scholarly journals Messenger RNA transcripts of the meiotic regulator BOULE in the testis of azoospermic men and their application in predicting the success of sperm retrieval

2005 ◽  
Vol 20 (3) ◽  
pp. 782-788 ◽  
Author(s):  
Yung Ming Lin ◽  
Pao Lin Kuo ◽  
Ying Hung Lin ◽  
Yen Ni Teng ◽  
Johnny Shinn Nan Lin
Keyword(s):  
Messenger Rna ◽  
Sperm Retrieval ◽  
Rna Transcripts

The Maize Stripe Virus Major Noncapsid Protein Messenger RNA Transcripts Contain Heterogeneous Leader Sequences at Their 5′ Termini

Virology ◽  
1993 ◽  
Vol 197 (2) ◽  
pp. 808-812 ◽  
Author(s):  
Layne Huiet ◽  
Paul A. Feldstein ◽  
James H. Tsai ◽  
Bryce W. Falk
Keyword(s):  
Messenger Rna ◽  
Rna Transcripts

NEW APPROACHES TOWARDS FLUORESCENCE LABELLING OF MESSENGER RNA TRANSCRIPTS

2001 ◽  
Vol 20 (4-7) ◽  
pp. 1181-1185 ◽  
Author(s):  
Arlette Garnier ◽  
Dieter Hüsken ◽  
Jan Weiler
Keyword(s):  
Messenger Rna ◽  
Fluorescence Labelling ◽  
Rna Transcripts ◽  
New Approaches

scholarly journals Human neutrophils express the alpha 1-antitrypsin gene and produce alpha 1-antitrypsin

Blood ◽  
1991 ◽  
Vol 77 (12) ◽  
pp. 2724-2730
Author(s):  
RM du Bois ◽  
JF Bernaudin ◽  
P Paakko ◽  
R Hubbard ◽  
H Takahashi ◽  
...  
Keyword(s):  
Messenger Rna ◽  
De Novo ◽  
Sodium Dodecyl ◽  
Mononuclear Phagocytes ◽  
Northern Analysis ◽  
Human Neutrophils ◽  
Normal Tissues ◽  
Rna Transcripts ◽  
Neutrophil Degranulation ◽  
Alpha 1 Antitrypsin

The potent serine protease, neutrophil elastase (NE), is stored in neutrophil azurophilic granules, where it is available to degrade phagocytosed material and can be released by the cell to assist in tissue migration and help clear tissue debris. While neutrophils carry NE, they cannot produce it; the NE gene is expressed only in bone marrow granulocyte precursor cells. Protection of normal tissues from the destructive capacity of NE is provided by alpha 1-antitrypsin (alpha 1 AT), a 52-Kd serine antiprotease produced by hepatocytes and mononuclear phagocytes. In the context of the broad destructive capacity of NE, we evaluated the concept that human neutrophils may be able to modulate the extracellular activity of NE by synthesizing and secreting alpha 1AT. Immunocytochemical analysis demonstrated that the neutrophil contains alpha 1AT. Northern analysis and in situ hybridization with alpha 1AT-specific probes demonstrated the presence of alpha 1AT messenger RNA transcripts within neutrophils. [35S]methionine-labeling of neutrophils followed by immunoprecipitation of the supernatant with an anti-alpha 1AT antibody and sodium dodecyl sulfate-acrylamide gel analysis demonstrated that neutrophils can synthesize alpha 1AT de novo and secrete the synthesized molecule. In the presence of major neutrophil degranulation, the antiprotease effect of neutrophil alpha 1AT is overwhelmed, allowing the NE to act unopposed in the extracellular microenvironment. However, in conditions where small amounts of NE are released by neutrophils, at least some of the secreted newly synthesized alpha 1AT was capable of complexing with NE. Thus, despite the fact that the neutrophil cannot synthesize NE, it can synthesize and secrete alpha 1AT, the inhibitor of NE, ie, the neutrophil is capable, to some extent, of modulating NE activity in the local milieu without the help of antiproteases produced by other cells.

ALTERED ABUNDANCE OF MESSENGER RNA TRANSCRIPTS WHOSE PRODUCTS HAVE MEMBRANE ATTACHMENT SITES IN PREGNANCY-INDUCED HYPERTENSION

2002 ◽  
Vol 10 (04) ◽  
pp. 447-461 ◽  
Author(s):  
GREGORY I. C. SIMPSON ◽  
LESLIE C. SHARKEY ◽  
JOHN FRAY
Keyword(s):  
Gene Expression ◽  
Oral Administration ◽  
Messenger Rna ◽  
Gene Products ◽  
Microarray Technology ◽  
Rna Transcripts ◽  
Attachment Sites ◽  
Glycosylation Sites ◽  
Induced Hypertension ◽  
Membrane Attachment

Pregnancy-induced hypertensive disorders (PIH) are leading causes of maternal mortality. Although the mechanism responsible for initiating and maintaining the disorder is unproven, physiologic molecular attachments in kidney and placenta play a role. The SHHF/Mcc-facp (SHHF) rat has features of the disorder, including abnormal placenta gene expression. To gain a molecular understanding of the gene expression profile associated with PIH, kidneys and placentas of SHHF rats at gestation day 20 were compared to WKY controls using microarray technology. We report that SHHF rats have spontaneous PIH, elevated total placenta weights, and reduced total pup weights than WKY controls and that they also have greater total number of mRNA transcripts expressed in placenta. Kidneys of SHHF rats, on the other hand, not only expressed disproportionately more predicted gene products with attachment sites such as RGD motifs, N-glycosylation sites, and N-myristoylation sites they also responded more profoundly to oral administration of L-arginine. We conclude that the increased abundance of transcripts whose products engage in posttranslational attachments using RGD motifs, N-glycosylation sites, and N-myristoylation sites and the reversal of these increases by oral administration of L-arginine suggests that NO may be of importance in PIH at the level of molecular attachments.

scholarly journals A technical review and guide to RNA fluorescence in situ hybridization

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8806
Author(s):  
Alexander P. Young ◽  
Daniel J. Jackson ◽  
Russell C. Wyeth
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Messenger Rna ◽  
Cultured Cells ◽  
Tissue Preparation ◽  
Functional Roles ◽  
Rna Transcripts ◽  
Methods Development ◽  
Background Removal

RNA-fluorescence in situ hybridization (FISH) is a powerful tool to visualize target messenger RNA transcripts in cultured cells, tissue sections or whole-mount preparations. As the technique has been developed over time, an ever-increasing number of divergent protocols have been published. There is now a broad selection of options available to facilitate proper tissue preparation, hybridization, and post-hybridization background removal to achieve optimal results. Here we review the technical aspects of RNA-FISH, examining the most common methods associated with different sample types including cytological preparations and whole-mounts. We discuss the application of commonly used reagents for tissue preparation, hybridization, and post-hybridization washing and provide explanations of the functional roles for each reagent. We also discuss the available probe types and necessary controls to accurately visualize gene expression. Finally, we review the most recent advances in FISH technology that facilitate both highly multiplexed experiments and signal amplification for individual targets. Taken together, this information will guide the methods development process for investigators that seek to perform FISH in organisms that lack documented or optimized protocols.

The organization of transcription on lampbrush chromosomes

1978 ◽  
Vol 283 (997) ◽  
pp. 359-366 ◽  
Keyword(s):  
Messenger Rna ◽  
Repetitive Sequences ◽  
Simple Explanation ◽  
Lampbrush Chromosomes ◽  
Chromosomal Dna ◽  
Rna Sequences ◽  
Nonhistone Proteins ◽  
Rna Transcripts ◽  
Biochemical Studies ◽  
Regular Arrays

The meiotic lampbrush chromosomes of amphibian oocytes display readily distinguishable regions of transcription (lateral loops) which extend from axial condensates of chromatin (chromomeres). The chromomeres contain most of the chromosomal DNA which, along with histone, is tightly compacted as regular arrays of DNP. Many RNA transcripts are generated on the lateral loops, and heterogeneous nonhistone proteins associate with these transcripts, forming periodic condensates of 20-30 nm ribonucleoprotein (RNP) particles. These unit particles aggregate in various ways and to varying degrees and thereby confer distinctive gross morphologies to particular loops. There are about 10 4 lateral loops per haploid complement of newt chromosomes and this figure is similar to the experimentally derived number of different messenger RNA sequences found in oocytes. From cytological and biochemical studies it is now possible to consider individual lateral loops from various aspects: as morphologically distinct units; as units of inheritance; as units of functional activity; as units of transcription; as units of transcribed repetitive sequences; and as units containing one coding sequence. The difficulties in arriving at a simple explanation of the organization of transcription in lampbrush chromosomes are discussed.

Stability of RNA in developing Xenopus embryos and identification of a destabilizing sequence in TFIIIA messenger RNA

Development ◽  
1988 ◽  
Vol 102 (4) ◽  
pp. 837-852 ◽  
Author(s):  
R. Harland ◽  
L. Misher
Keyword(s):  
Xenopus Laevis ◽  
Half Life ◽  
Messenger Rna ◽  
Untranslated Region ◽  
Circular Rna ◽  
Midblastula Transition ◽  
Rna Transcripts ◽  
Xenopus Embryos ◽  
A Site

Synthetic capped RNA transcripts injected into fertilized eggs of Xenopus laevis have a half-life of 3–4 h. Addition of a long (approximately 200 nucleotide) poly(A) tail increases the half-life to 6–8 h which approaches the half-life of natural polyadenylated globin RNA injected into embryos. Since exonucleolytic action alone could account for the degradation of RNA, we tested whether circular RNA is stable after injection and find that circles are exceptionally stable (half-life greater than 40 h). After the midblastula transition, polyadenylated chloramphenicol transferase (CAT) mRNAs transcribed from injected plasmids have a half-life of 2.5 h. Insertion of a 1000 nucleotide 3′ untranslated region from the Xhox-36 gene into the transcripts does not affect the half-life. In contrast to the finding that internal sequences do not affect stability, we find that sequences from the TFIIIA message reduce the half-life of CAT mRNA from 2.5 h to less than 30 min. We conclude that most RNAs are degraded exonucleolytically from the 3′ end, but specialized internal sequences can greatly destabilize the RNA, possibly by acting as a site for an endonuclease.

scholarly journals Human neutrophils express the alpha 1-antitrypsin gene and produce alpha 1-antitrypsin

Blood ◽  
1991 ◽  
Vol 77 (12) ◽  
pp. 2724-2730 ◽  
Author(s):  
RM du Bois ◽  
JF Bernaudin ◽  
P Paakko ◽  
R Hubbard ◽  
H Takahashi ◽  
...  
Keyword(s):  
Messenger Rna ◽  
De Novo ◽  
Sodium Dodecyl ◽  
Mononuclear Phagocytes ◽  
Northern Analysis ◽  
Human Neutrophils ◽  
Normal Tissues ◽  
Rna Transcripts ◽  
Neutrophil Degranulation ◽  
Alpha 1 Antitrypsin

Abstract The potent serine protease, neutrophil elastase (NE), is stored in neutrophil azurophilic granules, where it is available to degrade phagocytosed material and can be released by the cell to assist in tissue migration and help clear tissue debris. While neutrophils carry NE, they cannot produce it; the NE gene is expressed only in bone marrow granulocyte precursor cells. Protection of normal tissues from the destructive capacity of NE is provided by alpha 1-antitrypsin (alpha 1 AT), a 52-Kd serine antiprotease produced by hepatocytes and mononuclear phagocytes. In the context of the broad destructive capacity of NE, we evaluated the concept that human neutrophils may be able to modulate the extracellular activity of NE by synthesizing and secreting alpha 1AT. Immunocytochemical analysis demonstrated that the neutrophil contains alpha 1AT. Northern analysis and in situ hybridization with alpha 1AT-specific probes demonstrated the presence of alpha 1AT messenger RNA transcripts within neutrophils. [35S]methionine-labeling of neutrophils followed by immunoprecipitation of the supernatant with an anti-alpha 1AT antibody and sodium dodecyl sulfate-acrylamide gel analysis demonstrated that neutrophils can synthesize alpha 1AT de novo and secrete the synthesized molecule. In the presence of major neutrophil degranulation, the antiprotease effect of neutrophil alpha 1AT is overwhelmed, allowing the NE to act unopposed in the extracellular microenvironment. However, in conditions where small amounts of NE are released by neutrophils, at least some of the secreted newly synthesized alpha 1AT was capable of complexing with NE. Thus, despite the fact that the neutrophil cannot synthesize NE, it can synthesize and secrete alpha 1AT, the inhibitor of NE, ie, the neutrophil is capable, to some extent, of modulating NE activity in the local milieu without the help of antiproteases produced by other cells.

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