scholarly journals Sorting of the Specific Granule Protein, NGAL, During Granulocytic Maturation of HL-60 Cells

Blood ◽  
1997 ◽  
Vol 89 (6) ◽  
pp. 2113-2121 ◽  
Author(s):  
Véronique Le Cabec ◽  
Jero Calafat ◽  
Niels Borregaard
Keyword(s):  
Human Neutrophil ◽  
Cell Stage ◽  
Cytomegalovirus Promoter ◽  
Granule Protein ◽  
Transport Vesicles ◽  
Different Types ◽  
Storage Granules ◽  
Specific Granule ◽  
Granule Proteins

Abstract The different types of human neutrophil granules (azurophil, specific, and gelatinase granules) are formed sequentially during maturation of neutrophils from the promyelocyte stage to the band cell stage. The promyelocytic HL-60 cells can maturate to segmented granulocytes but are incapable of activating the transcription of any known intragranular protein, normally located in specific or gelatinase granules. To study the sorting of granule proteins during maturation, we transfected HL-60 cells with the specific granule protein NGAL, inserted under control of a cytomegalovirus promoter. We previously showed that NGAL is sorted to azurophil granules and colocalizes with myeloperoxidase in undifferentiated HL-60 cells. We show here that, when such transfected HL-60 cells differentiate into granulocytes, newly synthesized NGAL is not retained in granules but is constitutively secreted. This indicates that highly specific mechanisms must exist that are responsible for diverting transport vesicles into storage granules, and that HL-60 cells not only lack the ability to activate transcription of specific granule proteins, but also lose the ability to form storage granules during maturation.

The analysis of chromatin organisation allows selection of mouse antral oocytes competent for development to blastocyst

Zygote ◽  
2002 ◽  
Vol 10 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Maurizio Zuccotti ◽  
Rubén H. Ponce ◽  
Michele Boiani ◽  
Stefano Guizzardi ◽  
Paolo Govoni ◽  
...  
Keyword(s):  
Developmental Competence ◽  
Farm Animals ◽  
Cell Stage ◽  
Developmental Potential ◽  
Different Types ◽  
Chromatin Organisation ◽  
Selection For ◽  
Gamete Selection ◽  
Selection Of

Mouse antral oocytes can be classified in two different types termed SN or NSN oocytes, depending on the presence or absence, respectively, of a ring of Hoechst 33342-positive chromatin surrounding the nucleolus. The aim of the present study was to test the developmental competence to blastocyst of the two types of oocytes. Here we show that following isolation, classification and culture of cumulus-free antral oocytes, 14.7% and 74.5% of NSN and SN oocytes, respectively, reached the metaphase II stage. When fertilised and further cultured none of the metaphase II NSN oocytes developed beyond the 2-cell stage whilst 47.4% of the metaphase II SN oocytes reached the 4-cell stage and 18.4% developed to blastocyst. The findings reported in this paper may contribute to improved procedures of female gamete selection for in vitro fertilisation of humans and farm animals. Furthermore, the selection of oocytes with better developmental potential may be of interest for studies on nuclear/cytoplasm interaction, particularly in nuclear-transfer experiments.

Intracellular activation of the fibrinolytic cascade in the Quebec Platelet Disorder

2003 ◽  
Vol 90 (08) ◽  
pp. 293-298 ◽  
Author(s):  
Prameet Sheth ◽  
Walter Kahr ◽  
Anwar Haq ◽  
Dragoslava Veljkovic ◽  
Georges Rivard ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Small Proportion ◽  
Bleeding Disorder ◽  
Phenotypic Changes ◽  
Granule Protein ◽  
Type Plasminogen Activator ◽  
Urokinase Type Plasminogen Activator ◽  
Platelet Disorder ◽  
Granule Proteins

SummaryThe Quebec Platelet Disorder (QPD) is an unusual bleeding disorder associated with increased platelet stores of urokinase-type plasminogen activator (u-PA) and proteolysis of platelet α–granule proteins. The increased u-PA and proteolyzed plasmino-gen in QPD platelets led us to investigate possible contributions of intracellular plasmin generation to QPD α-granule proteolysis. ELISA indicated there were normal amounts of plasminogen and plasmin-α2-antiplasmin (PAP) complexes in QPD plasmas. Like normal platelets, QPD platelets contained only a small proportion of the blood plasminogen, however, they contained an increased amount of PAP complexes compared to normal platelets (P < 0.005). The quantities of plasminogen stored in platelets were important to induce QPD-like proteolysis of normal α-granule proteins by two chain u-PA (tcu-PA) in vitro. Moreover, adding supplemental plasminogen to QPD, but not to control, platelet lysates, triggered further α-granule protein proteolysis to forms that comigrated with plasmin degraded proteins. These data suggest the generation of increased but limiting amounts of plasmin within platelets is involved in producing the unique phenotypic changes to α-granule proteins in QPD platelets. The QPD is the only known bleeding disorder associated with chronic, intracellular activation of the fibrinolytic cascade.

Studies of α-granule proteins in cultured human megakaryocytes

2003 ◽  
Vol 90 (11) ◽  
pp. 844-852 ◽  
Author(s):  
Dragoslava Veljkovic ◽  
Elisabeth Cramer ◽  
Gulie Alimardani ◽  
Serge Fichelson ◽  
Jean-Marc Massé ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Protein Processing ◽  
Platelet Factor ◽  
Protein Storage ◽  
Granule Protein ◽  
Granule Proteins ◽  
Processing And Storage ◽  
Von Willebrand ◽  
And Storage ◽  
Willebrand Factor

Summaryα-Granule protein storage is important for producing platelets with normal haemostatic function. The low to undetectable levels of several megakaryocyte-synthesized α-granule proteins in normal plasma suggest megakaryocytes are important to sequester these proteins in vivo. α-Granule protein storage in vitrohas been studied using other cell types, with differences observed in how some proteins are processed compared to platelets. Human megakaryocytes, cultured from cord blood CD34+cells and grown in serum-free media containing thrombopoietin, were investigated to determine if they could be used as a model for studying normal α-granule protein processing and storage. ELISA indicated that cultured megakaryocytes contained the α-granule proteins multimerin, von Willebrand factor, thrombospondin-1, β-thromboglobulin and platelet factor 4, but no detectable fibrinogen and factor V. A significant proportion of the α-granule protein in megakaryocyte cultures was contained within the cells (averages: 41 – 71 %), consistent with storage. Detailed analyses of multimerin and von Willebrand factor confirmed that α-granule proteins were processed to mature forms and were predominantly located in the α-granules of cultured megakaryocytes. Thrombopoietin-stimulated cultured megakaryocytes provide a useful model for studying α-granule protein processing and storage.

scholarly journals Neutrophil-specific Granule Deficiency Results from a Novel Mutation with Loss of Function of the Transcription Factor CCAAT/Enhancer Binding Protein ε

1999 ◽  
Vol 189 (11) ◽  
pp. 1847-1852 ◽  
Author(s):  
Julie A. Lekstrom-Himes ◽  
Susan E. Dorman ◽  
Piroska Kopar ◽  
Steven M. Holland ◽  
John I. Gallin
Keyword(s):  
Novel Mutation ◽  
Binding Protein ◽  
Loss Of Function ◽  
Dimerization Domain ◽  
Knockout Mouse Model ◽  
Enhancer Binding Protein ◽  
Ccaat Enhancer Binding Protein ◽  
Specific Granule ◽  
Granule Proteins ◽  
Secondary Granule

Neutrophil-specific granule deficiency (SGD) is a rare disorder characterized by recurrent pyogenic infections, defective neutrophil chemotaxis and bactericidal activity, and lack of neutrophil secondary granule proteins. CCAAT/enhancer binding protein (C/EBP)ε, a member of the leucine zipper family of transcription factors, is expressed primarily in myeloid cells, and its knockout mouse model possesses distinctive defects, including a lack of neutrophil secondary granule proteins. Sequence analysis of the genomic DNA of a patient with SGD revealed a five-basepair deletion in the second exon of the C/EBPε locus. The predicted frame shift results in a truncation of the 32-kD major C/EBPε isoform, with loss of the dimerization domain, DNA binding region, and transcriptional activity. The multiple functional defects observed in these early neutrophil progenitor cells, a consequence of C/EBPε deficiency, define SGD as a defect in myelopoiesis and establish the requirement for C/EBPε for the promyelocyte–myelocyte transition in myeloid differentiation.

Expression of Azurophil and specific granule proteins during differentiation of NB4 cells in neutrophils

1998 ◽  
Vol 175 (2) ◽  
pp. 203-210 ◽  
Author(s):  
Christine Grégoire ◽  
Heidi Welch ◽  
Catherine Astarie-Dequeker ◽  
Isabelle Maridonneau-Parini
Keyword(s):  
Nb4 Cells ◽  
Specific Granule ◽  
Granule Proteins

Neutrophil-specific granule deficiency: homozygous recessive inheritance of a frameshift mutation in the gene encoding transcription factor CCAAT/enhancer binding protein–ε

Blood ◽  
2001 ◽  
Vol 97 (9) ◽  
pp. 2561-2567 ◽  
Author(s):  
Adrian F. Gombart ◽  
Masaaki Shiohara ◽  
Scott H. Kwok ◽  
Kazunaga Agematsu ◽  
Atsushi Komiyama ◽  
...  
Keyword(s):  
Binding Protein ◽  
Genetic Defect ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Antimicrobial Protein ◽  
Homozygous Mutation ◽  
Recessive Inheritance ◽  
Granule Protein ◽  
Enhancer Binding Protein ◽  
Specific Granule

Abstract Neutrophil-specific granule deficiency (SGD) is a rare congenital disorder. The neutrophils of individuals with SGD display atypical bi-lobed nuclei, lack expression of all secondary and tertiary granule proteins, and possess defects in chemotaxis, disaggregation, receptor up-regulation, and bactericidal activity, resulting in frequent and severe bacterial infections. Previously, a homozygous mutation in theCCAAT/enhancer binding protein–ε (C/EBPε) gene was reported for one case of SGD. To substantiate the role of C/EBPε in the development of SGD and elucidate its mechanism of inheritance, the mutational status of the gene was determined in a second individual. An A-nucleotide insertion in the coding region of the C/EBPε gene was detected. This mutation completely abolished the predicted translation of all C/EBPε isoforms. Microsatellite and nucleotide sequence analyses of the C/EBPε locus in the parents of the proband indicated that the disorder may have resulted from homozygous recessive inheritance of the mutant allele from an ancestor shared by both parents. The mutant C/EBPε32 protein localized in the cytoplasm rather than the nucleus and was unable to activate transcription. Consistent with this, a significant decrease in the levels of the messenger RNAs (mRNAs) encoding the secondary granule protein human 18-kd cationic antimicrobial protein (hCAP-18)/LL-37 and the primary granule protein bactericidal/permeability-increasing protein were observed in the patient. The hCAP-18 mRNA was induced by overexpression of C/EBPε32 in the human myeloid leukemia cell line, U937, supporting the hypothesis that C/EBPε is a key regulator of granule gene synthesis. This study strongly implicates mutation of theC/EBPε gene as the primary genetic defect involved in the development of neutrophil SGD and defines its mechanism of inheritance.

Comparison of the numbers of plasmalemmal vesicles in arterial and venous endothelia of rats

1986 ◽  
Vol 227 (1246) ◽  
pp. 17-20 ◽  
Keyword(s):  
Plasma Proteins ◽  
Femoral Vein ◽  
Vena Cava ◽  
Ruthenium Red ◽  
Electron Microscopic ◽  
Plasmalemmal Vesicles ◽  
Transport Vesicles ◽  
Different Types ◽  
Order Of Magnitude ◽  
Selective Accumulation

The numbers of plasmalemmal vesicles in endothelial cells of rat blood vessels were determined on electron microscopic sections. In all vessels examined which included aorta and carotid and femoral arteries, vena cava and femoral vein, and lung and brain capillaries, the numbers were of the same order of magnitude. For arteries the numbers were about double those for the corresponding veins. About one-third of all vesicles could be stained with ruthenium red after its infusion into the vessels. The results make it improbable that differences in numbers of ‘transport’ vesicles in different types of blood vessel contribute significantly to the selective accumulation of atherogenic plasma proteins in arteries.

Deficiency of the specific granule proteins, R-binder/transcobalamin I and lactoferrin, in plasma and saliva: A new disorder

2001 ◽  
Vol 100 (2) ◽  
pp. 145-151 ◽  
Author(s):  
Judith C. Lin ◽  
Niels Borregaard ◽  
Howard A. Liebman ◽  
Ralph Carmel
Keyword(s):  
Specific Granule ◽  
Granule Proteins ◽  
Transcobalamin I
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