Characteristics of RNA degradation in the erythroid cell

Burka, Edward R.

doi:10.1172/jci106092

Cited by 36 publications

(40 citation statements)

References 24 publications

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“…5). The obtained sigmoidal relation between signal and age slightly differs from previous studies (6,(8)(9)(10), where RNA was found to be degraded linearly or exponentially. This difference can be due to many factors.…”

Section: Discussioncontrasting

confidence: 98%

“…Presented in the literature, methods for studying RET physiology often involve a difficult to perform isolation and an in vitro measurement of maturation of a synchronized population using radioactive tracers (5,6,8,25). The in vivo determination of the lifespan of RET in blood was determined by studying radioactive iron turnover (5,25).…”

Section: Discussionmentioning

confidence: 99%

“…The in vivo determination of the lifespan of RET in blood was determined by studying radioactive iron turnover (5,25). The existing methods of determination of RNA degradation rate involve direct determination of total RNA in a in vitro synchronized population of RET (6,8). One is able to determine the age structure of RET and RNA degradation rate in blood using the approach proposed in this work by FIG.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro culture data from rabbits show a linear decline of the RNA content at a rate of about 4% per hour that has been determined over a limited time (8). In rats, an exponential decline with half-life about 24 h was found (9).…”

mentioning

confidence: 99%

“…However, to fully understand aging of RET, the bone marrow RET have to be studied in a similar way. This would give a full picture of RET aging during homeostasis and under various nonhomeostatic conditions.Presented in the literature, methods for studying RET physiology often involve a difficult to perform isolation and an in vitro measurement of maturation of a synchronized population using radioactive tracers (5,6,8,25). The in vivo determination of the lifespan of RET in blood was determined by studying radioactive iron turnover (5,25).…”

mentioning

confidence: 99%

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Method of determination of the reticulocyte age distribution from flow cytometry count by a structured‐population model

Wiczling

Krzyzanski

2007

Cytometry Pt A

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Background: A flow cytometry count of reticulocytes (RET) provides information about distribution of signal emitted by reticulocyte RNA. A new method of determination of the RNA degradation rate in RET is provided. This technique allows one to determinate the age distribution of RET. Methods: The method is based on a series of flow cytometry counts of cultured RET. From those counts, the changes of signal distribution of RET over time are obtained. The RNA degradation rate of individual RET is then resolved based on changes in the signal distribution of the whole population of cells. The obtained relation between signal and age allows obtaining a RET age density that can be used to characterize RET age distribution and age dependence of processes that control the population dynamics. Results: The total maturation time of RET in rats is 3 days. The median time that a homeostatic RET spends in blood or before it becomes a mature RBC is about 0.6 days, whereas a stress RET needs 0.8 days. Conclusions:The proposed method provides means for studies in vivo RET dynamics using age-structured models. Key terms: reticulocytes; structured population model; RNA degradation; flow cytometry Reticulocytes (RET) are immature red blood cells containing one or more filaments or granules of ribosomal RNA. They are formed during mammalian erythropoiesis through the nuclear exclusion from last stage erythroblasts. Then, after a period of maturation in bone marrow, RET are released into the blood circulation and undergo further differentiation to mature erythrocytes. During that period, RET change from large and motile cells to small, biconcave, discoid cells (1-3). In rats, the RET residence time in the bone marrow is about 17-24 h (4,5). The in vivo disappearance of RET population from blood is linear and takes 30 h (5). In vitro culture studies showed that the RET lifespan in rats is about 2-3 days (5-7). The appearance of young RET in blood, called stress RET, gives an insight into the response of bone marrow to endogenous erythropoietin stimulation, treatment with recombinant human erythropoietin, or other erythropoietic stimuli. Hence, it is used as a relatively fast marker of undergoing erythropoiesis (2). The time stress RET spent in bone marrow can decrease to 6.5 h in rats.The lack of capacity for RNA synthesis in RET causes a progressive decrease in cell content of RNA and is one of the characteristics of maturation of RET and consequently provides means of assessing the RET maturation time. In vitro culture data from rabbits show a linear decline of the RNA content at a rate of about 4% per hour that has been determined over a limited time (8). In rats, an exponential decline with half-life about 24 h was found (9). An analogous decline with half-life of 43 h was reported in humans (10).Enumeration of peripheral RET is primarily done by means of automated counting. For flow cytometry measurements, the RET RNA is labeled with a fluorescence dye and the fluorescence of the dye-RNA complex is recorded (11). In general, youn...

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Section: Discussioncontrasting

confidence: 98%