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The Journal of General Physiology Oct 1976The effects of pulse application of puromycin (PURO) or cycloheximide (CHX) were tested on the circadian rhythm (CR) of spontaneous compound action potential (CAP)...
The effects of pulse application of puromycin (PURO) or cycloheximide (CHX) were tested on the circadian rhythm (CR) of spontaneous compound action potential (CAP) activity in the isolated Aplysia eye. CAP activity was recorded from the optic nerve in constant darkness at 15degreesC. PURO pulses (6, 12 h; 12--134 mug/ml) and CHX pulses (12 h, 500--2,000 mug/ml) caused dose-dependent phase delays in the CR when administered during projected night. PURO pulses (6 h, 125 mug/ml) caused phase advances when given during projected day and caused phase delays when given during projected night. In biochemical experiments PURO (12 h, 20 mug/ml) and CHX (12 h, 500 mug/ml) inhibited leucine incorporation into the eye by about 50%. PURO (12 h; 50, 125 mug/ml) also changed the molecular weight distribution of proteins synthesized by the eye during the pulse. The effect of PURO (12 h, 125 mug/ml) on the level of incorporation was almost completely reversible within the next 12 h but the phase-shifted eye showed an latered spectrum of proteins for up to 28 h after the pulse. In electrophysiological experiments spontaneous CAP activity and responses to light were measured before, during, and after drug treatments. In all, eight parameters in three periods were analyzed quantitatively. Of these 24 indices, only 3 showed significant changes. PURO increased spontaneous CAP frequency by 67% 0-7 h after the drug pulse and increased the CAP amplitude of the tonic light response by 23% greater than 7 h after the pulse. CHX increased the intraburst spontaneous CAP frequency by 33% during the pulse and CAP frequency of the tonic light response by 32% 0-7 h after the pulse. The above data indicate that phase-shifting doses of PURO and CHX inhibit protein synthesis in the eye without causing adverse electrophysiological effects, and suggest that protein synthesis is involved in the production of the CR of the isolated Aplysia eye.
Topics: Action Potentials; Animals; Circadian Rhythm; Cycloheximide; Depression, Chemical; Dose-Response Relationship, Drug; Electrophysiology; Kinetics; Leucine; Mollusca; Nerve Tissue Proteins; Neurons; Optic Nerve; Puromycin
PubMed: 993764
DOI: 10.1085/jgp.68.4.359 -
Biochemical and Biophysical Research... Jun 1965
Topics: Arginine; Canavanine; Culture Techniques; DNA, Viral; HeLa Cells; Histones; Nucleotides; Poliovirus; Puromycin; Thymidine
PubMed: 4284637
DOI: 10.1016/0006-291x(65)90321-9 -
The Journal of Biological Chemistry Nov 1978A new, rapid, and sensitive assay for phospholipase A, utilizing commercially available [14C]phosphatidylethanolamine with 14C label in both palmitic acid moieties, was...
A new, rapid, and sensitive assay for phospholipase A, utilizing commercially available [14C]phosphatidylethanolamine with 14C label in both palmitic acid moieties, was used to study phospholipase A release from perfused liver, hepatocytes, and intestinal cells from rats. Heparin triggered a prompt release of phospholipase A from perfused liver. Phospholipase A and triglyceride lipase were released from hepatocytes at a linear rate for 1 h and 30 min, respectively. Heparin (20 u/ml) doubled the release of phospholipase A and triglyceride lipase from hepatocytes. Colchicine (0.1 mM), but not puromycin (0.2 mM), inhibited basal and heparin-stimulated phospholipase A release by 40%. Since the amount of phospholipase A and triglyceride lipase released into the medium greatly exceeded intracellular activities, it is possible that secretion is coupled with intracellular conversion from inactive to active forms of the enzymes. Dibutyryl cyclic AMP (1 mM) inhibited phospholipase A (48%) and triglyceride lipase (82%) release from hepatocytes. Epinephrine, dexamethasone, and clofibrate inhibited release of triglyceride lipase but not phospholipase A. Phospholipase A activity of intestinal cells was greater than in hepatocytes, but neither heparin nor dibutyryl cyclic AMP affected phospholipase A release from intestinal cells. These results suggest that the liver is a major source of phospholipase A of postheparin plasma. The fact that dibutyryl cyclic AMP affects the release of these enzymes suggests an additional mechanism for hormonal regulation of lipid and lipoprotein metabolism.
Topics: Animals; Colchicine; Dinitrophenols; Heparin; Hypoxia; Kinetics; Lipase; Liver; Male; Perfusion; Phospholipases; Puromycin; Rats
PubMed: 701283
DOI: No ID Found -
Fertility and Sterility Jul 2001To determine the effect of sequential oocyte activation with calcium ionophore A23187 (A23187) and puromycin on oocytes that were unfertilized after intracytoplasmic...
OBJECTIVE
To determine the effect of sequential oocyte activation with calcium ionophore A23187 (A23187) and puromycin on oocytes that were unfertilized after intracytoplasmic sperm injection (ICSI).
DESIGN
Laboratory examination.
SETTING
The in vitro fertilization/embryo transfer unit in Tokushima University Hospital.
PATIENT(S)
Discarded unfertilized oocytes following ICSI.
INTERVENTION(S)
All 172 oocytes that showed no evidence of normal fertilization 18 hours after ICSI were exposed to A23187 (5 microM) for 5 minutes and consequently were treated with puromycin (10 microg/mL) for 5 hours.
MAIN OUTCOME MEASURE(S)
The activation rate, proportion of oocytes that showed two pronuclei with the second polar body (2PN2PB), and cleavage rate were calculated. Chromosomal analysis of the oocytes with 2PN2PB was also performed.
RESULT(S)
The treatment activated 146 out of 172 oocytes (84.9%) and 30.1% of the activated oocytes showed 2PN2PB. Sixteen of 25 oocytes with 2PN2PB (64.0%) cleaved. There was no difference in the activation rate, proportion of activated oocytes with 2PN2PB, or cleavage rate between oocytes that were injected with a motile spermatozoon and those injected with an immotile spermatozoon. Chromosomal analysis showed a normal diploid set of chromosomes (n = 46) in four of five analyzable oocytes.
CONCLUSION(S)
The sequential treatment of calcium ionophore A23187 and puromycin activates unfertilized oocytes after ICSI. The resultant oocytes with 2PN2PB can cleave.
Topics: Calcimycin; Cellular Senescence; Chromosome Mapping; Cleavage Stage, Ovum; Diploidy; Female; Fertilization in Vitro; Humans; Ionophores; Male; Oocytes; Puromycin; Sperm Injections, Intracytoplasmic; Sperm Motility; Spermatozoa; Treatment Failure
PubMed: 11438334
DOI: 10.1016/s0015-0282(01)01839-8 -
Antimicrobial Agents and Chemotherapy Oct 1975The effect of a series of puromycin analogues and aminoacyl chloramphenicol derivatives on poly(U,C)-directed polyphenylalanine synthesis in an Escherichia coli...
The effect of a series of puromycin analogues and aminoacyl chloramphenicol derivatives on poly(U,C)-directed polyphenylalanine synthesis in an Escherichia coli cell-free system was examined. A comparison between the structures and activities of the puromycin and chloramphenicol analogues was made to examine the proposal that ribosomal binding sites for both antibiotics overlap. Our results suggest that the dichloroacetamido group in the chloramphenicol molecule does not correspond to the role of the aminoacyl moieties of either puromycin or aminoacyl transfer ribonucleic acid. These results comparing the structures and activities of puromycin and chloramphenicol analogues also seem inconsistent with a common binding site for the p-substituted phenyl moieties of the two antibiotics. Previous data have indicated that both sites are mutually affected by the prior binding of either antibiotic. Although it is possible that chloramphenicol and puromycin may have overlapping bindings sites, no common structural features between the two antibiotics are supported by our data.
Topics: Binding Sites; Cell-Free System; Chloramphenicol; Escherichia coli; Peptide Chain Initiation, Translational; Phenylalanine; Puromycin; Structure-Activity Relationship
PubMed: 1103725
DOI: 10.1128/AAC.8.4.439 -
The Journal of Toxicological Sciences Feb 1979The suppressive effect of dipyridamole on the proteinuria of aminonucleoside nephrosis and protamine-induced proteinuria, was investigated. Male Wistar rats were given...
The suppressive effect of dipyridamole on the proteinuria of aminonucleoside nephrosis and protamine-induced proteinuria, was investigated. Male Wistar rats were given puromycin aminonucleoside (80 mg/kg s.c.) or protamine sulfate (20 mg/kg i.v.), and the urine was collected in metabolic cages. The content of proteins in the urine was determined by using a continuous gradient microgel electrophoresis procedure. Dipyridamole (20 mg/kg p.o.) suppressed the excretion of albumin and proteins larger than albumin (HMP) in aminonucleoside nephrosis. But the excertion of proteins smaller than albumin (LMP) was not affected by dipyridamole. Dipyridamole also suppressed the excertion of HMP in protamine-induced proteinuria, though the excretion of albumin and LMP was not affected. Puromycin aminonucleoside and protamine sulfate were known to cause renal glomerular epithelial changes referred to as "fusion" of foot processes. Since dipyridamole was effective in suppressing the both types of proteinuria, this drug was considered to improve the damaged renal glomerular barrier for plasma proteins.
Topics: Albuminuria; Animals; Dipyridamole; Male; Nephrosis; Protamines; Proteinuria; Puromycin; Puromycin Aminonucleoside; Rats; Time Factors
PubMed: 480401
DOI: 10.2131/jts.4.1 -
Infection and Immunity Jun 1982Dissemination of Leishmania within the host is related to parasites undergoing unchecked proliferation. We therefore studied the effects of oxidant generating systems on...
Dissemination of Leishmania within the host is related to parasites undergoing unchecked proliferation. We therefore studied the effects of oxidant generating systems on promastigote multiplication by (i) direct determinations of organism proliferation and (ii) the incorporation of [3H]uracil into promastigote nucleoprotein. These two parameters correlated closely as measures of organism replication as demonstrated by parallel suppression of them by the protein synthesis inhibitors puromycin and cycloheximide and the nucleic acid synthesis inhibitors actinomycin D and mitomycin C. Promastigotes showed dose-related susceptibility to reagent and generated hydrogen peroxide (H2O2) as reflected in quantitatively similar decreases in multiplication and [3H]uracil incorporation. These effects were specific for H2O2 as catalase abrogated the dimunition in multiplication. The generation of superoxide anion by acetaldehyde-xanthine oxidase (10 mU/ml) did not alter promastigote replication or nucleoprotein synthesis. These results indicate that Leishmania donovani promastigotes are damaged by H2O2 and that the incorporation of [3H]uracil into promastigote nucleoprotein may be useful for studying the interaction of this parasite with host effector cells.
Topics: Acetaldehyde; Animals; Cycloheximide; Dactinomycin; Leishmania; Oxidation-Reduction; Oxygen; Peroxides; Puromycin; Superoxides; Xanthine Oxidase
PubMed: 6284640
DOI: 10.1128/iai.36.3.1023-1027.1982 -
Fertility and Sterility Nov 2012To evaluate the dynamics of the nuclear maturation (NM) of in vitro-matured (IVM) oocytes and to determine the most favorable duration of meiosis II (MII) arrest in...
OBJECTIVE
To evaluate the dynamics of the nuclear maturation (NM) of in vitro-matured (IVM) oocytes and to determine the most favorable duration of meiosis II (MII) arrest in relation to the normal activation response.
DESIGN
Experimental.
SETTING
University-affiliated infertility clinic.
PATIENT(S)
Donated immature germinal vesicle oocytes (GV).
INTERVENTION(S)
The GV underwent spontaneous IVM and the dynamics of NM studied by real-time monitoring. The IVM oocytes were parthenogenetically activated at different MII arrest points and their response assessed.
MAIN OUTCOME MEASURE(S)
Moment of GV breakdown; extrusion of the first polar body; duration of MI and MII arrest; activation rate (AR) and type.
RESULT(S)
Two GV populations-early (E-IVM, 18.4 ± 2.7 hours) and late (L-IVM, 26.3 ± 3.8 hours) maturing-were defined according to the time required for extrusion of the first polar body. Significantly more E-IVM than L-IVM exhibited a normal activation response (61.3% vs. 34.6%), but AR were similar (average, 88.6%) in both groups. Duration of the GV stage differed between the two groups, but MI arrest (14.0 ± 0.3 hours) was constant. The E-IVM arrested at MII for at least 4.3 hours displayed significantly lower AR and similar normal activation rates (61.3%) to E-IVM arrested for a shorter time (83.9% vs. 100%). The L-IVM displayed a similar AR (80.8%), but lower normal activation rates than E-IVM (34.6%), regardless of when activation took place.
CONCLUSION(S)
The success of IVM depends on the NM timing rather than on the length of MII arrest.
Topics: Adult; Cells, Cultured; Female; Humans; In Vitro Oocyte Maturation Techniques; Meiosis; Oocyte Donation; Oocytes; Parthenogenesis; Polar Bodies; Puromycin; Time Factors; Time-Lapse Imaging; Young Adult
PubMed: 22901848
DOI: 10.1016/j.fertnstert.2012.07.1116 -
Pancreas Oct 2019
Topics: Acinar Cells; Animals; Antimetabolites, Antineoplastic; Autophagy; Humans; Male; Microscopy, Electron, Transmission; Pancreas; Pancreatitis; Puromycin; Rats, Wistar; Vacuoles
PubMed: 31609938
DOI: 10.1097/MPA.0000000000001389 -
The Biochemical Journal Sep 1967The properties of a site, on Escherichia coli ribosomes, which binds peptidyl-s-RNA (where s-RNA refers to ;soluble' or transfer RNA) have been investigated. The binding...
The properties of a site, on Escherichia coli ribosomes, which binds peptidyl-s-RNA (where s-RNA refers to ;soluble' or transfer RNA) have been investigated. The binding is stable both in low Mg(2+) concentration (0.1mm), and in high Mg(2+) concentration (10mm) in the absence or presence of potassium chloride (86mm). Puromycin has been used to break the bond between the s-RNA and the polypeptide, and in the absence of further protein synthesis this technique exposes free s-RNA molecules on the ribosomes. The s-RNA exposed remains bound in high Mg(2+) concentration, but the binding is unstable in high Mg(2+) concentration with potassium chloride and the s-RNA can be freed completely from the ribosomes by lowering the Mg(2+) concentration. It can also be displaced by s-RNA in the medium. It is suggested that this ribosomal binding site for peptidyl-s-RNA is the site for peptide bond formation. Further, it is proposed that it is the same site that can be demonstrated on ribosomes not engaged in protein synthesis and that, in high Mg(2+) concentration, will bind s-RNA molecules charged or uncharged with amino acids.
Topics: Binding Sites; Carbon Isotopes; Escherichia coli; Leucine; Magnesium; Peptides; Phosphates; Phosphorus Isotopes; Puromycin; RNA, Bacterial; RNA, Transfer; Ribosomes
PubMed: 4860641
DOI: 10.1042/bj1040934