scholarly journals A Morphometric Analysis of Platelet Dense Granules of Patients with Unexplained Bleeding: A New Entity of Delta-Microgranular Storage Pool Deficiency

2020 ◽  
Vol 9 (6) ◽  
pp. 1734 ◽  
Author(s):  
William T. Gunning ◽  
Meera Raghavan ◽  
Edward P. Calomeni ◽  
James N. Turner ◽  
Bodri Roysam ◽  
...  
Keyword(s):  
Dense Granule ◽  
Control Data ◽  
Dense Granules ◽  
Storage Pool ◽  
Normal Number ◽  
Normal Mean ◽  
The Mean ◽  
Bleeding History ◽  
And Storage

One thousand and eighty patients, having prolonged bleeding times, frequent epistaxis, menorrhagia or easy bruising or other bleeding manifestations, and excluding those with von Willebrand’s disease, were evaluated for platelet dense granule deficiency. The mean diameter of platelet dense granules was determined for all patients using image analysis. Four hundred and ninety-nine had “classic” dense (delta) granule storage pool deficiency (δ-SPD). Five hundred and eighty-one individuals (53.8%) were found to have a normal mean number of dense granules, but for some of these patients, the dense granules were smaller than for the controls. Of the patients having a normal number of dense granules, 165 (28.4%) were found to have significantly smaller granules than the platelets obtained from the control subjects. Their average granule diameter was 123.35 ± 0.86 nm, that is more than three standard deviations below the mean of the control data. Total δ-granule storage pool volumes (TDGV)/platelet were calculated using these measurements. Individuals with δ-SPD had half the number of granules (2.25 ± 0.04 DG/PL) and storage pool volume (3.88 ± 1.06 × 106 nm3) when compared to our control data (4.64 ± 0.11 DG/PL; 10.79 × 106 nm3 ± 0.42). Individuals having a bleeding history but a normal average of small dense granules had a calculated storage pool volume statistically different than controls and essentially the same storage pool volume as patients with δ-SPD. We have identified a sub-classification of δ-SPD that we have defined as micro-granular storage pool deficiency (δ-MGSPD).

Analysis of Platelet Dense Granules in Storage Pool Deficient Patients

1997 ◽  
Vol 3 (S2) ◽  
pp. 73-74
Author(s):  
M.M. McKenzie ◽  
W.T. Gunning ◽  
M.R. Smith ◽  
N.A. Lachant ◽  
J.N. Turner
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Dense Granule ◽  
Dense Granules ◽  
Storage Pool ◽  
Prolonged Bleeding ◽  
Normal Number ◽  
Transmission Electron ◽  
Insight Into ◽  
Whole Mounts

Prolonged bleeding times may be evident in patients with various conditions. Analysis of platelet function can provide an insight into the cause. Platelets contain substances such as ADP, calcium, epinephrine, and serotonin which are stored in the electron dense or delta granules. Our experience has shown normal platelets to have 4-6 dense granules (Figure 1), which is supported by the literature. The prolonged bleeding times of patients who have less than the normal number of dense granules may be attributed to the lack of adequate factors that aid in the clotting process. These storage pool deficient (SPD) patients are being classified by their low dense granule count. Patients with prolonged bleeding times but normal dense granule counts may need to have their granules evaluated for size and volume (Figure 2).Whole mounted platelets were placed in the transmission electron microscope (TEM), and negatives were obtained at a magnification of 10,200x. The typical dense granule seen in normal whole mounts is circular.

scholarly journals Immature dense granules in platelets from mice with platelet storage pool disease

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1300-1306 ◽  
Author(s):  
M Reddington ◽  
EK Novak ◽  
E Hurley ◽  
C Medda ◽  
MP McGarry ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Dense Granule ◽  
Mutant Mice ◽  
Normal Numbers ◽  
Group Of Seven ◽  
Dense Granules ◽  
Storage Pool ◽  
Platelet Storage ◽  
Transmission Electron ◽  
Storage Pool Disease

Mepacrine uptake into platelets and bone marrow megakaryocytes was analyzed to further characterize the dense granule defects in a group of seven mouse pigment mutants that have characteristics of platelet storage pool disease (SPD). In contrast to our previous studies using electron microscopy, this method revealed that all mutants had normal numbers of dense granules. However, total mepacrine uptake in all mutant platelets was significantly diminished to less than 50% of normal uptake. Also, the flashing phenomenon observed when normal dense granules are irradiated with ultraviolet light was either greatly diminished or absent when platelets of individual mutants were similarly irradiated. Therefore the principal defect in the mutant platelets is an inability to accumulate dense granule contents rather than an absence of the granules. Mepacrine uptake into megakaryocytes was indistinguishable in normal and mutant mice. This indicates the mutant dense granule defects appear either very late in megakaryocyte development or early in platelet formation in correlation with development of the mature dense granule. By standard transmission electron microscopy we have not been able to detect gross structural or subcellular abnormalities in either platelets or megakaryocytes of mutant mice. It appears all seven mutants produce immature or functionally abnormal dense granules.

A Functional Basis For The Platelet Storage Pool Deficiency In Chediak-Higashi Cattle

1981 ◽  
Author(s):  
K M Meyers ◽  
M Fukami ◽  
H Holmsen
Keyword(s):  
Freeze Fracture ◽  
Subcellular Fractionation ◽  
Dense Granule ◽  
Bivalent Cation ◽  
Fibrous Matrix ◽  
Dense Granules ◽  
Storage Pool ◽  
Common Basis ◽  
Platelet Storage ◽  
Amine Storage

Platelets from cattle with the morphologic homolog of the Chediak-Higashi (CH) syndrome are essentially devoid of secretable nucleotides and serotonin. There are reduced but still substantial amounts of secretable calcium and magnesium. The storage pool deficiency may be, in part, due to a functional granule defect. Platelets from CH cattle take up serotonin and protect it from degradation for several hours. If these platelets are treated with thrombin, serotonin and bivalent cations are released by mechanisms similar to that of secretion, suggesting a granule location for the released serotonin and cations. This suggestion is verified by subcellular fractionation studies where platelets are first incubated with 14C-serotonin then lysed using a French press. Organelles were then separated on a sucrose gradient by centrifugation. Serotonin in normal bovine platelets is associated with the dense granule or pellet while in CH platelets serotonin is primarily found in a region of the sucrose density zone that in normal platelets contain alpha granules. These findings suggested that some granules in CH platelets are able to acquire the bivalent cation and amine but not the nucleotide component of the bivalent cation-nucleotide-amine storage complex that is thought to occur in normal dense granules.Ultrastructural identification of the serotonin-containing CH granule is not known. There are 2 identifiable granule populations in CH platelets, alpha granules and fibrous matrix granules. Based on serial sectioning freeze fracture and morphometric studies, there are less than 4 of these granules/platelet. Mepacrine studies also demonstrate 2 granule populations. One population has an incidence of 2 per granule and characteristics of normal dense granules. Since the number of fibrous matrix granules and mepacrine granules is similar, a common basis for these granules which has at least some dense granule characteristics, i.e., mepacrine storage, is suggested.

Depletion of Dense Granule Nucleotides during Storage of Human Platelets

1990 ◽  
Vol 63 (02) ◽  
pp. 275-278 ◽  
Author(s):  
D de Korte ◽  
C W N Gouwerok ◽  
R Fijnheer ◽  
R N I Pietersz ◽  
D Roos
Keyword(s):  
Specific Activity ◽  
Adenine Nucleotide ◽  
Human Platelets ◽  
Dense Granule ◽  
Serotonin Content ◽  
Storage Pool ◽  
Diadenosine Triphosphate ◽  
Metabolic Pool ◽  
And Storage ◽  
Hydrolysis Of

SummaryThe energy metabolism of human platelets was studied during storage of platelet concentrates. The platelets were prepared from buffy coats in PVC/DEHP bags and stored for 7 days at room temperature at a concentration of 1.0 × 109/ml with horizontal agitation. The total amount of ATP and ADP decreased with 40% during this storage. This decrease correlated with the disc-tosphere transformation associated with the loss of platelet viability. During storage, the ability to incorporate 3H-adenosine into metabolic ATP and ADP (45 min at 37° C) decreased with 50%. Via measurement of the specific activity of actin-bound ADP and the amount of incorporated radioactivity into total ATP and ADP, we calculated the content of the metabolic and storage pools of ATP and ADP. The results indicate that the decrease in adenine nucleotide levels during storage was mainly caused by a depletion of ATP and ADP from the storage pool, whereas the metabolic pool remained nearly intact. After 7 days, the ATP : ADP ratio of the storage pool had decreased from 1.0 to 0.3, indicating hydrolysis of ATP.Diadenosine-triphosphate and diadenosine-tetraphosphate (present in the storage pool) decreased with only 30%, and the serotonin content remained nearly constant. Therefore, it is unlikely that the storage pool was completely secreted. Probably, the storage pool of nucleotides serves as an internal supply for maintaining the contents of the metabolic pool of ATP and ADP during storage of platelets.

scholarly journals The response of platelets to epinephrine in storage pool deficiency-- evidence pertaining to the role of adenosine diphosphate in mediating primary and secondary aggregation

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1717-1725 ◽  
Author(s):  
HJ Weiss ◽  
B Lages
Keyword(s):  
Platelet Aggregation ◽  
Receptor Agonist ◽  
Adenosine Diphosphate ◽  
Normal Subjects ◽  
Dense Granule ◽  
Binding Studies ◽  
Dense Granules ◽  
Storage Pool ◽  
Receptor Sites

Abstract Aggregation responses and thromboxane (Tx) formation in ten patients with storage pool deficiency (SPD) specific to the dense granules (delta-SPD) were studied to assess further the role of dense granule adenosine diphosphate (ADP) in mediating platelet aggregation by epinephrine. The ability of epinephrine to elicit secondary aggregation (SA) responses was highly variable in delta-SPD when tested at 5 mumol/L epinephrine, but was consistently abnormal when tested over a range of concentrations. The occurrence of SA in both delta-SPD patients and normal subjects was correlated with the magnitude of the rate of primary aggregation (PA). This PA rate was normal, on average, for the entire patient group but was greater in patients with more consistent SA responses. The PA findings were related to the Kd value obtained in binding studies with 3H-yohimbine, but not with the number of alpha 2-receptor sites. Studies on Tx production (assessed by radioimmunoassay of TxB2) showed that the ability to synthesize Tx from arachidonate was not impaired in delta-SPD, and that there was an absolute positive correlation between epinephrine-induced SA and Tx production. Aggregation in delta-SPD platelets in response to the Tx receptor agonist U44069 was consistently decreased, but could be corrected by addition of ADP. The results of the study suggest that dense granule-derived ADP is not required for PA by epinephrine, but mediates SA as a synergistic agonist with TxA2. This role of ADP in SA may be elucidated more precisely by further studies on platelet activation processes in delta-SPD.

Is the Platelet Involved in the Etiology of Neurocardiogenic Syncope?.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3220-3220
Author(s):  
Cole A. Bennett ◽  
Kathryn E. Boehm ◽  
William T. Gunning
Keyword(s):  
Selective Serotonin Reuptake Inhibitors ◽  
Hospital Admissions ◽  
Adenine Nucleotide ◽  
Vasovagal Syncope ◽  
Dense Granule ◽  
Neurocardiogenic Syncope ◽  
Dense Granules ◽  
Storage Pool ◽  
Number Of Patients ◽  
The Brain

Abstract The purpose of this study was to evaluate platelet dysfunction as a mitigating factor of neurocardiogenic syncope. Neurocardiogenic, or vasovagal syncope as it is also known, is a condition that may lead to temporary loss of consciousness and posture. Commonly referred to as, “fainting,” or, “passing out”, it is the basis for 3% of emergency room visits and 6% of hospital admissions; it is thought to be caused primarily by a lack of blood flow to the brain. However, the pathogenesis of syncope is yet to be completely understood. Vasovagal syncope has been postulated to occur due to a sudden failure of the autonomic nervous system’s ability to maintain blood pressure and heart rate leading to insufficient cerebral perfusion. Subsequently, a cardiac “hyper-contractile” state is created, due to excessive venous pooling and diminished peripheral venous return that is thought to activate afferent mechanoreceptors, interpreted by the brain as hypertension causing a paradoxical reflex of bradycardia, resulting in a drop of peripheral vascular resistance. It has been postulated that an increase in serotonin (5HT) activity in the central nervous system acts as an inhibitor of sympathetic stimulation, preventing the symptoms of syncope. One of a number of pharmacologic approaches to the management of neurocardiogenic syncope is the use of selective serotonin reuptake inhibitors (SSRIs). Neurocardiogenic syncope has also been reported to be a low 5HT disorder. As the major storage pool of 5HT in the circulation is known to exist in platelet (PL)dense granules (DGs), it is possible that a PL dense granule storage pool deficiency (DG SPD) could be an important mediator of neurocardiogenic syncope. We commonly evaluate patients having undiagnosed bleeding symptoms for PL DG SPD. A total of 358 subjects having neurocardiogenic syncope have been evaluated because of a significant bleeding history. Eighty-two percent (293/358) of these patients have DG SPD, with an average of 2.27 ± 0.06 DG/PL (normal: 4–6 DG/PL). Interestingly, 98% (351/358) of these patients are female. Common bleeding symptoms included 53.8% with menorrhagia (189/351), 43.9% (157/358) with easy bruising, and 15.6% with epistaxsis (56/358). It has been reported in a study of 15 patients, that the PL 5HT concentration, determined by HPLC methodology, did not alter during episodes of syncope, suggesting that the serotonergic system does not play a role in the pathophysiology of vasovagal syncope. We are currently evaluating the 5HT concentration extracted from PLs of syncope patients by ELISA. To date, 10 such patients have been studied, having an average of 2.32 ± 0.26 DG/PL; their mean 5HT concentration is 202.13 ± 29.12 ng/109 PL (normal range = 215–850 ng/109 PL). Our preliminary results suggest that a significant number of patients having neurocardiogenic syncope have DG SPD and low concentrations of 5HT stored in these granules. These results may be attributed to the fact that all subjects included in our study presented with a history or symptom of bleeding. The mechanism of 5HT uptake by the platelet is independent of adenine nucleotide and calcium storage within DGs, thus the low 5HT levels found in the few cases described here may/may not be related to pathophysiology of neurocardiogenic syncope. A prospective study of neurocardiogenic syncope patients with/without bleeding histories or symptoms is planned to further evaluate the PL DG storage pool.

Inherited Platelet δ-Storage Pool Disease in Dogs Causing Severe Bleeding: An Animal Model for a Specific ADP Deficiency

1995 ◽  
Vol 74 (03) ◽  
pp. 949-953 ◽  
Author(s):  
Mary Beth Callan ◽  
Joel S Bennett ◽  
Deborah K Phillips ◽  
Mark E Haskins ◽  
James E Hayden ◽  
...  
Keyword(s):  
Adenine Nucleotide ◽  
Bleeding Disorder ◽  
Serotonin Uptake ◽  
Severe Bleeding ◽  
Minor Trauma ◽  
Dense Granules ◽  
Storage Pool ◽  
Normal Number ◽  
Platelet Disorder ◽  
Storage Pool Disease

SummaryThe nature of a disorder producing moderate to severe bleeding after minor trauma, venipuncture, and surgery was studied in 3 families of American cocker spaniel dogs. In the 5 affected dogs tested, platelet counts and measurements of plasma coagulant function and von Willcbrand factor were normal. However, bleeding times were prolonged in 4 of the 5 affected dogs tested, and platelet aggregation in response to ADP and collagen was consistently abnormal in 3, suggesting that the bleeding disorder was due to abnormal platelet function. Measurements of 14C-serotonin uptake and retention by the affected platelets were normal. However, their ADP content was decreased, while their ATP content was normal, resulting in a mean ATP/ADP ratio of 8.32, compared to a mean ratio of 1.9 in normal canine platelets. Electron microscopy revealed that the number and appearance of the dense granules in the affected platelets were indistinguishable from those of normal controls. These studies suggest that this bleeding disorder results from a deficient δ-granule storage pool of ADP; given the normal serotonin uptake and retention by affected platelets and the apparently normal number of dense granules, the ADP deficiency may be the consequence of a selective defect in δ-granule ADP transport. Additional studies of this unique platelet disorder will provide an opportunity to understand the mechanism of adenine nucleotide storage in platelet δ granules.

scholarly journals Immature dense granules in platelets from mice with platelet storage pool disease

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1300-1306 ◽  
Author(s):  
M Reddington ◽  
EK Novak ◽  
E Hurley ◽  
C Medda ◽  
MP McGarry ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Dense Granule ◽  
Mutant Mice ◽  
Normal Numbers ◽  
Group Of Seven ◽  
Dense Granules ◽  
Storage Pool ◽  
Platelet Storage ◽  
Transmission Electron ◽  
Storage Pool Disease

Abstract Mepacrine uptake into platelets and bone marrow megakaryocytes was analyzed to further characterize the dense granule defects in a group of seven mouse pigment mutants that have characteristics of platelet storage pool disease (SPD). In contrast to our previous studies using electron microscopy, this method revealed that all mutants had normal numbers of dense granules. However, total mepacrine uptake in all mutant platelets was significantly diminished to less than 50% of normal uptake. Also, the flashing phenomenon observed when normal dense granules are irradiated with ultraviolet light was either greatly diminished or absent when platelets of individual mutants were similarly irradiated. Therefore the principal defect in the mutant platelets is an inability to accumulate dense granule contents rather than an absence of the granules. Mepacrine uptake into megakaryocytes was indistinguishable in normal and mutant mice. This indicates the mutant dense granule defects appear either very late in megakaryocyte development or early in platelet formation in correlation with development of the mature dense granule. By standard transmission electron microscopy we have not been able to detect gross structural or subcellular abnormalities in either platelets or megakaryocytes of mutant mice. It appears all seven mutants produce immature or functionally abnormal dense granules.

scholarly journals Storage pool deficiency in cattle with the Chediak-Higashi syndrome results from an absence of dense granule precursors in their megakaryocytes

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1726-1734 ◽  
Author(s):  
M Menard ◽  
KM Meyers
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Transmission Electron Microscopy ◽  
Membrane System ◽  
Dense Granule ◽  
Virtual Absence ◽  
Dense Granules ◽  
Storage Pool ◽  
Transmission Electron ◽  
Chediak Higashi Syndrome

Abstract Platelets from cattle with the Chediak-Higashi syndrome (CHS) have a storage pool deficiency and virtual absence of platelet dense granules. Megakaryocytes (MKs) from five control (n = 135) and five CHS (n = 133) cattle were evaluated using standard transmission electron microscopy. Osmiophilic dense granules were not observed in control or CHS MKs. In MKs from normal cattle, clear vesicles of 200- to 650-nm diameter bounded by a sharp membrane were observed. They were easily differentiated from the demarcation membrane system, endoplasmic reticulum, and alpha granules. The clear vesicles were virtually absent in MKs from CHS cattle at all stages of maturation. MKs in bone marrow samples from two control (n = 91) and two CHS (n = 61) cattle that had been processed for the uranaffin reaction were also evaluated. The clear vesicles were replaced by uranaffin-positive granules in MKs from control cattle, but positive uranaffin granules were not observed in CHS MKs. These findings indicate that the platelet dense granule storage pool deficiency in CHS cattle results from an anatomic absence of dense granule precursors in maturing and mature CHS MKs.

scholarly journals The mouse pale ear pigment mutant as a possible animal model for human platelet storage pool deficiency

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 38-43 ◽  
Author(s):  
EK Novak ◽  
SW Hui ◽  
RT Swank
Keyword(s):  
Animal Model ◽  
Human Platelet ◽  
Dense Granule ◽  
Subcellular Organelles ◽  
Dense Granules ◽  
Storage Pool ◽  
Platelet Protein ◽  
Platelet Storage ◽  
Storage Pool Disease ◽  
Experimental Injury

Abstract The mouse pigment mutant pale ear, ep/ep, which has a defect in kidney lysosomal enzyme secretion, had prolonged bleeding on experimental injury. Platelet counts and platelet protein did not differ from normal. There was, however, a deficiency in the platelet dense granule contents, serotonin, ATP, and ADP. Furthermore, a marked reduction of platelet dense granules was observed by electron microscopy. The results suggest that pale ear is a useful animal model in the study of platelet storage pool disease. Studies on this mutant and other pigment mutants have established that one gene can regulate at least three subcellular organelles, including the melanosome, the lysosome, and the platelet dense granule.

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