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1.
Histochemical studies have been conducted by applying hexokinase (HK), aldolase (AD), glyceraldehyde-3-phosphate dehydrogenase (G3), succinate dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G6PD), and thiamine pyrophosphatase (TPPase) methods, as well as Nissl staining and Gomori's chrome-alum-hematoxylin-phloxine (CHP) methods to intercalated neurons of the supraoptic nucleus (SO) on Wistar strain rats. Intercalated neurons reacted weakly to the AD, G3, G6PD, and SDH tests, indicating that they belong to the category of ordinary neurons with low carbohydrate metabolism. Many fibrous astrocytes showing strong HK reactions surround neurosecretory neurons. However, they do not surround intercalated neurons with mild HK activity. These results indicate that the latter receive a poor supply of energy from glucose in the circulating blood in contrast to the former. Intercalated neurons are very rich in Nissl substance but lack CHP-positive material. They may have a high potential for synthesizing protein. The principal morphological features of the TPPase-positive Golgi material are peculiar and heterogeneous shape and poor development. These findings together with mild G6PD activity suggest that intercalated neurons are very likely to have poor synthesizing activity. 相似文献
2.
Prof. Dr. Koichi Iijima 《Histochemistry and cell biology》1971,25(2):107-122
Summary Detailed histochemical studies have been conducted on the distribution of thiamine pyrophosphatase (TPPase), hexokinase and glucokinase (HK), L-gulonolactone oxidase (GO), D-xylulose reductase (DX), L-xylulose reductase (LX) and ascorbic acid (AC) in every component of the locus coeruleus (LC) of the healthy adult male rabbit.The LC consisted of medium-sized neurons and small neurons. Both types of neurons were classified into the same five categories on the basis of the morphology of the Golgi apparatus (GA). Many intermediate forms were observed between these different categories. The present results concerning TPPase may indicate that each type of neuron goes through cyclic activity.The GA of the small neurons showed little variation in its reactivity and volume in each category and no disintegration or budding-off. These neurons were mildly positive for the HK test, and negative for the GO, DX, LX and AC tests in contrast to the medium-sized neurons. These results may suggest that the small neurons are metabolically inactive, and that they have a different function from the medium-sized neurons.The morphology of the GA of the medium-sized neurons was basically similar to that described for motor neurons. It was considerably different from the morphology of the GA reported in the dorsal vagal nucleus (X) and hypothalamic neurosecretory nuclei (HMN) of the rabbit. These results suggest that the medium-sized neurons of the LC may be motor neurons, and that they may not have a neurosecretory function.The medium-sized neurons showed strong activity whereas the surrounding glial cells and neuropil exhibited mild activity in the HK test. These findings may suggest that these neurons get their energy source directly from the circulating blood.The medium-sized neurons were mildly to moderately positive for the DX and LX tests, and some of them were strongly positive for the GO test. Positive granules showed the tendency to accumulate in a proximal part of the main cell process and the part of perikaryon adjacent to it for the AC test. On the basis of these results, it is suggested that there is a strong possibility that at least some of the medium-sized neurons of the LC have the ability to synthesize vitamin C. This ability may be intimately related to the ontogenetical development of catecholamine. 相似文献
3.
Summary A detailed histochemical study has been made on the mesenteric ganglia of the cat, and dorsal root ganglia of the squirrel monkey by the use of appropriate histochemical techniques accompanied by appropriate controls for phosphatases, esterases, and oxidative enzymes. The different neurons of a particular ganglion show varied amounts of enzyme activity at a particular time depending upon the functional state of the neurons. SDH, CYO and LDH reaction is prominent in the cytoplasm of the neurons, gliocytes and satellite cells, whereas the MAO preparations generally show a weak reaction. The AK is prominent in the neuropil, cell membranes and peripheral part of cytoplasm, whereas ATPase activity has been observed in blood vessels as well. In AC preparations the area of lipofuscin concentration shows more intense reaction than the rest of the cytoplasm. The activity of AChE and BChE varies from mild, to moderate to strong. The TPPase preparations show morphologically different types and amounts of TPPase positive Golgi material even in the adjoining cells. The relationship between the TPPase Golgi material and various oxidative and dephosphorylating enzymes has been briefly discussed.Abbreviations used AC
Acid phosphatase
- AChE
Acetyl-cholinesterase (specific)
- AK
Alkaline phosphatase
- AMPase
Adenosine monophosphatase (5-nucleotidase)
- ATPase
Adenosine triphosphatase
- BChE
Butyryl-cholinesterase (nonspecific)
- CYO
Cytochrome oxidase
- DPN-D
DPN-diaphorase
- G6P
Glucose-6-phosphatase
- LDH
Lactic dehydrogenase
- MAO
Monoamine oxidase
- MDH
Malic dehydrogenase
- NAD-D
NAD-diaphorase
- SDH
Succinic dehydrogenase
- SE
Simple esterase
- TPPase
thiamine pyrophosphatase
T. R. Shanthaveerappa in previous publications. 相似文献
4.
Summary Detailed histochemical studies have been conducted on the distribution of various enzymes such as thiamine pyrophosphatase,
α-glucan phosphorylase, hexokinase, glucose-6-phosphate dehydrogenase, aldolase, lactate dehydrogenase and succinate dehydrogenase
in various components of the nucleusEdinger-Westphali, nucleus n. oculomotorii, nucleus ruber and nucleus niger of healthy adult male Wistar strain rats.
The thiamine pyrophosphatase reaction showed the morphological patterns of the Golgi apparatus characteristic for each nucleus.
The Golgi apparatus was well developed in the nucleusEdinger-Westphali, composing a network of highly fenestrated plates in the nucleus n. oculomotorii and nucleus ruber, and a simple network
in the nucleus niger. These results indicate that the former three nuclei need a rich energy supply and argue against the
possibility that the four nuclei have a secretory role.
The neurons of the nucleusEdinger-Westphali may derive their energy mainly from glucose of the circulating blood, but glial cells may serve as energy donators to the
neurons in the pars compacta of the nucleus niger, and the neurons of the other nuclei may derive energy from both sources.
These conclusions are consistent with the morphological patterns of the Golgi apparatus.
It is suggested that the neurons of the nucleusEdinger-Westphali, nucleus n. oculomotorii, nucleus ruber and of the pars lateralis of the nucleus niger may be equipped almost equally with
the Embden-Meyerhof pathway and with the hexose monophosphate shunt. But, the hexose monophosphate shunt is dominant in the
pars compacta of the nucleus niger. It is also suggested that the pattern of distribution of succinate dehydrogenase may parallel
that of lactate dehydrogenase. The nucleus n. oculomotorii, and nucleus ruber have a higher level of oxidative metabolism
than the nucleusEdinger-Westphali and the nucleus niger. The nucleusEdinger-Westphali may be representative of autonomic nuclei with low oxidative metabolism whereas the nucleus n. oculomotorii may represent
motor nuclei with high oxidative metabolism. Predominance of hexose monophosphate shunt, intense hexokinase reaction around
the neurons, and weak activity of succinate dehydrogenase indicate that the pars compacta of the nucleus niger belongs to
the category of “exceptional nuclei”. 相似文献
5.
Summary Detailed histochemical studies have been performed on the morphology of the Golgi apparatus (GA) by application of the thiamine pyrophosphatase (TPPase) method (Novikoff and Goldfischer, 1961) to the neurons of the locus coeruleus (LC) of normal and catecholamine biosynthesis inhibitors (fusaric acid and D, L--methyl-p-tyrosine methylester HCl) given adult healthy male Wistar strain rats. The neurons were classified into five categories on the basis of the morphology of the Golgi apparatus. The number of cells in individual categories was counted to evaluate the percentage of each category in the whole nucleus.The majority of cells belongs to Types II, III, and IV whose GA goes through cyclic activity, but the remaining neurons belong to Types I and V which may have a strong tendency to be different from the former in character. The latter neurons correspond formally with Types I and V of the rabbit LC, but they do not respond to the drugs administered. The rat LC is very similar to the dorsal vagal nucleus of the rabbit in regard to the dominant category. The present results indicate that the majority of the rat LC neurons may work vigorously and they may be motor neurons.Administration of the drugs caused reduction of TPPase activity, augmentation of disintegration and the budding-off process of the GA of Type IV, a decrease in the percentage of Type IV and an increase in that of Type II. Administration of 100 mg/kg fusaric acid caused maximal morphological change of the GA at the 90th minute; however, administration of 200 mg/kg fusaric acid showed more marked change of the GA, having two peaks and two valleys. The GA revealed much more intense reaction to D,L--methyl-p-tyrosine methylester HCl than to fusaric acid. The present results indicate that tyrosine hydroxylase may be the rate-limiting enzyme in the catecholamine biosynthesis.These noticeable changes of GA caused by administration of the drugs were completely restricted to the neurons of LC and the neurons of the mesencephalic nucleus of the trigeminal nerve did not show any morphological changes of the GA. These results strongly suggest that the GA of the rat LC neurons may have ability to synthesize catecholamine whereas the GA of the rat mesencephalic nucleus of the trigeminal nerve may be completely devoid of this ability and that the role of the GA may be different depending on the anatomical regions. 相似文献
6.
Badia Bisbis Frédérik le Dily Claire Kevers Jean-Pierre Billard Claude Huault Thomas Gaspar 《Plant Growth Regulation》1993,13(3):257-261
Habituated (H) nonorganogenic sugarbeet callus was found to exhibit a disturbed sugar metabolism. In contrast to cells from normal (N) callus, H cells accumulate glucose and fructose and show an abnormal high fructose/glucose ratio. Moreover, H cells which have decreased wall components, display lower glycolytic enzyme activities (hexose phosphate isomerase and phosphofructokinase) which is compensated by higher activities of the enzymes of the hexose monophosphate pathway (glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase). The disturbed sugar metabolism of the H callus is discussed in relation to a deficiency in H2O2 detoxifying systems.Abbreviations 6PG-DH
6-phosphogluconate dehydrogenase
- G6P-DH
glucose-6-phosphate dehydrogenase
- H
fully habituated callus
- HK
hexokinase
- HMP
hexoses monophosphate
- HPI
hexose phosphate isomerase
- N
normal callus
- PFK
phosphofructokinase 相似文献
7.
Human breast cancer cell lines have been shown to possess high affinity receptors for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and their growth is inhibited by this steroid. The present study examines the effect of 1,25(OH)2D3 on the activity of glucose-6-phosphate dehydrogenase (G6PD) in cells of a human breast cancer cell line MCF-7. G6PD, an enzyme which controls the hexose monophosphate shunt, is elevated and sensitive to 17 beta-estradiol in breast tumors. G6PD activity was stimulated by 1,25(OH)2D3 in a dose-dependent manner at very low concentrations of steroid (10(-10)-10(-12) M). 1,25(OH)2D3 increased maximum velocity without modifying the affinity constant of the enzyme for glucose-6-phosphate. 相似文献
8.
X linkage of phosphoglycerate kinase in the mouse 总被引:10,自引:0,他引:10
The levels of phosphoglycerate kinase (PGK), glucose 6-phosphate dehydrogenase (G6PD), and lactate dehydrogenase (LDH) were measured in one-cell embryos from X/0 and X/X females. Since the level of both PGK and G6PD was dependent on the number of X chromosomes present in the mother, these two enzymes are most likely coded for by X-linked genes. The level of LDH was the same in both types of embryos, indicating autosomal linkage. A search for an electrophoretic variant of PGK was not successful. 相似文献
9.
D. R. Joanisse K. B. Storey 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1994,164(3):247-255
The activity of some enzymes of intermediary metabolism, including enzymes of glycolysis, the hexose monophosphate shunt, and polyol cryoprotectant synthesis, were measured in freeze-tolerant Eurosta solidaginis larvae over a winter season and upon entry into pupation. Flexible metabolic rearrangement was observed concurrently with acclimatization and development. Profiles of enzyme activities related to the metabolism of the cryoprotectant glycerol indicated that fall biosynthesis may occur from two possible pathways: 1. glyceraldehyde-phosphate glyceraldehyde glycerol, using glyceraldehyde phosphatase and NADPH-linked polyol dehydrogenase, or 2. dihydroxyacetonephosphate glycerol-3-phosphate glycerol, using glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase. Clearance of glycerol in the spring appeared to occur by a novel route through the action of polyol dehydrogenase and glyceraldehyde kinase. Profiles of enzyme activities associated with sorbitol metabolism suggested that this polyol cryoprotectant was synthesized from glucose-6-phosphate through the action of glucose-6-phosphatase and NADPH-linked polyol dehydrogenase. Removal of sorbitol in the spring appeared to occur through the action of sorbitol dehydrogenase and hexokinase. Glycogen phosphorylase activation ensured the required flow of carbon into the synthesis of both glycerol and sorbitol. Little change was seen in the activity of glycolytic or hexose monophosphate shunt enzymes over the winter. Increased activity of the -glycerophosphate shuttle in the spring, indicated by greatly increased glycerol-3-phosphate dehydrogenase activity, may be key to removal and oxidation of reducing equivalents generated from polyol cryoprotectan catabolism.Abbreviations 6PGDH
6-Phosphogluconate dehydrogenase
- DHAP
dihydroxy acetone phosphate
- F6P
fructose-6-phosphate
- F6Pase
fructose-6-phospha-tase
- FBPase
fructose-bisphosphatase
- G3P
glycerol-3-phosphate
- G3Pase
glycerol-3-phosphate phophatase
- G3PDH
glycerol-3-phosphate dehydrogenase
- G6P
glucose-6-phosphate
- G6Pase
glucose-6-phosphatase
- G6PDH
glucose-6-phosphate dehydrogenase
- GAK
glyceraldehyde kinase
- GAP
glyceraldehyde-3-phosphate
- GAPase
glyceraldehyde-3-phosphatase
- GAPDH
glyceraldehyde-3-phosphate dehydrogenase
- GDH
glycerol dehydrogenase
- GPase
glycogen phosphorylase
- HMS
hexose monophosphate shunt
- LDH
lactate dehydrogenase
- NADP-IDH
NADP+-dependent isocitrate dehydrogenase
- PDHald
polyol dehydrogenase, glyceraldehyde activity
- PDHgluc
polyol dehydrogenase, glucose activity
- PFK
phosphofructokinase
- PGI
phosphoglucoisomerase
- PGK
phosphoglycerate kinase
- PGM
phosphoglucomutase
- PK
pyruvate kinase
- PMSF
phenylmethylsulfonylfluoride
- SoDH
sorbitol dehydrogenase
-
V
max
maximal enzyme activity
- ww
wet weight 相似文献
10.
C. Junien H. Rubinson J. C. Dreyfus M. C. Meienhofer N. Ravise J. Boué A. Boué 《Human genetics》1976,33(1):61-66
Summary The activity of 13 cytoplasmic enzymes has been determined in fibroblast extracts from 9 triploid and 13 control lines. The results show a high activity for 2 X-linked enzymes, glucose 6-phosphate dehydrogenase and phosphoglycerate kinase. These data, together with cytogenetic observations, support the contention that 2 X chromosomes were active in the triploid lines.Abbreviations G6PD
Glucose 6-phosphate dehydrogenase
- 6PGD
6-phosphogluconate dehydrogenase
- HK
hexokinase
- PGM
phosphoglucomutase
- PHI
phosphohexoisomerase
- PFK
phosphofructokinase
- ALD
aldolase
- TPI
triosephosphate isomerase
- PGK
phosphoglycerate kinase
- ENOL
enolase
- AK
adenylate kinase
- LDH
lactic dehydrogenase
- HBDH
hydroxybutyrate dehydrogenase
INSERM U. 129.INSERM U. 73. 相似文献
11.
Mohammad Jodeiri Farshbaf 《生物学前沿》2017,12(3):175-182
Background
The prevalence of neurodegenerative disorders such as Parkinson’s disease (PD) is increased by age. Alleviation of their symptoms and protection of normal neurons against degeneration are the main aspects of the researches to establish novel therapeutic strategies. Many studies have shown that mitochondria as the most important organelles in the brain which show impairment in PD models. Succinate dehydrogenase (SDH) as a component of the oxidative phosphorylation system in mitochondria connects Krebs cycle to the electron transport chain. Dysfunction or inhibition of the SDH can trigger mitochondrial impairment and disruption in ATP generation. Excessive in lipid synthesis and induction of the excitotoxicity as inducers in PD are controlled by SDH activity directly and indirectly. On the other hand, mutation in subunits of the SDH correlates with the onset of neurodegenerative disorders. Therefore, SDH could behave as one of the main regulators in neuroprotection.Objective
In this review we will consider contribution of the SDH and its related mechanisms in PD.Methods
Pubmed search engine was used to find published studies from 1977 to 2016. “Succinate dehydrogenase”, “lipid and brain”, “mitochondria and Parkinson’s disease” were the main keywords for searching in the engine.Results
Wide ranges of studies (59 articles) in neurodegenerative disorders especially Parkinson’s disease like genetics of the Parkinson’s disease, effects of the mutant SDH on cell activity and physiology and lipid alteration in neurodegenerative disorders have been used in this review.Conclusion
Mitochondria as key organelles in the energy generation plays crucial roles in PD. ETC complex in this organelle consists four complexes which alteration in their activities cause ROS generation and ATP depletion. Most of complexes are encoded by mtDNA while complex II is the only part of the ETC which is encoded by nuclear genome. So, focusing on the SDH and related pathways which have important role in neuronal survival and SDH has a potential to further studies as a novel neuroprotective agent.12.
In order to study the mechanism of X Chromosome (Chr) inactivation in marsupials, the cDNA for glucose-6-phosphate dehydrogenase (G6PD) from an Australian marsupial, the wallaroo (Macropus robustus), was cloned. A partial clone containing the 3 half of the cDNA was obtained by screening a liver cDNA library. The majority of the coding region was obtained by polymerase chain reaction of cDNA with primers designed from regions of conservation between human and opossum G6PD. The 5 end was obtained by rapid amplification of cDNA ends. High homology was observed between mammalian species in the coding region. The 5 untraslated region is highly G+C rich, and appears to be part of a CpG island, as is the case in the human and mouse genes. This is the first report of the full sequence of the cDNA for any marsupial X-linked gene.The nucleotide sequence data reported in this paper have been submitted to GenBank and assigned accession number U13899. 相似文献
13.
Summary We produced somatic cell hybrids between HT 1080-6TG human fibrosarcoma cells and either rat white blood cells (WBC) or cells directly derived from rat spleen. Karyologic and isozyme analyses of hybrid cells indicated that they preferentially lose rat chromosomes. Hypoxanthine-aminopterine thymidine-selected hybrid clones expressing rat hypoxanthine phosphoribosyltransferase (HPRT), glucose-6-phosphate dehydrogenase (G6PD), and phosphoglycerate kinase (PGK) and containing the rat X chromosome were counterselected in a medium containing 30 g/ml of 6-thioguanine. Concordant loss of the rat X chromosome and of the expression of rat HPRT and G6PD was observed in the hybrid clones. 相似文献
14.
Ultrastructural localization of phosphatase activity in the guinea pig pineal gland by the cerium technique 总被引:1,自引:0,他引:1
Thiamine pyrophosphatase (TPPase), nucleoside diphosphatase (NDPase), and glucose-6-phosphatase (G-6-Pase) were localized by the cerium technique in guinea pig pinealocytes and compared with the corresponding lead technique. NDPase and TPPase were also compared at different pH values using the cerium technique. Vibratome sections of perfusion-fixed tissue were incubated with cerium chloride or lead nitrate. Substrates used were thiamine pyrophosphate (for TPPase), sodium inosine diphosphate (NDPase), and disodium glucose-6-phosphate (G-6-Pase). The 1-2 trans saccules of the Golgi apparatus showed TPPase and NDPase activity but none for G-6-Pase. The endoplasmic reticulum (ER) cisternae and perinuclear space had NDPase and G-6-Pase activity but not TPPase. The abluminal plasmalemma of endothelial cells and the plasmalemma of Schwann cells demonstrated TPPase and NDPase activity but the luminal plasmalemma of the endothelial cells and the plasmalemma of pinealocyte processes showed only NDPase activity. TPPase was active at all pH values tested, but NDPase was most active at pH values of 6.5 and 7.0. Lead phosphate precipitate was frequently seen in nuclei, perinuclear space, ER cisternae, and "synaptic" vesicles when lead was used as the capturing agent. These sites were usually not labeled when cerium was used. 相似文献
15.
Paolino Ninfali Luciano Baronciani Annamaria Ruzzo Cinzia Fortini Elisa Amadori Gionni Dall'ara Mauro Magnani Ernest Beutler 《Human genetics》1993,92(2):139-142
In the Ferrara district, an area south of the Po delta, four different variants of glucose-6-phosphate dehydrogenase (G6PD;E.C.1.1.49) have been described as a result of biochemical characterization of the enzyme protein: one was G6PD Mediterranean (G6PD Med) and three were local variants named Ferrara I, II, and III. The Ferrara I variant was recently analysed at the DNA level and shown to correspond to G6PD A376G/202A, while the mutations causing the variants II and III, still remain unknown. We analysed the G6PD coding region of 18 apparently unrelated G6PD deficient subjects, whose families have lived in the Ferrara district for at least three generations: 12 subjects had G6PD Med563T/1311T, 3, G6PD Santamaria376G/542T and 2, G6PD A-376G/202A. In one subject we found a new mutation, a GA transition at nucleotide 242 causing an ArgHis amino acid replacement at position 81. We named this new variant G6PD Lagosanto242 A. Phenotypically the enzyme has nearly normal kinetic properties and appears different from the variants Ferrara II and III. 相似文献
16.
Rapid release of bound glucose-6-phosphate dehydrogenase by growth factors. Correlation with increased enzymatic activity 总被引:3,自引:0,他引:3
R C Stanton J L Seifter D C Boxer E Zimmerman L C Cantley 《The Journal of biological chemistry》1991,266(19):12442-12448
Epidermal growth factor (EGF), a mitogen for renal proximal tubule cells, activated the hexose monophosphate (HMP) shunt in renal proximal tubule cells (Stanton, R. C., and Seifter, J. L. (1988) Am. J. Physiol. 254, C267-C271). We therefore evaluated the effect of EGF on the HMP shunt enzymes glucose 6-phosphate dehydrogenase (G6PD, the rate-limiting enzyme) and 6-phosphogluconate dehydrogenase. Rat renal cortical cells (RCC) were incubated with either EGF or platelet-derived growth factor (PDGF) and then assayed for G6PD and 6-phosphogluconate dehydrogenase activities. EGF and PDGF increased G6PD activity by 25 and 27% respectively. Although phorbol myristate acetate (PMA), ionomycin, PMA + ionomycin, and 8-bromo-cyclic AMP had no significant effect on the activity, a 5-min preincubation with PMA potentiated the activation of G6PD by PDGF. Growth factor activation of G6PD was also seen in a fibroblast and epithelial cell line. None of the agents affected 6-phosphogluconate dehydrogenase activity in the RCC or in the cell lines. Further exploration into a possible mechanism for G6PD activation revealed that growth factors caused release of G6PD from a structural element within the cell. Streptolysin O permeabilization of RCC did not cause significant release of G6PD. However, within 1 min of addition of EGF or PDGF to permeabilized cells, G6PD was released into the cell supernatant. The nonhydrolyzable analog of GTP, guanosine 5'-O-(thiotriphosphate), caused a similar release of G6PD. Preincubation with pertussis toxin or guanyl-5'-yl thiophosphate inhibited the PDGF but not the EGF effect. Although the data do not establish a definitive proof linking G6PD release and G6PD activation, these results suggest that they are related. Thus, growth factor stimulation of the HMP shunt likely occurs by a novel mechanism associated with release of bound G6PD. 相似文献
17.
F. De Graaf W. Van Raamsdonk E. Van Asselt P. C. Diegenbach 《The Histochemical journal》1991,23(6):273-280
Summary Enzyme histochemical profiles of spinal motoneurons in the zebrafish were determined. Five enzymes of glucose metabolism were chosen: glucose-6-phosphate dehydrogenase (G6PDH), hexokinase (HK), phosphofructokinase (PFK), succinate dehydrogenase (SDH) and NADH tetrazolium reductase (NADH-TR). Motoneurons were traced with Fluorogold and classified as those that innervate white muscle fibres (W-MNs) and those that innervate red and intermediate muscle fibres (R/ I-MNs). The average enzyme activities per volume of tissue in the somata of both populations differed at most by 25%. Both the average soma volume and the average number of muscle fibres innervated are three times larger for the W-MNs than for the a/I-MNs. This suggests that the total amount of enzyme activity within a neuron soma matches target size.In the R/I-MNs, the activities of SDH and NADH-TR were closely correlated (correlation coefficient, r=0.99;p<0.05) and HK activity correlated well with G6PDH activity (r=0.94;p<0.05), butnot with PFK (r=0.64;p>0.05). In the W-MNs, there was no correlation between SDH and NADH-TR (r=–0.59;p>0.05) or between HK and G6PDH (r=0.50;p>0.05) and the correlation coefficient between HK and PFK activity was close to zero (r=0.04;p>0.05).It was concluded that in the R/I-MNs gwhich are continuously ctive, firing activity is fuelled by oxidative metabolsm. We suggest that in the W-MNs glucose is stored in the form of glycogen and that, despite high levels of NADH-TR present, the energy for intermittent firing activity is provided by glycolysis. 相似文献
18.
Substrate specificity and product inhibition of different forms of fructokinases and hexokinases in developing potato tubers 总被引:10,自引:0,他引:10
The substrate dependence and product inhibition of three different fructokinases and three different hexokinases from growing potato (Solanum tuberosum L.) tubers was investigated. The tubers contained three specific fructokinases (FK1, FK2, FK3) which had a high affinity for fructose K
m=64, 90 and 100 (M) and effectively no activity with glucose or other hexose sugars. The affinity for ATP (K
m=26, 25 and 240 M) was at least tenfold higher than for other nucleoside triphosphates. All three fructokinases showed product inhibition by high fructose (K
i=5.7, 6.0 and 21 mM) and were also inhibited by ADP competitively to ATP. Sensitivity to ADP was increased in the presence of high fructose, or fructose-6-phosphate. In certain conditions, the K
i (ADP) was about threefold below the K
m (ATP). All three fructokinase were also inhibited by fructose-6-phosphate acting non-competitively to fructose (K
i=1.3 mM for FK2). FK1 and FK2 showed very similar kinetic properties whereas FK3, which is only present at low activities in the tuber but high activities in the leaf, had a generally lower affinity for ATP, and lower sensitivity to inhibition by ADP and fructose. The tuber also contained three hexokinases (HK1, HK2, HK3) which had a high affinity for glucose (K
m=41, 130 and 35 M) and mannose but a poor affinity for fructose (K
m=11, 22 and 9 mM). All three hexokinases had a tenfold higher affinity for ATP (K
m=90, 280 and 560 M) than for other nucleoside triphosphates. HK1 and HK2 were both inhibited by ADP (K
i=40 and 108 M) acting competitively to ATP. HK1, but not HK2, was inhibited by glucose-6-phosphate, which acted non-competitively to glucose (K
i=4.1 mM). HK1 and HK2 differed, in that HK1 had a narrower pH optimum, a higher affinity for its substrate, and showed inhibition by glucose-6-phosphate. The relevance of these properties for the regulation of hexose metabolism in vivo is discussed.Abbreviations FK
fructokinase
- Fru6P
fructose-6-phosphate
- Glc6P
glucose-6-phosphate
- HK
hexokinase
- NTP
nucleoside triphosphate
- Pi
inorganic phosphate
- UDPGlc
uridine-5-diphosphoglucose
This work was supported by the Deutsche Froschungsgemeinschaft (SFB 137). We are grateful to Professor E. Beck (Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, FRG) for providing laboratory facilities. 相似文献
19.
Stephen R. Palumbi Bruce D. Sidell Rebecca Van Beneden Glenn D. Smith Dennis A. Powers 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1980,138(1):49-57
Summary In order to evaluate the role of glucose-phosphate isomerase (GPI) inFundulus heteroclitus, the isozymes and allozymes were purified and some of their physical and kinetic properties determined.Isozymes were purified from both liver (GPI-B) and muscle (GPI-A) tissue (Tables 1, 2). Gel filtration of the native enzyme and SDS-polyacrylamide gel electrophoresis indicated that all forms are dimers of approximately 110,000 Daltons (Figs. 4, 5). Although thermal stability studies revealed no differences between the allozymes, the isozymes were clearly different (Figs. 6, 7). Kinetic analysis showed further differences between isozymes inK
m for substrate andK
I for 6-phosphogluconate (Figs. 8, 9; Table 3). No significant differences were found between the allozymes of the B-locus under the conditions employed in this study.Based on the tissue specificities and the functional differences between isozymes, we propose a possible regulatory role for GPI-B inF. heteroclitus. The sensitivity of this isozyme to 6-phosphogluconate inhibition may allow GPI-B to act as a regulatory enzyme in the partitioning of carbon flow between glycolysis and the hexose monophosphate shunt.Abbreviations me
-mercaptoethanol
-
F6P
fructose-6-phosphate
-
G1P
glucose-1-phosphate
-
G6P
glucose-6-phosphate
-
G6Pase
glucose-6-phosphatase
-
G6PDH
glucose-6-phosphate dehydrogenase
-
GPI
glucosephosphate isomerase
-
HK
hexokinase
-
HMP
hexose monophosphate shunt
-
6PG
6-phosphogluconate
-
PGM
phosphoglucomutase
Supported in part by: NSF grants DEB-76-19877 to D.A.P. and PCM 77-16838 to B.D.S., NIH Biomedical grant 5-50-7RR07-041 and a grant from the National Geographic Society. G.D.S. and R.V.B. are NIH trainees supported by a training grant (No. HD00139) to the Department of Biology, The Johns Hopkins University. This is contribution No. 1052 from the Department of Biology 相似文献
20.
Masaaki Satoh Hiroichi Asagami Dongchon Kang Shigeki Minakami Koichiro Takeshige 《Molecular and cellular biochemistry》1995,152(2):159-165
A chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (fMLP), induced an acidification of cytosol by about 0.05 pH units in 30 sec followed by an alkalinization in human neutrophils. The quantitative contribution of acid production to the acidification was studied. The superoxide (O2
–) production stimulated by fMLP was not involved in the acidification because the production of acids in neutrophils from patients with chronic granulomatous disease who do not produce O2
–, was the same as that in normal neutrophils. The intracellular acidification was completely inhibited by deoxyglucose, suggesting that energy metabolism enhanced upon stimulation by fMLP might be the main source of the acidification. Although enhancement of the lactate formation by fMLP was 0.8 nmol/106 cells, which could lower intracellular pH by 0.08 pH units, the lactate production could not explain the initial acidification because the production of lactate started at 1 min after the stimulation while the intracellular acidification began immediately after the stimulation. Mitochondrial respiratory inhibitors such as KCN and rotenone had no effects on the fMLP-induced intracellular acidification. The fMLP-induced production of CO2 in 30 sec through the hexose monophosphate shunt was only 2.6 pmol/106 cells, which was calculated to decrease intracellular pH by only 0.0014. Thus, changes of energy metabolism induced by fMLP does not explain the acidification.Abbreviations fMLP
N-formyl-methionyl-leucyl-phenylalanine
- BCECF-AM
2,7-bis(carboxyethyl)carboxyfluorescein acetoxymethyl ester
- PMA
phorbol 12-myristate 13-acetate
- CGD
chronic granulomatous disease
- HMP
hexose monophosphate
- pHi
intracellular pH 相似文献