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101.
The effects of phorbol ester and forskolin on the net phosphorylation and turnover of P0 phosphate groups was studied in normal and exprimentally diabetic rats. In sciatic nerve segments isolated from normal rats and incubated with [32P]-inorganic phosphate, phosphorylation of the major peripheral myelin protein, P0, was increased 2–5 fold in a time and dose-dependent manner by phorbol 12,13 dibutyrate (PDB). This increase was blocked by the protein kinase inhibitors, H-7 and staurosporine. Both the basal and PDB-stimulated phosphorylation of P0 were significantly greater in segments of sciatic nerve from streptozotocin-induced diabetic rats. Prolonged exposure of nerve segments to PDB abolished the stimulated phosphorylation of P0 and immunoblots of nerve proteins revealed a decrease in the content of the protein kinase C -isoform. The adenylate cyclase activator, forskolin, had no affect on the PDB-stimulated phosphorylation of P0 in normal nerve but decreased phosphorylation in diabetic nerve. To measure turnover of P0 phosphate groups, nerves were incubated with32P and incorporated label was then chased in radioactivity-free medium for up to 4 hours. P0 from normal nerve prelabeled under basal conditions lost 25% of its radioactivity during this time. In contrast, nearly all of the additional phosphate groups prelabeled in the presence of PDB disappeared after 2 hours of chase. P0 phosphate groups from diabetic nerve displayed similar turnover kinetics. When forskolin was added to the chase medium, the turnover of P0 phosphate moieties was accelerated in normal, but not in diabetic nerve. These findings clearly establish a prominent role for protein kinase C in P0 phosphorylation, provide evidence for heterogeneous turnover of P0 phosphate groups and suggest that cyclic AMP-mediated processes may modulate P0 phosphorylation. Further, these results indicate that the metabolism of P0 phosphate moieties is perturbed in nerve from diabetic animals.Special issue dedicated to Dr. Marjoris B. Lees. 相似文献
102.
Vascular smooth muscle cell membranes from prehypertensive rats of the Milan hypertensive strain (MHS) were used to examine adenylyl cyclase activity and its regulation by guanine nucleotide regulatory proteins (G-proteins). Basal adenylyl cyclase activity was similar in MHS and Milan normontensive strain (MNS) membranes. Forsokolin (10?4 M) produced a significantly greater stimulatory response in MHS membranes, but this was not observed with NaF (10?2 M). Isoporterenol (10?4 M) caused a significantly decreased stimulation of adenylyl cyclase activity in MHS membranes, while prostaglandin E1 (10?5 M) produced similar responses in the two strains. Gi function and GTP responses, as observed by biphasic effects of GTP on isoproterenol-stimulated membranes, were similar in both strains. The levels of Gi2α and Gqα/G11α were similar in the two strains, while the levels of Gsα (44 and 42 kDa forms) and the β-subunit were significantly reduced by ~20% in MHS membranes. The α-subunit of Gi3 was dramatically reduced by ~80% in MHS membranes. The affinities of β-adrenergic receptors for the antagonist, cyanophindolol, were similar in the two strains; however, the number of β-adrenoceptors was substantially reduced in MHS membranes. These findings may be of relevance to altered vascular reactivity and transmembrane ion distribution observed in the MHS. 相似文献
103.
J. L. Albi J. Planas J. Sánchez 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1993,163(2):118-122
Glucose metabolism in peripheral blood lymphocytes from the brown trout Salmo trutta has been studied. Glucose is taken up by means of a sodium-independent saturable process (K
m=10.8 mmol·l-1), as well as by simple diffusion. Once within the cell, most of glucose is directed to lactate production through either the Embden-Meyerhof pathway or the hexose-monophosphate shunt. Rates of lactate formation are higher than rates of CO2 formation. Glutamine does not exert an effect on either glucose uptake or glucose metabolism. The present study provides information regarding the nature of energy sources for different cell types in salmonids.Abbreviations 3-OMG
3-O-methyl glucose
- EM
Embden-Meyerhoff pathway
- G6D
glucose-6-phosphate dehydrogenase
- HK
hexokinase
- HMS
hexose monophosphate shunt
- ICDH
isocitrate dehydrogenase
-
K
m
apparent Michaelis constant
- LDH
lactate dehydrogenase
- MCB
modified Cortland buffer
- PBL
peripheral blood lymphocytes
- PFK
fructose-6-phosphate kinase
- PK
pyruvate kinase
- RBC
red blood cells
-
V
max
maximal rate of uptake 相似文献
104.
105.
Elena Driomina Igor Polnikov Victor Sharov Ofelia Azizova Yury Vladimirov 《Free radical research》1994,20(5):279-288
A chemiluminescence (CL) flash kinetics on the addition of Fe2+ ions into oxidized low density lipoprotein (LDL) suspension has been studied. LDL oxidation was carried out at 37°C without and in the presence of 5 or 50 μM of Cu.2+ It has been found that under certain experimental conditions (the addition of excess iron ions, more than 1 mM) the amplitude of CL flash depended almost linearly (1) on the concentration of oxidized LDL and (2) on the extent of LDL oxidation measured as diene conjugates (DC) and 2-thiobarbituric acid-reactive substance (TBARS) accumulation. The corresponding correlation coefficients were: for TBARS - 0.94 and for DC - 0.97, in the case of LDL autooxidation; 0.72 and 0.98, in the case of copper-induced LDL oxidation. A sensitivity of the CL method was shown to be significantly enhanced (by more than two orders) in the presence of CL sensitizer - 2, 3,5, 6-lH,4H-tetrahydro-9-(2' -benzoimidazolyl)-quinolizin-(9, 9a, 1 -gh)coumarin. 相似文献
106.
Gino Giannaccini Claudia Martini Antonio Lucacchini Enrica Strettoi Marco Piccolino† 《Journal of neurochemistry》1993,61(4):1263-1269
Abstract: The binding of [3 H]flunitrazepam, [3 H]RO 5-4864, and [3 H]PK 11195 to membrane preparations of the retina was studied in the turtle and rabbit. Only a single population of [3 H]flunitrazepam binding sites was detected in the turtle, whereas two populations appeared to be present in the rabbit. No specific binding for [3 H]RO 5-4864 and [3 H]PK 11195 could be detected in the turtle. In rabbit, both ligands bound with high affinity, revealing a significant population of binding sites (KD values of 24 ± 2.3 and 2.2 ± 0.8 nM, and Bmax values of 440 ± 35 and 1,482 ± 110 fmol/mg of protein, respectively). The binding was temperature - and protein-dependent. Displacement studies showed a similar rank order of potency of various unlabeled ligands against both [3 H]RO 5-4864 and [3 H]PK 11195 (PK 11195 > Ro 5-4864 > flunitrazepam > flumazenil). These results suggest that peripheral-type benzodiazepine receptors are present in the retina of the rabbit, but not of the turtle. 相似文献
107.
K. Ogawa-Goto Y. Ohta K. Kubota N. Funamoto T. Abe T. Taki K. Nagashima† 《Journal of neurochemistry》1993,61(4):1398-1403
Abstract: Compositions of neutral and sulfated glucuronyl glycosphingolipids purified from human motor and sensory nerves and myelins were studied. Higher neutral glycosphingolipids (fraction B), which were separated from GazlCer (fraction A), were analyzed by TLC and TLC-immunostaining. Both nerve myelins contained paragloboside (nLc4 Cer) and nLc6 Cer dominantly as major higher glycosphingolipids and very little globoside (Gb4 Cer), whereas both nerves contained Gb4 Cer and nLc4 Cer. Besides these major glycosphingolipids, a neutral glycolipid containing asialoGMI (Gg4 Cer) epitope and other minor components such as ceramide trihexoside and ceramide dihexoside were detected in both nerves and their myelins. Furthermore, sulfated glucuronyl nLc4 Cer and n Lc6 Cer, which were monoclonal antibody HNK-1 reactive glycolipids, were detected in both nerves and myelins. 相似文献
108.
Laminin A, B1, and B2 Chain Gene Expression in Transected and Regenerating Nerves: Regulation by Axonal Signals 总被引:3,自引:0,他引:3
Manabu Doyu Gen Sobue Eishuku Ken Koji Kimata† Tamayuki Shinomura† Yoshihiko Yamada‡ Terunori Mitsuma Akira Takahashi 《Journal of neurochemistry》1993,60(2):543-551
Abstract: Laminin A, B1, and B2 chain mRNA levels in degenerating and regenerating mouse sciatic nerves were examined using northern blot analysis. In normal intact nerves, B1 and B2 mRNA steady-state levels were high, but when the nerves were crushed, the steady-state levels of B1 and B2 mRNA per milligram wet tissue weight of the distal segments of the nerves increased five- to eightfold over that of control levels as the total RNA and β-actin mRNA levels increased, suggesting that these increases were the consequence of Schwann cell proliferation after axotomy. When the steady-state levels of B1 and B2 mRNA were normalized as the ratio to total RNA or β-actin mRNA levels, however, they drastically decreased to about 20% of the normal nerve levels in the nerve segments distal to both the crush and transaction sites 1 day after injury. In the crushed nerves, B1 and B2 mRNA levels gradually increased as the regenerating nerves arrived at the distal segments and reestablished normal axon–Schwann cell contact, and then returned to normal levels on the 21 st day. In the transected nerves, where Schwann cells continued to be disconnected from axons, both B1 and B2 mRNA levels remained low. Cultured Schwann cells expressed detectable levels of B1 and B2 chain mRNA which significantly increased when the cells were cocultured with sensory neurons. However, mRNA for A chain was not detectable in the normal, axotomized nerves or in cultured Schwann cells. These data indicate that Schwann cells express laminin B1 and B2 chain mRNA that are up-regulated by axonal or neuronal contact, but they do not express A chain mRNA. 相似文献
109.
Twelve of 24 monospecific caprine reagents produced by absorption of alloimmune antisera identified a complex blood group
system of goats which was designated B, based on the results of a small comparison test with ovine reagents. The frequencies
of the 12 B factors differed significantly among the Australian Angora, Texan Angora, Cashmere, and Dairy goat breeds. Three
of the antigens detected by the reagents were shown to be related as linear subtypes, designated Ba1, Ba2, and Ba3, and inherited as alleles. The segregations of B factors in 80 sire groups involving 1086 offspring demonstrated that groups
of B factors (phenogroups) segregated as products of allelic genes.
This work was supported by a grant from the Australian Stud Book, Alison Road, Randwick, New South Wales 2031, Australia. 相似文献
110.
Tokuo Ogawa Junichi Sugenoya Norikazu Ohnishi Keiko Natsume Kazuno Imai Yoshikazu Kandori Akira Ishizuka Atsushi Osada 《European journal of applied physiology and occupational physiology》1993,67(4):354-359
We have examined the nonparallel changes in tampanic membrane temperatures (T
ty) from the two ears in response to various changes in body and head positions. Upon assuming a lateral recumbent position, the T
ty on the lower side increased while that on the upper side decreased. Pressure application over a wide area of the lateral chest only caused inconsistent and obscure asymmetric changes in T
ty. A lateral flexion of the head with the subject sitting upright and a rotation of the head to the side in a supine position induced an increase in the T
ty on the lower side compared to that on the upper side. The temperature and blood flow of the forehead often decreased on the lower side and increased on the upper side, although such responses were not always concomitant with the asymmetric changes in T
ty. A dorsal flexion of the head with the subject in a reclining position caused a slight increase in the T
ty, whereas raising the head upright induced a slight decrease in them. Two additional experiments were carried out with single photon emission computed tomography using 99mTc-hexamethylpropyleneamine oxime as tracer, and a slight, relative decrease in counts was noted in the right hemisphere during rotation of the head to the right. These results would strongly suggest that unilateral increases and decreases in T
ty could have been caused by one-sided decreases and increases, respectively, in blood flow to the brain and/or the tympanic membrane, induced by a vasomotor reflex involving vestibular stimulation. 相似文献