Search PubMed⌕ Search

Biomedical subjects

R M Gould

Publications and source records attributed to R M Gould.

At least 37 records · Page 2Linked to original sources
PubMed1993

A proteolipid protein gene family: expression in sharks and rays and possible evolution from an ancestral gene encoding a pore-forming polypeptide.

The myelin proteolipid proteins (PLP and DM20) are believed to act as "adhesive struts" in the extracellular apposition of the CNS myelin sheath. These proteins have been considered late evolutionary developments, which arose de novo in the antecedents of early tetrapods. However, PCR primed with degenerate oligonucleotides corresponding to common segments of rat PLP/DM20 revealed three novel mRNAs in the brains of two elasmobranchs. These mRNAs are closely related to each other and to mammalian DM20, but lack the sequence that distinguishes PLP from DM20. We term the novel proteolipid proteins DM alpha, DM beta, and DM gamma. At least DM alpha and DM gamma are highly expressed in white matter in myelinating shark brain. The DMs not only are highly homologous to each other, but also contain regions bearing similarities with segments of channel-forming regions of the nicotinic acetylcholine receptor and the glutamate receptor macromolecular complexes. Significantly, we find that across these segments, DM alpha and DM gamma are more similar to the channel proteins than the two channel proteins are to each other.

Amino Acid Sequence↗
PubMed1993

Basis for phospholipid incorporation into peripheral nerve myelin.

To characterize the mechanism(s) for targeting of phospholipids to peripheral nerve myelin, we examined the kinetics of incorporation of tritiated choline-, glycerol-, and ethanolamine-labeled phospholipids into four subfractions: microsomes, mitochondria, myelin-like material, and purified myelin at 1, 6, and 24 h after precursors were injected into sciatic nerves of 23-24-day-old rats. As validation of the fractionation scheme, a lag (> 1 h) in the accumulation of labeled phospholipids in the myelin-containing subfractions was found. This lag signifies the time between synthesis on organelles in Schwann cell cytoplasm and transport to myelin. In the present study, we find that sphingomyelin (choline-labeled) accumulated in myelin-rich subfractions only at 6 and 24 h, whereas phosphatidylserine (glycerol-labeled) and plasmalogen (ethanolamine-labeled) accumulated in the myelin-rich fractions by 1 h. The later phospholipids accumulate preferentially in the myelin-like fraction. These results are consistent with the notion that the targeting of sphingomyelin, a lipid present in the outer myelin leaflet, is different from the targeting of phosphatidylserine and ethanolamine plasmalogen, lipids in the inner leaflet. These findings are discussed in light of the possibility that sphingomyelin targeting is Golgi apparatus based, whereas phosphatidylserine and ethanolamine plasmalogen use a more direct transport system. Furthermore, the routes of phospholipid targeting mimic routes taken by myelin proteins P0 (Golgi) and myelin basic proteins (more direct).

Animals↗
PubMed1992

In situ hybridization with digoxigenin-labeled probes: sensitive and reliable detection method applied to myelinating rat brain.

A method for in situ hybridization of digoxigenin-labeled cDNA and cRNA probes to myelin protein mRNA is described. This technique has dual advantages of high structural resolution and high sensitivity and avoids problems associated with handling of radioactive materials. Furthermore, it can be readily combined in double labeling with immunocytochemical protein detection. We have used this technique to detect and locate mRNA for myelin basic protein (MBP), proteolipid protein (PLP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and myelin-associated glycoprotein (MAG) in oligodendrocytes of 7-day-old and adult rat brains. PLP and MAG mRNA were restricted to the perinuclear cytoplasm, whereas MBP and CNPase mRNA was additionally present in peripheral oligodendrocyte processes.

Animals↗
PubMed1992

Characterization of phospholipase A2 and acyltransferase activities in squid (Loligo pealei) axoplasm: comparison with enzyme activities in other neural tissues, axolemma and axoplasmic subfractions.

Phospholipase A2 and acyltransferase were assayed and characterized in pure axoplasm and neural tissues of squid. Intracellular phospholipase A2 activity was highest in giant fiber lobe and axoplasm, followed by homogenates from retinal fibers, optic lobe and fin nerve. In most preparations, exogenous calcium (5 mM) caused a slight stimulation of activity. EGTA (2 mM) was somewhat inhibitory, indicating that low levels of endogenous calcium may be required for optimum activity. Phospholipase A2 was inhibited by 0.1 mM p-bromophenacylbromide, and was completely inactivated following heating. The level of acylCoA: lysophosphatidylcholine acyltransferase activity was higher in axoplasm and giant fiber lobe than in other neural tissues of the squid. Km (apparent) and Vmax (apparent) for oleoyl-CoA and lysophosphatidylcholine were quite similar for axoplasm and giant fiber lobe enzyme preparations. Acyltransferase activity was inactivated by heat treatment, and greatly inhibited by 0.2 mM p-chloromercuribenzoate, and to a lesser extent by 20 mM N-ethylmaleimide. Phospholipase A2 activity was present in fractions enriched in axolemmal membranes (separated from squid retinal fibers and garfish olfactory nerve) from both tissues, and it was also highly concentrated in vesicles derived from squid axoplasm. In all three preparations, phospholipase A2 activity was stimulated by Ca++ (5 mM) and inhibited by EGTA (2 mM). In addition, axoplasmic cytosol (114,000 g supernatant) retained a substantial portion of a Ca(++)-independent phospholipase A2, active in the presence of 2 mM EGTA. Acyltransferase activity was present at high content in both axolemma membrane rich fractions, and among subaxoplasmic fractions and axoplasmic vesicles.

1-Acylglycerophosphocholine O-Acyltransferase↗
PubMed1991

Catalysis by yeast alcohol dehydrogenase.

Table 7 presents a brief summary of the effects of various mutations on some of the relevant kinetic constants. The results illustrate several important features of the use of site-directed mutagenesis in exploring structure and function of enzymes. Note that most of the mutations affect a given step or kinetic parameter in the mechanism, such as the binding of NAD+ or the turnover number with ethanol. Furthermore, one mutation can affect many steps in the mechanism. Thus, it is difficult to ascribe a particular role to an amino acid residue. It is also difficult to quantify the function of a residue, since the magnitudes of the effects on kinetic parameters will be modulated by the other amino acid residues that participate in the reaction. Comprehensive and quantitative kinetic studies of many mutant enzymes are required if we are to understand catalysis and specificity. We are reluctant to describe any residue as "essential" for activity since substitution with some amino acid can probably produce an enzyme with some residual activity. (Maybe the Thr48Gly enzyme would be active, as a water molecule could substitute for the hydroxyl of the threonine.) Likewise, when substitution of a residue partially, but not totally, decreases activity, it does not necessarily mean that the residue is "not essential". The change in activity can reflect the contribution of that residue to catalysis. On the other hand, if various substitutions of a residue do not change activity, it would be reasonable to conclude that the residue is not essential (Plapp et al., 1971). Most of the amino acid residues at the active site are involved in the catalytic mechanism, either by contacting the substrates directly or by participating in the chemistry. Some of the residues that are outside of the active site are indirectly involved, by affecting the structure of the protein. Substitution of an important amino acid residue should significantly affect activity, and studies on the kinetics and structure should allow one to distinguish among the various explanations.

Alcohol Dehydrogenase↗
PubMed1990

Substitution of arginine for histidine-47 in the coenzyme binding site of yeast alcohol dehydrogenase I.

Molecular modeling of alcohol dehydrogenase suggests that His-47 in the yeast enzyme (His-44 in the protein sequence, corresponding to Arg-47 in the horse liver enzyme) binds the pyrophosphate of the NAD coenzyme. His-47 in the Saccharomyces cerevisiae isoenzyme I was substituted with an arginine by a directed mutation. Steady-state kinetic results at pH 7.3 and 30 degrees C of the mutant and wild-type enzymes were consistent with an ordered Bi-Bi mechanism. The substitution decreased dissociation constants by 4-fold for NAD+ and 2-fold for NADH while turnover numbers were decreased by 4-fold for ethanol oxidation and 6-fold for acetaldehyde reduction. The magnitudes of these effects are smaller than those found for the same mutation in the human liver beta enzyme, suggesting that other amino acid residues in the active site modulate the effects of the substitution. The pH dependencies of dissociation constants and other kinetic constants were similar in the two yeast enzymes. Thus, it appears that His-47 is not solely responsible for a pK value near 7 that controls activity and coenzyme binding rates in the wild-type enzyme. The small substrate deuterium isotope effect above pH 7 and the single exponential phase of NADH production during the transient oxidation of ethanol by the Arg-47 enzyme suggest that the mutation makes an isomerization of the enzyme-NAD+ complex limiting for turnover with ethanol.

Alcohol Dehydrogenase↗
PubMed1990

Regional localization of RNA and protein metabolism in Schwann cells in vivo.

Schwann cells, which form and maintain extensive myelin sheaths, have the bulk of their lipid and protein synthesis restricted to the compact 'perinuclear' zone at the centre of the internode. Using teased fibre and quantitative electron microscopical autoradiography, we demonstrated that additional protein synthesis takes place in the lengthy processes of Schwann cell cytoplasm. This 'so-called' superficial cytoplasmic channel network forms a branching and anastomozing array that stretches between the perinuclear region and the distant paranodes. Protein synthesis apparently does not extend from this surface network into the Schmidt-Lanterman incisures or paranodal loops that circumscribe compact myelin. To maintain protein synthesis in these lengthy processes, Schwann cells transport a portion of their RNA along the superficial cytoplasmic channels at a rate (0.1 mm per day) that appears to be slightly lower than the transport rate reported for RNA along dendrites of hippocampal neurons in culture (0.5 mm per day). Nearly a week is required for labelled RNA to be transported from the Schwann cell nucleus to the paranodal terminals of the longer channels. The existence of this extended protein synthesis is not limited to myelinating Schwann cells. Schwann cell processes associated with small calibre axons also appear to synthesize some of their own proteins as the RNA needed to catalyze local translational events is transported into these processes.

Animals↗
PubMed1989

Use of lead aspartate block staining in quantitative EM autoradiography of phospholipids: application to myelinating peripheral nerve.

For quantitation of electron microscope (EM) autoradiographs, micrographs must contain clear images which are relatively free of heavy metal precipitates. Satisfactory contrast is usually obtained by staining individual ultra-thin sections with lead citrate. It was recently reported that sequential block staining of tissue with ferrocyanide-reduced osmium tetroxide and lead aspartate produced excellent contrast for EM autoradiography, with sections relatively free of lead precipitate. This protocol avoids the manipulation involved in staining individual ultra-thin sections. We have adapted this method to quantitative EM autoradiographic studies, primarily of phospholipid metabolism in peripheral nerve. We show that block staining with lead aspartate provides: (a) ultrastructural contrast of routinely high quality for myelinated peripheral nerve; (b) high (greater than 98%) retention of glycero-labeled lipid during dehydration and embedment; and (c) a distribution of de novo tritiated glycerol-labeled lipid in ultra-thin sections that is quantitatively identical to the distribution recorded for samples stained by the more laborious post-embedment method. During a 2-hr labeling period in vivo, tritiated glycerol is incorporated into phosphatidylcholine (44%), phosphatidylethanolamine (22%), other phospholipids (16%), and neutral lipids (15%). The analysis of grain distribution in developing sciatic nerve labeled for 2 hr with tritiated glycerol demonstrates that myelinating Schwann cells play the major role in synthesis of endoneurial lipids. Lipid synthesis in myelinated fibers is localized in perinuclear regions of Schwann cell cytoplasm. These regions lie external to compact myelin. Unmyelinated fibers and other endoneurial cells independently incorporate glycerol into lipids.

Animals↗
PubMed1988

Documentation of time expenditures of clinical dietitians: results of a statewide time study in Texas.

The purpose of this study was to establish a profile of the amount of time expended by clinical dietitians in providing nutrition services. One hundred twenty-seven dietitians in 49 hospitals in Texas collected time data according to nutrition care activity, diet order classification, care level, and diagnosis. Frequency, mean time, standard deviation, and percentage of time were computed for each of the variables. Data were analyzed to ascertain relationships among the variables. The dietitians expended 50.7% of their time performing client-related activities, 9.8% in administrative/managerial functions, 1.0% in professional activities, 5.1% in non-professional activities, 14.0% in delay activities, and 19.5% in transit time. The diet order classification that required the largest amount of the dietitians' time according to selected components of the nutrition care process was the calorie-controlled diabetic diet. Among the four nutrition care levels, dietitians expended the greatest percentages of time providing services for patients requiring intermediate (35.4%) and advanced intermediate care (30.4%). Dietitians expended more time providing client-related activities to patients diagnosed with diseases and disorders of the endocrine and excretory systems than to other patients. Mean times provide the clinical manager with documentation for determining staffing needs and costs of services.

Dietetics↗
PubMed1988

Deposition and transfer of axonally transported phospholipids in rat sciatic nerve.

Radioactive glycerol, ethanolamine, or choline injected into the vicinity of the cell bodies of rat sciatic nerve sensory fibers is incorporated into phospholipid. Some newly synthesized ethanolamine and choline phosphoglycerides are subsequently committed to transport down the sciatic nerve axons at a rate of several hundred millimeters per day. Most labeled choline phosphoglycerides move uniformly down the axons; in contrast, the crest of moving ethanolamine phosphoglycerides is continually attenuated. These data, as well as differences in the clearance of these phospholipids distal to a nerve ligature, suggest that various classes of labeled phospholipids are differentially unloaded from the transport vector (possibly by exchange with unlabeled lipid in stationary axonal structures) during movement down the axons. The extent of unloading appears to be defined by the base moiety; both diacyl and plasmalogen species of ethanolamine phosphoglycerides exchange extensively with stationary axonal lipids, while most choline phosphoglycerides continue down the axons. Autoradiographic studies with 3H-choline and 3H-ethanolamine demonstrated that most unloaded phospholipid is initially deposited in axonal structures; some of this unloaded lipid is subsequently transferred to the axon/myelin interface (axolemma?) and then to myelin. Although transported ethanolamine phosphoglycerides exchange more extensively with lipids in stationary axonal structures than do choline phosphoglycerides, at early times more label from 3H-choline is found in myelin. A model to resolve this seeming discrepancy is proposed, wherein a differential topographic localization of phospholipid classes in the membrane of the transport vector allows for a preferential extensive exchange of transported ethanolamine phosphoglycerides with lipids in stationary axonal structures, while choline phosphoglycerides become available for rapid transfer to myelin by a process involving vesicle fusion with axolemma.

Animals↗
PubMed1987

Lipid metabolism in various regions of squid giant nerve fiber.

The purpose of this investigation was to compare the incorporation of radioactivity from various precursors into lipids of different regions of squid giant nerve fiber systems including axoplasm, axon sheath, giant fiber lobes which contain stellate ganglion cell bodies, and the remaining ganglion including giant synapses. To identify the labeled lipids, stellate ganglia including giant fiber lobes and the remaining tissue were first incubated separately with [14C]glucose, [32P]phosphate, [14C]serine, [14C]acetate and [3H]myristate. The radioactivity from glucose, after conversion to glycerol and fatty acids, was incorporated into most lipids, including triacylglycerol, free fatty acids, cardiolipin, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylserine, sphingomyelin and ceramide 2-aminoethylphosphanate [corrected]. The radioactivity from serine was largely incorporated into phosphatidylserine and, to a lesser extent, into other phospholipids, mainly as the base component. The sphingoid bases of ceramide and sphingomyelin were also significantly labeled. Saturated and monounsaturated and, to a lesser extent, polyunsaturated fatty acids of these lipids were synthesized from acetate, glucose and myristate. Among the major lipids, cholesterol was not labeled by any of the radioactive compounds used. Ganglion residues incorporated the most radioactivity in total lipids from either [14C]glucose or [14C]serine, followed by giant fiber lobes and then sheath. Axoplasm incorporated the least. Among various lipids, phosphatidylethanolamine with shorter saturated fatty acids and phosphatidylglycerol contained the most radioactivity from glucose in all regions. Axoplasm was characterized by a higher proportion of glucose radioactivity in ceramide, sphingomyelin and phosphatidylglycerol. Axoplasm and sheath contained a higher proportion of serine radioactivity than did the other two regions in ceramide. Essentially no radioactivity from [14C]galactose was incorporated in any region.

Acetates↗
PubMed1987

Kinetic characterization of yeast alcohol dehydrogenases. Amino acid residue 294 and substrate specificity.

A three-dimensional model of yeast alcohol dehydrogenase, based on the homologous horse liver enzyme, was used to compare the substrate binding pockets of the three isozymes (I, II, and III) from Saccharomyces cerevisiae and the enzyme from Schizosaccharomyces pombe. Isozyme I and the S. pombe enzyme have methionine at position 294 (numbered as in the liver enzyme, corresponding to 270 in yeast), whereas isozymes II and III have leucine. Otherwise the active sites of the S. cerevisiae enzymes are the same. All four wild-type enzymes were produced from the cloned genes. In addition, oligonucleotide-directed mutagenesis was used to change Met-294 in alcohol dehydrogenase I to leucine. The mechanisms for all five enzymes were predominantly ordered with ethanol (but partially random with butanol) at pH 7.3 and 30 degrees C. The wild-type alcohol dehydrogenases and the leucine mutant had similar kinetic constants, except that isozyme II had 10-20-fold smaller Michaelis and inhibition constants for ethanol. Thus, residue 294 is not responsible for this difference. Apparently, substitutions outside of the substrate binding pocket indirectly affect the interactions of the alcohol dehydrogenases with ethanol. Nevertheless, the substitution of methionine with leucine in the substrate binding site of alcohol dehydrogenase I produced a 7-10-fold increase in reactivity (V/Km) with butanol, pentanol, and hexanol. The higher activity is due to tighter binding of the longer chain alcohols and to more rapid hydrogen transfer.

Alcohol Oxidoreductases↗
PubMed1987

Specificity of the attenuation response of the threonine operon of Escherichia coli is determined by the threonine and isoleucine codons in the leader transcript.

Expression of the threonine (thr) operon enzymes of Escherichia coli is regulated by an attenuation mechanism. The regulatory portion of the operon contains a region coding for a leader peptide that contains consecutive threonine and isoleucine codons. It is thought that translation of the leader peptide controls the frequency of transcription termination at the attenuator site. Using oligonucleotide-directed site-specific mutagenesis we have altered the putative control codons of the leader peptide coding region. In two of the mutants the threonine and isoleucine codons were changed to produce peptides containing histidine and tyrosine codons. Both mutants showed loss of regulation by threonine and isoleucine. A hisT mutation, which leads to an undermodification of tRNA(His), increased thr operon expression in the mutants threefold but did not affect expression of the wild-type thr operon. Two other mutants were constructed that contained two histidine codons early in the leader peptide. Expression in both of these mutants was unaltered by the presence of the hisT allele or by the addition of threonine and isoleucine to the growth medium. In addition, a wild-type strain containing a temperature-sensitive threonyl-tRNA synthetase mutation showed increased thr operon expression at the non-permissive temperature, whereas none of the mutants showed any change. Taken together these data indicate that the specificity of the attenuation response is effected by specific control codons within the thr leader peptide coding region. We have also directly demonstrated thr leader peptide synthesis in vitro using a plasmid encoding the wild-type thr leader region to direct the synthesis of a peptide of the appropriate molecular weight when labeled with [3H]threonine but not with [3H]histidine or [3H]tyrosine. Conversely, when extracts were incubated with templates containing the mutated DNAs, peptides were labeled that showed patterns consistent with the expected amino acid compositions. These data indicate that the thr leader RNA is translated into the predicted leader peptide.

Codon↗
PubMed1987

Phospholipid metabolism in mouse sciatic nerve in vivo.

To probe the activities of various pathways of lipid metabolism in peripheral nerve, six phospholipid-directed precursors were individually injected into the exposed sciatic nerves of adult mice, and their incorporation into phospholipids and proteins was studied over a 2-week period. Tritiated choline, inositol, ethanolamine, serine, and glycerol were mainly used in phospholipid synthesis; in contrast, methyl-labeled methionine was primarily incorporated into protein. Phosphatidylcholine was the main lipid formed from tritiated choline, glycerol, and methionine precursors. Phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol were the main lipids formed from serine, ethanolamine, and inositol, respectively. With time there was a shift in label among phospholipids, with higher proportions of choline appearing in sphingomyelin, glycerol in phosphatidylserine, ethanolamine in phosphatidylethanolamine (plasmalogen), and inositol in polyphosphoinositides, especially phosphatidylinositol 4,5-bisphosphate. We suggest that the delay in formation of these phospholipids, which are concentrated in peripheral nerve myelin, may, at least in part, be due to their formation at a site(s) distant from the sites where the bulk of Schwann cell lipids are made. We propose that separating the synthesis of these myelin-destined lipids to near the Schwann cell's plasma membrane would facilitate their concentration in peripheral nerve myelin sheaths. At earlier labeling times, ethanolamine and glycerol were more actively incorporated into phosphatidylcholine and phosphatidylinositol, respectively, than later. The transient labeling of these phospholipids may reflect some unique role in peripheral nerve function.

Animals↗
PubMed1987

Localization of phospholipid synthesis to Schwann cells and axons.

Quantitative electron microscopic autoradiography was used to detect and characterize endoneurial sites of lipid synthesis in mouse sciatic nerve. Six tritiated phospholipid precursors (choline, serine, methionine, inositol, glycerol, and ethanolamine) and a protein precursor (proline) were individually injected into exposed nerves and after 2 h the mice were perfused with buffered aldehyde. The labeled segments of nerve were prepared for autoradiography with procedures that selectively remove nonincorporated precursors and other aqueous metabolites, while preserving nerve lipids (and proteins). At both the light and electron microscope levels, the major site of phospholipid and protein synthesis was the crescent-shaped perinuclear cytoplasm of myelinating Schwann cells. Other internodal Schwann cell cytoplasm, including that in surface channels, Schmidt-Lanterman incisures, and paranodal regions, was less well labeled than the perinuclear region. Newly formed proteins were selectively located in the Schwann cell nucleus. Lipid and protein formation was also detected in unmyelinated fiber bundles and in endoneurial and perineurial cells. Tritiated inositol was selectively incorporated into phospholipids in both myelinated axons and unmyelinated fibers. Like inositol, glycerol incorporation appeared particularly active in unmyelinated fibers. Quantitative autoradiographic analyses substantiated the following points: myelinating Schwann cells dominate phospholipid and protein synthesis, myelinated axons selectively incorporate tritiated inositol, phospholipid precursors label myelin sheaths and myelinated axons better than proline.

Animals↗
PubMed1987

Structure and function in yeast alcohol dehydrogenases.

The structure and kinetics of the isozymes from Saccharomyces cerevisiae (ADH I, II, III) have been compared, and the ADH I gene was specifically mutagenized in order to substitute amino acid residues of particular interest. A model of the yeast enzyme was constructed on the basis of the structure of the homologous horse liver enzyme. Steady state kinetic studies, at pH 7.3 and 30 degrees C, showed that the enzymes follow the Ordered Bi Bi mechanism. ADH II has a Michaelis constant for ethanol that is 10-fold smaller than the constants found for ADH I or III. Replacement of Met-294 (liver numbering) in the substrate binding pocket of ADH I with Leu, as found in ADH II, could be responsible for the different kinetics. However, the mutant enzyme, ADH I-Leu, had constants with ethanol that were similar to those of ADH I. Nevertheless, the Leu enzyme had better catalytic activity with longer chain alcohols than did the Met enzyme. Other substitutions must account for the differences between ADH I and II. His-47 binds the pyrophosphate of coenzyme, and replacement with Arg (as in the liver enzyme) decreases turnover numbers by 6-fold and dissociation constants for NAD+ and NADH by only 2 to 4-fold. The lower turnover number explains why yeast harboring the mutant Arg enzyme are resistant to poisoning by allyl alcohol. ADH I and ADH I-Arg have maximal activity on ethanol at pH values above a pK of about 7. Replacement of His-51 with Gln reduces activity 12-fold and abolishes the pK value.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcohol Dehydrogenase↗
PubMed1985

Prostanoid synthesis in peripheral nerve.

The transformation of [1-14C]arachidonic acid into radiolabeled prostanoids was studied with homogenates and desheathed sciatic nerves of rats and frogs. All of the preparations studied were shown to synthesize prostaglandins; the specific prostanoids made were characterized by their migration on thin-layer chromatograms in three separate solvent systems. Both desheathed rat nerve and homogenates synthesize prostaglandin E2, prostaglandin F2 alpha, prostaglandin D2, 6-ketoprostaglandin F1 alpha and thromboxane B2. With preparations from frog nerve, prostaglandin E2 was the major prostanoid product formed. Several conditions were able to modulate the production of prostaglandin E2 with desheathed frog nerve. Electrical stimulation at high frequency (100 Hz) for 30 min increased the formation of labeled prostaglandin E2. Inclusion of glutathione also affected prostaglandin E2 formation. A lower concentration (0.1 mM) stimulated prostaglandin synthesis, while 1 mM glutathione was partially inhibitory. In both the rat and frog system, prostanoid synthesis was suppressed by indomethacin and aspirin.

Animals↗