Changes in activities and isozyme patterns of glycolytic enzymes during erythroid differentiation in vitro.
نویسندگان
چکیده
Late committed progenitor cells of erythropoiesis, CFU-E (colony-forming unit--erythroid), were isolated from mouse spleens to near homogeneity by a three-step enrichment procedure. The procedure included a four-day pretreatment of bled mice with the antibiotic thiamphenicol, a recovery period of 3 1/2 days, followed by centrifugal elutriation and Percoll density gradient centrifugation of the spleen cells. This practically pure CFU-E population was used to study some aspects of erythroid differentiation in vitro. Colony growth, as well as morphology and glycolytic enzyme activities of cells isolated at selected times of the 48-hour culture period, were determined. Marked declining activities of several enzymes, including hexokinase, phosphofructokinase, aldolase, enolase, pyruvate kinase, and glucose-6-phosphate dehydrogenase, were observed during in vitro differentiation. The activity of diphosphoglycerate mutase was almost absent in the CFU-E, but progressively increased during differentiation. The isozyme distribution of aldolase and enolase did not change during CFU-E in vitro differentiation into the reticulocyte. Hexokinase (HK) in the CFU-E contained mainly a double-banded type I isozyme, in addition to a minor amount of HK II. During differentiation, a shift was noticed within the double-banded HK I region, whereas HK ii disappeared after one cell division. Pyruvate kinase in the CFU-E was characterized by the presence of both the K-type and the L-type isozyme and hybrids of these isozyme types. During in vitro differentiation, the production of the K-type isozyme rapidly stops in favor of the L type.
منابع مشابه
PECTIC ENZYME PATTERNS OF FUSARIUM OXYSPORUM VIRULENT ISOLATES FROM CHICKPEA IN IRAN
The pectic enzymes produced in vitro by 8 isolates (5 Highly virulent and 3 Weakly virulent) of Fusarium oxysporum , were detected by spectrophotometry, and characterized by polyacrylamide gel electrophoresis with substrate-containing gels (zymogram). Analysis of the polygalacturonase (PG) isozyme banding patterns (zymogram) identified two distinct phenotypes among the isolates from chickpea (C...
متن کاملEvaluation of Antioxidant Enzyme Activities in Withania somnifera L. in vitro and in vivo Grown Explants
Background: Withania somnifera L. Dunal, (Ashwagandha) belongs to the family Solanaceae and is known as ‘Indian ginseng’ in the traditional system of medicine. Objectives: In the present investigation, comparative analysis was carried out between two different ecotypes of W. somnifera related to total soluble protein and antioxidant enzyme activity [catalase (CAT), s...
متن کاملDifferentiation of Mouse Yolk Sac Cells to Erythroid Cells in The Presence of Erythropoietin
Purpose: Yolk sac hematopoietic stem cells (YS-HSC) have two dominant characteristices: a larger reproductive capacity and the absence of the expression of MHC associated antigens. Therefore, these cells are promising candidates for transplantation, cell therapy and gene manipulation. There are controversial reports on the effects of erythropoietin (EPO) on the differentiation of yolk sac cells...
متن کاملChanges of digestive enzymes activity in Caspian Kutum (Rutilus frisii kutum) during larval developmental stages
The ontogenesis and specific activities of pancreatic (trypsin, chymotrypsin, amylase and lipase) and intestinal enzymes (alkaline phosphatase, aminopeptidase N) were investigated in Kutum (Rutilus frisii Kutum) from the onset of exogenous feeding (3 day after hatching, DAH) to the juvenile stage at 50 DAH. Trypsin- and chymotrypsin-specific activity showed similar patterns and increased with l...
متن کاملAntioxidant Isoenzymes Activities in Seedling Roots of Wheat Exposed to Drought Stress
Abstract This study was conducted to evaluate the role of oxidative stress in roots of eight wheat genotypes with different drought tolerance, subjected to drought stress. For analyzing the changes of antioxidant enzymes, native PAGE analyses of protein extract were performed. The roots of wheat seedlings showed one unambiguous isoform of superoxide dismutase (SOD) and catalase (CAT). Eight iso...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Blood
دوره 64 3 شماره
صفحات -
تاریخ انتشار 1984