mRNA, suggesting it may have evolved a lineage-specific role in the morphogenesis of the Hedychium flower. Our study provides a new transcriptome reference and a functional hypothesis regarding the role of a boundary gene in organ fusion that should be further addressed through phylogenetic analyzes of this gene, as well as functional studies.

Gonadal expression of c-kit encoded at the W locus of the mouse

Development ◽  
1990 ◽  
Vol 110 (4) ◽  
pp. 1057-1069 ◽  
Author(s):  
K. Manova ◽  
K. Nocka ◽  
P. Besmer ◽  
R.F. Bachvarova
Keyword(s):  
In Situ Hybridization ◽  
Germ Cells ◽  
Leydig Cells ◽  
Interstitial Tissue ◽  
Type B ◽  
Age Dependence ◽  
Oocyte Growth ◽  
Adult Mice

Recently, it has been shown that the c-kit proto-oncogene is encoded at the white spotting (W) locus in mice. Mutations of this gene cause depletion of germ cells, some hematopoietic cells and melanocytes. In order to define further the role of c-kit in gametogenesis, we have examined its expression in late fetal and postnatal ovaries and in postnatal testis. By RNA blot analysis, c-kit transcripts were not detected in late fetal ovaries but appeared at birth. The relative amount reached a maximum in ovaries of juvenile mice, and decreased in adult ovaries. c-kit transcripts were present in increasing amounts in isolated primordial, growing and full-grown oocytes, as well as in ovulated eggs. Little was detected in early 2-cell embryos and none in blastocysts. In situ hybridization revealed c-kit transcripts in a few oocytes of late fetal ovaries and in all oocytes (from primordial to full-grown) in ovaries from juvenile and adult mice. Expression was also observed in ovarian interstitial tissue from 14 days of age onward. Using indirect immunofluorescence, the c-kit protein was detected on the surface of primordial, growing and full-grown oocytes, as well as on embryos at the 1- and 2-cell stages; little remained in blastocysts. In situ hybridization analysis of testes from mice of different ages demonstrated expression in spermatogonia from 6 days of age onward. Using information provided by determining the stage of the cycle of the seminiferous epithelium for a given tubule and by following the age dependence of labeling, it was concluded that the period of expression of c-kit extends from at least as early as type A2 spermatogonia through type B spermatogonia and into preleptotene spermatocytes. Leydig cells were labelled at all ages examined. The expression pattern in oocytes correlates most strongly with oocyte growth and in male germ cells with gonial proliferation.

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