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Biomedical subjects

G Y Sun

Publications and source records attributed to G Y Sun.

At least 37 records · Page 2Linked to original sources

Chronic ethanol and iron administration on iron content, neuronal nitric oxide synthase, and superoxide dismutase in rat cerebellum.

Excessive chronic ethanol administration to animals has been shown to cause oxidative insults to many body organs, including the liver and brain. In many instances, iron supplementation to the diet may further aggravate ethanol-induced liver damage. However, whether increased dietary iron can enhance the damage in the brain is unknown. In this study, four groups of Sprague-Dawley rats were fed a Lieber-DeCarli liquid diet containing 5% (w/v) ethanol or isocaloric amount of maltase and/or 0.25% (w/v) carbonyl iron for 2 months. At the end of the feeding regimen, iron contents were determined in the plasma, liver, cerebral cortex, and cerebellum. Cerebellar superoxide dismutase (SOD) and nitric oxide synthase (NOS) activities were measured and mRNA levels of MnSOD, CuZnSOD, and nNOS in the cerebellar granule cell layer were quantitated by in situ hybridization. Ethanol treatment alone caused an increase in iron levels in plasma, no change in the liver and cerebral cortex, but a decrease in the cerebellum. Iron supplementation increased liver iron >4-fold but did not alter iron contents in the cerebellum and cortex. All of the mRNA species examined and SOD activity were not affected by either iron or ethanol administration. However, NOS activity in the cerebellum was significantly enhanced by ethanol, whereas iron supplementation had an opposite effect. Our results indicate that iron supplementation to animals consuming ethanol may have tissue-specific effects. Furthermore, ethanol-induced increase in NOS activity in the cerebellum may explain the sensitivity of cerebellar neurons to oxidative insult.

Animals↗

STR HUMARA locus gene and genotype frequencies in Han and Bei populations in China.

For the purpose of the population genetics study of the HUMARA locus, the allele, and genotype frequencies were determined in two Chinese population samples (Han-101, Bei-113) using PCR, PAGE, and silver staining. Fourteen alleles were found. The size of amplified fragments were 258 bp-315 bp. The observed heterozygosities were 0.83 in the Han population and 0.73 in the Bei population respectively. The expected heterozygosities were 0.91 in the Han population and 0.97 in the Bei population respectively. Both populations meet Hardy-Weinberg expectation, Han population x2 = 17.7206, df = 11, p > 0.05; Bei population x2 = 7.4268, df = 10, p > 0.05. The discrimination power were 0.95 in females and 0.89 in males in the Han population, 0.94 in females and 0.88 in males in the Bei population. Thus, the allelic frequency data can be used in the personal identification and parentage testing in the forensic science practice. The PCR test established in this study is robust and reproducible.

Alleles↗

Age differences in the expression of metabotropic glutamate receptor 1 and inositol 1,4,5-trisphosphate receptor in mouse cerebellum.

Age differences in the expression of cerebellar metabotropic glutamate receptor 1 (mGluR1) and inositol 1,4,5-trisphosphate receptor (IP3R) were investigated using male C57BL/6NNIA mice 5, 15 and 24 months of age. In situ hybridization for mGluR1 mRNA in the granule cell layer indicated significantly higher mRNA levels in the 24-month-old group as compared to the 5- and 15-month-old groups. However, mRNA levels of individual Purkinje neurons did not show age differences. Western blot analysis using antibody against the predominant isoform, mGluR1a, showed a decline in protein levels in the 24-month-old animals. In situ hybridization for IP3R type 1 mRNA in Purkinje neurons showed a slight but not significant decline in the 24-month-old group. Further assay of [3H]IP3 binding with cerebellar membranes showed significant reduction in Bmax values in the 15- and 24-month-old groups as compared to the 5-month-old group but Kd values were not changed. The decrease in mGluR1a receptor protein together with reduction in IP3R binding sites may play an important role in the decline in cerebellar functions with increasing age.

Aging↗

Dietary supplementation of grape polyphenols and chronic ethanol administration on LDL oxidation and platelet function in rats.

Polyphenolic compounds have been implicated as the active ingredients for the cardiac protective effect in red wine. We tested the effects of dietary supplementation of polyphenols from grape (GP) and chronic ethanol administration on low-density-lipoprotein (LDL) oxidation and platelet function in rats. Four groups of young male Sprague-Dawley rats were fed the following diets for 2 months: (I) a high fat Lieber-DeCarli liquid diet with an isocaloric amount of maltose, (II) with 5% ethanol (w/v), (III) with 5 mg/dL of GP, and (IV) ethanol plus GP. Platelet aggregation was induced by thrombin and phorbol myristate acetate (PMA) and LDL oxidation was induced by Cu2+. Chronic ethanol administration resulted in a significant increase in LDL oxidation and this effect was partially protected by supplementation with GP. Although platelet number was not affected by either ethanol or GP administration, platelet aggregation induced by thrombin was reduced in ethanol, GP and ethanol plus GP groups as compared to controls. On the other hand, platelet aggregation induced by PMA was not altered in any groups, suggesting that protein kinase C was not a causal factor for the reduction of aggregatory response induced by thrombin. These results show similar effects of ethanol and GP on platelet aggregation but different effects on LDL oxidation. It can be concluded that dietary supplementation with GP may exert partial protection on oxidative insults such as those elicited by chronic ethanol ingestion.

Animals↗

Changes in IP3R1 and SERCA2b mRNA levels in the gerbil brain after chronic ethanol administration and transient cerebral ischemia-reperfusion.

Despite epidemiological studies indicating a positive relationship between alcohol and stroke, little is known with regard to effect of chronic alcohol on neuronal injury after stroke. In this study, we examined the effect of chronic ethanol on mRNA levels of sarcoplasmic or endoplasmic Ca2+-ATPase (SERCA2b) and inositol 1,4, 5-triphosphate receptor (IP3R1) in gerbils subjected to global cerebral ischemia induced by ligation of both common carotid arteries. Gerbils were given daily by intragastric intubation either a liquid diet containing ethanol (4 g/kg) or the same diet with an isocaloric amount of sucrose for 35 days. They were subsequently subjected to a 5 min ischemic insult followed by reperfusion for 48 h. In agreement with other studies, ischemic insult caused significant decreases (P<0.05) in mRNA levels of both IP3R1 and SERCA2b in the hippocampal CA1 region but not in the dentate gyrus. Nevertheless, despite a significant (P<0.05) decrease in SERCA2b mRNA in the Purkinje neurons, chronic ethanol did not alter the expression of this mRNA species in the hippocampal CA1 neurons nor did it alter the decrease in SERCA2b mRNA due to cerebral ischemic insult. Since IP3R1 and SERCA2b are key mediators for regulation of intracellular Ca2+ stores, the decrease in SERCA2b mRNA but not IP3R1 mRNA in cerebellar neurons may be an important mechanism underlying alteration of calcium homeostasis and cerebellar degeneration upon chronic ethanol consumption.

Administration, Oral↗

Effects of ischemic tolerance on mRNA levels of IP3R1, beta-actin, and neuron-specific enolase in hippocampal CA1 area of the gerbil brain.

Global cerebral ischemia induced to Mongolian gerbils by ligation of common carotid arteries (CCAs) is known to result in injury to the hippocampal CA1 region. In this study, we examined whether neuronal injury can be depicted by measuring levels of mRNA encoding inositol 1,4,5-trisphosphate receptor type 1 (IP3R1), neuron specific enolase (NSE) and beta-actin and whether these measurements can be use to assess ischemic tolerance. Gerbils were subjected either to cerebral ischemia induced by ligation of both CCAs for 5 min, or to an ischemic tolerance paradigm in which a 2 min ischemic preconditioning was performed 24 hr prior to the 5 min ischemia. At 48 hr after the 5 min ischemic insult, significant decreases in mRNA levels for IP3R1 (26%), NSE (38%) and beta-actin (50%) could be observed in the hippocampal CA1 region. Although levels of mRNA in the preconditioning group were decreased as compared to the sham control, the levels were significantly higher than those in the ischemic group. These results indicate the feasibility of using mRNA measurement as a parameter to assess cerebral ischemic damage. In addition, based on the differences in the decline in mRNA levels between the ischemia group and the preconditioned ischemia group, it can be concluded that this ischemic tolerance paradigm could offer partial protection (around 45%) against the injury due to the 5 min cerebral ischemic insult.

Actins↗

[Research progress on regulation of vascular endothelial barrier function].

The regulatory mechanisms of vascular endothelial barrier function are complicated. Inflammatory mediators, such as alpha-thrombin, activate phospholipases to mediate generation of IP3 and other second messengers through receptor-coupled G protein. Protein kinase C and myosin light chain kinase are then activated, leading to phosphorylation of myosin light chains, rearrangement of F-actin skeleton, interendothelial cell gap formation and increased endothelial permeability.

Animals↗

Prenatal ethanol exposure selectively reduces the mRNA encoding alpha-1 thyroid hormone receptor in fetal rat brain.

Some of the developmental defects characteristic of congenital or experimental hypothyroidism are also observed in children or experimental animals prenatally exposed to ethanol, suggesting that a subset of neurological defects attributable to ethanol exposure are produced by interfering with thyroid hormone action. In this article, we tested whether an ethanol treatment regimen known to produce neurological damage in rats can alter the expression of the mRNAs encoding the thyroid hormone receptor isoforms (TR alpha-1, TR alpha-2, and TR beta-1) in the fetal rat brain neocortex and hippocampus. Rats were fed an ethanol-containing diet beginning on gestational day (G) 6 and continuing until sacrifice on G15, G17, or G21; controls included animals pair-fed a liquid control diet or fed lab chow. Ethanol selectively reduced the expression of TR alpha-1 mRNA in the neocortex and hippocampus on G21, compared with pair-fed and control fetuses. In contrast, pair-feeding selectively reduced TR alpha-2 mRNA in both neocortex and hippocampus on G21, and increased TR beta-1 mRNA on G17. These data support the hypothesis that ethanol may interfere with thyroid hormone action during fetal brain development. In addition, these data indicate that ethanol and pair-feeding exert independent effects on thyroid hormone receptor expression in the developing brain.

Animals↗

Induction of basic fibroblast growth factor (bFGF) expression following focal cerebral ischemia.

Basic fibroblast growth factor (bFGF) is a biologically active polypeptide with mitogenic, angiogenic, and neurotrophic properties. In the present study, we examined the temporal and spatial expression profiles of bFGF mRNA and protein concentration in a focal cerebral ischemia model induced by transient occlusion of the right middle cerebral artery (MCA) and both common carotid arteries (CCAs). Results of Northern blot analysis shows a transient 2.5-fold increase in the 6.0 kb transcript of bFGF mRNA within the ischemic cortex of rats subjected to 60 min ischemic insult followed by 12 h of reperfusion. Although enhanced expression of bFGF mRNA was also noted in the ipsilateral hippocampus, the temporal induction profile appeared to be different from that of the ischemic cortex. A significant increase in bFGF mRNA was observed as early as 60 min following ischemia and remained elevated for up to 2 weeks after the onset of reperfusion. In situ hybridization studies revealed constitutive expression of bFGF mRNA in discrete brain regions of sham-operated animals. Following 60 min ischemia and 12 h reperfusion, increased expression of bFGF mRNA was observed in the ischemic cortex (both peri-infarct and infarct area). Increased expression of bFGF mRNA within the infarcted area is largely confined rostrally to the outer cortical layers of the infarct, an area with increased density of blood vessels. bFGF-like immunoreactivity was also detected in areas expressing bFGF mRNA. Furthermore, a striking increase in bFGF-like immunoreactivity was observed in the ipsilateral hippocampus. Double-staining with anti-GFAP antibody indicated that the majority of the bFGF-like immunoreactivity was localized in the astrocytes, however, not all astrocytes showed bFGF-like immunoreactivity. Some GFAP negative cell also showed bFGF-like immunoreactivity. In summary, increased expression of both bFGF mRNA and immunoreactivity following ischemia were located in the same brain regions. An increase in bFGF-like immunoreactivity after ischemic insult is likely due to an increase in the expression of its 6.0 kb bFGF mRNA transcripts. Although increased bFGF mRNA was observed in both ischemic cortex and ipsilateral hippocampus after ischemic insult, the temporal expression profiles differed. Results from the present study raise the possibility that increased expression of bFGF in the peri-infarcted area may limit the spread of ischemic injury.

Animals↗

An esterification protocol for cis-parinaric acid-determined lipid peroxidation in immune cells.

Loss of fluorescence from cis-parinaric acid (cPnA) is a sensitive indicator of lipid peroxidation. The purpose of this study was to utilize cPnA to determine, at the level of the intact immune cell, whether enrichment of membranes with polyunsaturated fatty acids (PUFA) increased lipid peroxidation. P388D1 macrophages were labeled by addition of cPnA as an ethanolic solution. Within two minutes of addition, in the absence-of serum, cPnA rapidly intercalated into the plasma membrane. Lipid peroxidation was initiated by addition of Fe(2+)-EDTA resulting in a dose-dependent decrease in fluorescence with increased oxidant concentration. Cells previously enriched with PUFA and labeled by intercalation showed no differences in spontaneous or Fe(2+)-induced lipid peroxidation. In separate experiments, 20 microM cPnA in ethanolic solution was injected into cell culture media containing 0.1% essentially fatty acid free bovine serum albumin (BSA). Cells were resuspended and incubated for 90 min at 37 degrees C. After washing with BSA to remove cPnA which had not incorporated, 0.5% (0.1 microM) of the added cPnA was found esterified within cellular lipids. This level of cPnA provided a 100-fold increase over basal autofluorescence levels. Cells labeled in this manner also lost fluorescence in a dose-dependent manner as levels of oxidant stress increased. Cells enriched with PUFA and labeled by esterification had significantly increased rates and total amounts of lipid peroxidation. Co-incubation with alpha-tocopherol and PUFA resulted in a decrease in lipid peroxidation which was not significantly different from control cells. In conclusion, esterification of cPnA into membrane phospholipids can sensitively detect changes in lipid peroxidation induced by alteration of membrane PUFA and/or vitamin E content.

Cells, Cultured↗

Membrane lipid metabolism and phospholipase activity in insect Spodoptera frugiperda 9 ovarian cells.

Although there is increasing use of insect ovarian Sf9 cells for the production of recombinant proteins, namely, via the baculovirus vector expression system, little is known about the lipids in the cell membrane and whether endogenous phospholipases are present for regulation of the cell membrane lipids. In this study, analysis of membrane lipids of Sf9 cells indicated the presence of phosphatidylethanolamine (PE) (diacyl type) and phosphatidylcholine as major phospholipids, followed by phosphatidylserine and phosphatidylinositol (PI), and only trace amounts of ethanolamine plasmalogen. These phospholipids contain high proportions of monoenoic fatty acids, e.g., 16:1 and 18:1, which comprise more than 70% of the total fatty acids although small amounts of polyunsaturated fatty acids such as 18:2 and 20:4 are also present. When Sf9 cells were incubated in a culture medium containing [14C]oleic acid and [14C]arachidonic acid, a large portion of the labels were incorporated into membrane phospholipids. Using [14C]arachidonoyl-phospholipids as substrates for incubation with cell homogenate and subcellular fractions, results indicate the presence of a ca(2+)-independent phospholipase A (PLA2) in the Sf9 cell cytosol fraction. This PLA2 shows a high preference for hydrolysis of PE and is active at a pH range of 7-9. Unlike the brain cells which contain active phospholipase C (PLC) specific for phosphatidylinositol, only limited amount of diacylglycerol (DAG) was released from [14C]arachidonoyl-PE in the Sf9 cells. Taken together, this study demonstrates active metabolism of membrane phospholipids in Sf9 cells, most likely mediated by acyltransferases and PLA2. Furthermore, despite the absence of PLC for PI, limited amount of DAG could be generated through hydrolysis of PE.

Animals↗

Effects of IL-1 beta on receptor-mediated poly-phosphoinositide signaling pathway in immortalized astrocytes (DITNC).

Astrocytes are known to play multi-functional roles in support of many homeostatic mechanisms in the central nervous system including host defense mechanisms. Despite the ability of cytokines to alter gene expression and cellular activity, their effect on receptor-mediated poly-phosphoinositide (poly-PI) signaling pathway has not been examined in detail. In this study, an immortalized astrocyte cell line (DITNC) was used to test the effect of IL-1 beta exposure on the poly-PI signaling pathway. Similar to primary astrocytes, DITNC cells exhibit P2-purinergic receptor response to ATP and UTP leading to transient increases in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and intracellular calcium concentration, [Ca2+]i. Upon exposure of DITNC cells to IL-1 beta (100 U/ml) for 24 hrs, an increased response to the poly-PI agonists was observed. The increase in ATP-mediated Ins(1,4,5)P3 release could not be attributed to a shift in the ATP dose or an alteration of the time profile for the release of Ins(1,4,5)P3. Since the increase in response required a lag time of 4 hr after IL-1 beta exposure, it is unlikely that this effect was due to a direct interaction of IL-1 beta with the purinergic receptor. On the other hand, an increase in ATP response could be observed in DITNC cells exposed to conditioned medium obtained after IL-1 beta treatment. It can be concluded that exposure of astrocytes to cytokines may lead to an increase in receptor-mediated poly-PI signaling activity and this may involve compounds secreted into the culture medium, e.g., the secretory phospholipase A2.

Adenosine Triphosphate↗

Chronic ethanol inhibits inositol metabolism in specific brain regions.

Many neurotransmitters and hormones in the nervous system transmit signals through receptors coupled to the poly-phosphoinositide (PI) signaling pathway. In this study, an in vivo protocol with [3H]inositol was used to examine the effect of chronic ethanol administration on inositol metabolism and poly-PI turnover in the cerebral cortex, hippocampus, and cerebellum of mouse brain. C57BL/6 mice were given a nutritionally complete liquid diet containing either ethanol (5%, w/v) or isocaloric sucrose for 2 months. Mice were injected intracerebrally with [3H]inositol; after 16 or 24 hr, they were injected intraperitoneally with lithium (8 mEq/kg body weight) to inhibit the inositol monophosphatase (IP1) activity. All mice were decapitated 4 hr after lithium injection. Labeled inositol phospholipids accounted for 16 to 23% of total labeled inositol in different regions of control mouse brain, and the percentages in the hippocampus were consistently higher than the cerebral cortex and cerebellum. In control mice, the percentages of labeled IP1 after a 4-hr lithium treatment were 11.5%, 9.9%, and 3.7% for cerebral cortex, hippocampus, and cerebellum, respectively. Chronic ethanol feeding resulted in a significant (p < 0.05) decrease in the percent of labeled IP1 and inositol phospholipids, and this effect was observed in the cerebral cortex and, to a lesser extent, hippocampus but not cerebellum. When ratios of labeled IP1 were expressed against labeled inositol phospholipids as an index of the poly-PI turnover activity, significant decreases in IP/lipid ratios were observed in the cerebral cortex, but not the hippocampus or cerebellum. Although mice killed 24 + 4 hr after the last ethanol feeding would have experienced an 8-hr period of ethanol withdrawal, compared with the 16 + 4-hr group, no differences in IP/lipid ratios were observed between the two time groups. These results illustrate regional differences in the effect of chronic ethanol on inositol metabolism in the brain, but no difference in poly-PI turnover in brain due to ethanol withdrawal.

Alcoholism↗

Expression of NGFI-B mRNA in a rat focal cerebral ischemia-reperfusion model.

Cerebral ischemia is known to induce the expression of several immediate early genes (IEGs), including c-fos and c-jun, which subsequently regulate a number of late effector genes. In this study, we examined the expression of NGFI-B (or nur 77) mRNA in a rat focal cerebral ischemia-reperfusion model. NGFI-B is a member of the IEGs which encodes for a nuclear receptor and is rapidly induced by nerve growth factor (NGF). Northern blot analysis showed a rapid but transient enhancement of NGFI-B mRNA, a peak level for which was observed at 30 min of reperfusion following 60 min ischemic insult. At the peak level, quantitative analysis of the blot indicated a 12-fold and 4-fold increase of NGFI-B mRNA in the ischemic cortex and ipsilateral hippocampus, respectively, as compared to the sham-operated control. No apparent changes in mRNA levels were observed within contralateral sites of the cortex. Results from in situ hybridization showed that severe ischemia (60 min) resulted in a marked increase of NGFI-B mRNA throughout the entire ischemic cerebral cortex. The increase was particularly notable in the frontal, occipital, perirhinal and piriform cortical regions and in the dentate gyrus and CAI-3 regions of the ipsilateral hippocampus. A marked induction was also noted in the ipsilateral caudate putamen. Unlike the induction profile of NGFI-B mRNA, severe ischemia resulted in bilateral increases of its family gene, NGFI-A mRNA. The spatial induction profile is similar to that of NGFI-B mRNA in both hemispheres, except within the region of the contralateral dentate gyrus which showed low levels of NGFI-A mRNA. The expression pattern of NGF and BDNF mRNA, upstream genes of NGFI-B, were also examined. Interestingly the temporal and spatial expression patterns of BDNF mRNA were very similar to that of NGFI-A mRNA under the same conditions, whereas increased NGF and NGFI-B mRNA were observed only in the ipsilateral hemisphere. It is likely that multiple and/or overlapping pathways are activated subsequent to ischemic challenge which in turn are crucial for cel survival and/or functional recovery following focal cerebral ischemia.

Analysis of Variance↗

Chronic ethanol on mRNA levels of IP3R1, IP3 3-kinase and mGluR1 in mouse Purkinje neurons.

Chronic ethanol exposure is known to alter the polyphosphoinositide (poly-PI) signaling pathway, although the exact sight of regulation is not well understood. In this study, C57B1/6J mice were given a control or an ethanol (6 g kg-1 body wt) liquid diet by gastric intubation daily for 4 weeks. Examination of levels of mRNA for type 1 inositol 1,4,5-trisphosphate receptor (IP3R1), inositol 1,4,5-trisphosphate 3-kinase (IP3K) and metabotropic glutamate receptor 1 (mGluR1) in cerebellar Purkinje neurons by in situ hybridization revealed significant decreases in cellular levels of mRNA encoding both IP3R1 and mGluR1 in the ethanol-treated group compared with controls, but levels of IP3K mRNA were not altered. These results suggest that chronic ethanol consumption selectively alters cellular mRNA in the poly-PI signaling pathway. The decrease in mRNA levels of mGluR1 and IP3R1 may be an important underlying mechanism towards explaining the alcohol-induced degenerative changes in the cerebellar Purkinje neurons.

Alcoholism↗

Poly-phosphoinositide-mediated messengers in focal cerebral ischemia and reperfusion.

The receptor-mediated poly-phosphoinositide (PI) signalling pathway is known to play an important role in maintaining intracellular calcium homeostasis, which in turn, is critical for mediating neuronal function. In this study, we examined the effects of focal cerebral ischemia induced in rats by temporary occlusion of the middle cerebral artery (MCA) and both common carotid arteries (CCAs) on this signal transduction pathway. Results indicate that several parts of the pathway are altered, both during the early phase of focal cerebral ischemic insult and after recirculation. Cerebral ischemia induced a decrease in levels of phosphatidylinositol 4,5-bisphosphate (PIP2) in the ischemic MCA cortex, due partly to stimulated poly-PI hydrolysis and partly to the depletion of ATP required for resynthesis of this substrate. ATP depletion during ischemia was also attributed to a sustained decrease in inositol 1,4,5-trisphosphate (IP3) levels. On the other hand, the decline in IP3 3-kinase activity after 30 min of ischemic insult was not related to ATP depletion. During reperfusion upon prolonged ischemic insult, neither IP3 level nor IP3 3-kinase activity were able to show recovery after reperfusion, despite that ATP levels recovered by 80%. In situ hybridization studies indicated a decrease in mRNA expression of IP3 receptor but not IP3 3-kinase during the initial 4 h of reperfusion after a 45 min ischemic insult. Under this same condition, insulted cortical neurons started to show morphological changes between 4 and 8 h after reperfusion and extensive cell death could be observed by 16 h. Taken together, these results demonstrated early and delayed changes in the poly-PI signalling pathway due to focal cerebral ischemia. These effects are likely to cause impairment in neuronal function and underline the process of cerebral ischemic damage.

Animals↗

Brain neutral lipids and phospholipids are modified by long-term feeding of beef tallow vs. corn oil diets.

Previous studies examining the response of brain lipids to dietary fat modification have not quantified neutral lipids such as diacylglycerols (DG) and triacylglycerols (TG). In this study we measured the concentrations of neutral lipids and phospholipids, and their fatty acid profiles, in the cerebra of rats fed defined diets containing either beef tallow (BT) or corn oil (CO) at 12% or 37% of energy. The diets were fed to rat pups beginning at 18 d of age and continued for 31 wk. The proportion of brain linoleic acid [18:2(n-6)] in TG from CO-fed rats was two- to fourfold greater than in BT-fed rats. Although 18:2(n-6) levels were higher in serum and brain TG of rats fed CO, differences in other TG fatty acid concentrations in serum were not reflected in the brain. Rats fed CO diets had higher concentrations of 18:2(n-6) in brain phospholipids as well as neutral lipids compared with rats fed BT diets, and the differences were greater in rats fed 37% rather than 12% of energy as fat. Differences in other polyunsaturated fatty acids associated with dietary fat composition were also found among the brain phospholipids. Most notably, the concentration of docosapentaenoic acid [22:5(n-6)] in brain phospholipids was highest in rats fed diets containing the lowest concentrations of alpha-linolenic acid [18:3(n-3)]. A concentration of 0.1 mg 18:3(n-3)/g diet appeared to be adequate to prevent elevation of 22:5(n-6) in brain phospholipids. These results demonstrate that consumption of a low fat diet (12% of energy) primarily comprised of saturated fats may potentiate an 18:3(n-3) deficiency in brain of rats.

Animals↗

Effects of ethanol on phosphorylation of lipids in rat synaptic plasma membranes.

Synaptic plasma membranes (SPM) isolated from rat cerebral cortex contain lipid kinases for conversion of phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), and diacylglycerol (DG) to PIP, phosphatidylinositol 4,5-bisphosphate (PIP2), and phosphatidic acid (PA), respectively. These anionic phospholipids are important in signal transduction mechanisms and are required for synaptic function. The effect of ethanol and other aliphatic alcohols on phosphorylation of these lipids in SPM has not been established. Incubation of SPM with [gamma-32P]ATP resulted in labeling of PIP, lyso-PIP, PIP2, and PA. Ethanol (50-200 mM) added to the incubation system showed a dose-dependent decrease in labeling of PIP2, but not PIP or PA. To a lesser extent, labeling of PIP2 was also inhibited by 1-propanol, but neither isopropanol nor 1-butanol could alter the PIP2 labeling pattern. Under similar incubation conditions, labeling of PIP and PA in SPM was not altered by ethanol, 1-propanol, iso-propanol, but 1-butanol stimulated PIP labeling with a peak at 25 mM. Addition of exogenous PIP to the incubation mixture led to an increase in labeling of PIP2, suggesting that the endogenous PIP pool in SPM is limiting for the synthesis of PIP2 in SPM. Interestingly, when SPM were incubated with exogenous PIP, addition of ethanol (50-100 mM) to this incubation mixture resulted in an increase in PIP2 labeling. Taken together, these results suggest a specific effect of ethanol on PIP kinase in SPM, and this effect seems to be dependent on the location and/or amount of PIP in the membrane.

Animals↗