RESUMEN
PURPOSE: In a previous study, it was reported that docosahexaenoic acid (DHA) is essential to postpone apoptosis and to promote differentiation of rat retina photoreceptors in vitro. In the current study, the protective effects of GDNF on photoreceptor cells during development in vitro and its action when combined with DHA were investigated. METHODS: Rat retina neuronal cultures were incubated in a chemically defined medium, either without photoreceptor survival factors or supplemented with GDNF, DHA, or GDNF plus DHA. Evolution of survival, apoptosis, opsin expression, mitochondrial functioning, and cell proliferation were investigated at different times of development in vitro. RESULTS: Incubation with GDNF selectively increased the number of surviving photoreceptors, reduced their apoptosis, and augmented opsin expression. Proliferative cell nuclei antigen (PCNA) determination and addition of [(3)H]-thymidine or bromodeoxyuridine showed that GDNF promoted neuroblast proliferation during the first hours of development in vitro. The combined addition of GDNF and DHA enhanced opsin expression and photoreceptor survival in an additive manner. The advance of photoreceptor apoptosis in cultures without trophic factors correlated with an increased impairment in mitochondrial functionality. Addition of GDNF and DHA significantly diminished the loss of mitochondrial activity. CONCLUSIONS: These results show that GDNF stimulated the cell cycle progression, leading to neuroblast proliferation at early stages of development, and delayed the onset of apoptosis later on, improving differentiation and acting as a trophic factor for photoreceptors. The combination of GDNF with DHA had an additive effect both on photoreceptor survival and on opsin expression. Preservation of mitochondrial function may be involved in the antiapoptotic effect of both factors.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Factores de Crecimiento Nervioso , Proteínas del Tejido Nervioso/farmacología , Células Fotorreceptoras de Vertebrados/citología , Animales , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ciclina G , Ciclina G1 , Ciclinas/metabolismo , ADN/biosíntesis , Combinación de Medicamentos , Sinergismo Farmacológico , Factor Neurotrófico Derivado de la Línea Celular Glial , Mitocondrias/fisiología , Células Fotorreceptoras de Vertebrados/efectos de los fármacos , Células Fotorreceptoras de Vertebrados/metabolismo , Antígeno Nuclear de Célula en Proliferación/metabolismo , Ratas , Ratas Wistar , Opsinas de Bastones/metabolismoRESUMEN
In this study we show that insulin-like growth factor (IGF)-I selectively promotes survival and differentiation of amacrine neurons. In cultures lacking this factor, an initial degeneration pathway, selectively affecting amacrine neurons, led to no lamellipodia development and little axon outgrowth. Cell lysis initially affected 50% of amacrine neurons; those remaining underwent apoptosis leading to the death of approximately 95% of them by day 10. Apoptosis was preceded by a marked increase in c-Jun expression. Addition of IGF-I or high concentrations (over 1 microM) of either insulin or IGF-II to the cultures prevented the degeneration of amacrine neurons, stimulated their neurite outgrowth, increased phospho-Akt expression and decreased c-Jun expression. The high insulin and IGF-II concentrations required to protect amacrine cells suggest that these neurons depend on IGF-I for their survival, IGF-II and insulin probably acting through IGF-I receptors to mimic IGF-I effects. Inhibition of phosphatidylinositol-3 kinase (PI 3-kinase) with wortmannin blocked insulin-mediated survival. Wortmannin addition had similar effects to IGF-I deprivation: it prevented neurite outgrowth, increased c-Jun expression and induced apoptosis. These results suggest that IGF-I is essential for the survival and differentiation of amacrine neurons, and activation of PI 3-kinase is involved in the intracellular signaling pathways mediating these effects.
Asunto(s)
Factor I del Crecimiento Similar a la Insulina/metabolismo , Neuronas/metabolismo , Proteínas Serina-Treonina Quinasas , Retina/metabolismo , Androstadienos/farmacología , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Insulina/farmacología , Antagonistas de Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/farmacología , Factor II del Crecimiento Similar a la Insulina/metabolismo , Factor II del Crecimiento Similar a la Insulina/farmacología , Neuritas/efectos de los fármacos , Neuronas/citología , Neuronas/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-akt , Proteínas Proto-Oncogénicas c-jun/biosíntesis , Seudópodos/efectos de los fármacos , Ratas , Ratas Wistar , Retina/citología , Retina/efectos de los fármacos , WortmaninaAsunto(s)
Ácidos Docosahexaenoicos/farmacología , Ácidos Grasos Esenciales/farmacología , Células Fotorreceptoras de Vertebrados/citología , Células Fotorreceptoras de Vertebrados/fisiología , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Células Cultivadas , Neuronas/citología , Neuronas/efectos de los fármacos , Neuronas/fisiología , Células Fotorreceptoras de Vertebrados/efectos de los fármacos , Ratas , Retina/citología , Retina/fisiologíaRESUMEN
PURPOSE: The purpose of this work was to study the effects of diverse fatty acids on the composition, metabolism, differentiation, and characteristics of opsin expression in retina photoreceptors. METHODS: Cultures of rat retinal neurons were incubated with or without 22:6 n-3, 22:5 n-3, 20:4 n-6, 18:1 n-9, and 16:0, labeled and unlabeled. RESULTS: In photoreceptor cells incubated with 22:6 n-3 and 22:5 n-3, the proportions of these fatty acids in phospholipids increased four- to sixfold. The remaining fatty acids did not change lipid acyl chain composition. The labeled fatty acids were all actively esterified in neuronal lipids, particularly in phosphatidylcholine. Addition of unlabeled 22:6 n-3 did not affect the distribution among lipids of the other fatty acids but displaced [3H]20:4 n-6 from phosphatidylcholine and phosphatidylethanolamine. These results suggest that retinal neurons have specific mechanisms for processing fatty acids of different lengths and degrees of unsaturation and that 22:6 n-3 incorporation takes priority. Of all fatty acids, 22:6 n-3 was the most effective in promoting photoreceptor differentiation. In 22:6-sufficient photoreceptors, new apical processes formed, the expression of opsin augmented, and its localization improved, concentrating in the apical processes of the cells. CONCLUSIONS: The advancement in differentiation selectively elicited by 22:6 correlates with the fact that 22:6 n-3, but none of the other fatty acids, delays significantly the onset of apoptosis in photoreceptors in culture. The synthesis of 22:6-containing phospholipid molecules could be required for the proper localization of opsin. This could contribute to furthering the differentiation of photoreceptors, preventing their apoptosis, and extending their survival.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Neuronas/citología , Células Fotorreceptoras de Vertebrados/citología , Células Fotorreceptoras de Vertebrados/metabolismo , Animales , Animales Recién Nacidos , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Ácidos Grasos/metabolismo , Inmunohistoquímica , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fosfolípidos/metabolismo , Células Fotorreceptoras de Vertebrados/efectos de los fármacos , Ratas , Ratas Wistar , Opsinas de Bastones/metabolismoRESUMEN
When rat retinal cells are cultured in a serum-free medium, the photoreceptor cells start dying after 7 days. The addition of docosahexaenoic acid (DHA) to the cultures prevents the selective death of photoreceptors. Here it is shown that, unlike other retinal neurons, photoreceptors die through an apoptotic pathway. Hallmarks of apoptosis, such as nuclear fragmentation and condensation and DNA cleavage forming a ladder pattern on an agarose gel, were observed. The timing and high selectivity of the triggering of photoreceptor cell apoptosis suggest the existence of a programmed cell death. Compared with other fatty acids, DHA not only was the most effective in promoting photoreceptor survival, but also the only one to decrease the number of apoptotic nuclei. The results suggest that DHA is important among the factors preventing apoptosis of photoreceptors in the developing retina. A limitation in the availability of this fatty acid might trigger apoptosis as a result of the failure to develop functional photoreceptor outer segments.
Asunto(s)
Apoptosis , Ácidos Docosahexaenoicos/farmacología , Células Fotorreceptoras/fisiología , Retina/crecimiento & desarrollo , Animales , Núcleo Celular/ultraestructura , Células Cultivadas , Medio de Cultivo Libre de Suero , Fragmentación del ADN , Inmunohistoquímica , Células Fotorreceptoras/efectos de los fármacos , Células Fotorreceptoras/ultraestructura , Ratas , Ratas Wistar , Retina/citologíaRESUMEN
The formation of 14C-labelled long-chain and very-long-chain (n-3) pentaenoic and hexaenoic fatty acids was studied in bovine retina by following the metabolism of. [14C]-docosapentaenoate [C22:5, n-3 fatty acid (22:5 n-3)], [14C]-docosahexaenoate (22:6 n-3), and [14C]acetate. With similar amounts of 22:5 n-3 and 22:6 n-3 as substrates, the former was actively transformed into 24:5 n-3, whereas the latter was virtually unmodified. Labelled 24:5, 26:5, 24:6 and 22:6 were formed from [1-14C]22:5 n-3, showing that pentaenoic fatty acids including 24:5 n-3 can be elongated and desaturated within the retina. When retinal microsomes were incubated with [1-14C]22:5 n-3, 24:5 n-3 was the only fatty acid formed. In retinas incubated with [14C]acetate, 24:5 n-3 was the most highly labelled fatty acid among the polyenes synthesized, 24:6 n-3 being a minor product. Such selectivity in the elongation of two fatty acids identical in length, 22:5 n-3 and 22:6 n-3, despite the fact that 22:5 is a minor and 22:6 a major fatty acid constituent of retina, suggests that the active formation of 24:5 n-3 plays a key role in n-3 polyunsaturated fatty acid (PUFA) metabolism. This compound might give rise to even longer pentaenes via elongation, and to the major PUFAs of retina, 22:6 n-3, by 6-desaturation and chain shortening. Of all retinal lipids, a minor component, triacylglycerol (TG), incorporated the largest amounts of [14C]22:5 and 22:6. TG also concentrated most of the [14C]24:5 formed in retina, whether from [14C]22:5 n-3 or from [14C]acetate, suggesting an important role for this lipid in supporting PUFA metabolism and the synthesis of 22:6 n-3.
Asunto(s)
Ácidos Docosahexaenoicos/metabolismo , Ácidos Grasos Insaturados/metabolismo , Microsomas/metabolismo , Fosfolípidos/metabolismo , Retina/metabolismo , Acetatos/metabolismo , Animales , Transporte Biológico , Radioisótopos de Carbono , Bovinos , Ésteres del Colesterol/metabolismo , Diglicéridos/metabolismo , Ácidos Fosfatidicos/metabolismo , Fosfolípidos/aislamiento & purificación , Técnica de Dilución de Radioisótopos , Esfingomielinas/metabolismo , Triglicéridos/metabolismoRESUMEN
The effect of docosahexaenoic acid (DHA) on neuronal survival was studied in cultured cells isolated from newborn rat retina. In vivo, the content of DHA in the retina increased nearly fourfold from days 2 to 12 after birth, whereas in retinal cells in culture it remained constant. Unlike amacrine cells, the photoreceptor cells in control cultures underwent a selective degeneration, starting at day 7, that led to their massive death by day 11. The addition of DHA at day 7 led to its active incorporation by the cultures, increasing from 6 to 21% of total fatty acids in cell lipids, and completely prevented photoreceptor cell death. When other fatty acids were tested, both neuronal fatty acid composition and photoreceptor death were the same as in control cultures. These results indicate that DHA is specifically required for the survival of retinal photoreceptors.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Células Fotorreceptoras/efectos de los fármacos , Células Fotorreceptoras/fisiología , Retina/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Ácidos Docosahexaenoicos/farmacocinética , Ácidos Grasos/farmacología , Ratas , Ratas Wistar , Retina/citologíaRESUMEN
Clofibrate administration significantly altered the amount and fatty acid composition of lipids in mouse liver. The net content of phospholipids (PL) increased and that of triacylglycerols (TG) decreased concomitantly with liver enlargement in mice treated for two weeks with this drug (0.5% w/w in the food). The highest increase among PL was in phosphatidylcholine; other components either showed lower increases or, as in the case of sphingomyelin and the plasmalogens, decreased. In all lipid classes the treatment resulted in altered ratios between major saturates, between saturates and monoenes, and between major polyenes. Among these, 20:3n-6 and 22:5n-3 increased several-fold, and the 20:3n-6/20:4n-6 and 22:5n-3/22:6n-3 ratios increased due to a more active formation of the precursors than of the corresponding products. This change affected all glycerolipid classes. Liver sphingomyelin showed a relative enrichment in monoenoic fatty acids like 22:1 and 24:1, caused by a net decrease in the amount of saturates, particularly 22:0 and 24:0. The stimulated membrane proliferation imposed by clofibrate must increase phospholipid synthesis and, hence, the need for fatty acids. The results suggest that these demands are met mostly by TG acyl groups, either directly or after oxidation/desaturation processes. This was apparently the case for the polyenoic fatty acids of the n-6 and n-3 series. The longer chain (C22 and C24) components decreased, suggesting that their oxidation was stimulated to provide part of the required (C20 and C22) polyenes.
Asunto(s)
Clofibrato/farmacología , Ácidos Grasos/metabolismo , Hipolipemiantes/farmacología , Metabolismo de los Lípidos , Hígado/efectos de los fármacos , Hígado/metabolismo , Animales , Ácidos Grasos Omega-3/metabolismo , Ácidos Grasos Omega-6 , Ácidos Grasos Insaturados/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Fosfatidilcolinas/metabolismo , Fosfolípidos/metabolismo , Esfingomielinas/metabolismo , Triglicéridos/metabolismoRESUMEN
Dietary deficiency of essential fatty acids of the n-3 and n-6 series is known to promote a compensatory increase in polyenoic fatty acids of the n-9 series in the lipids of mammalian tissues. In the present study long-chain n-9 polyenes were found to be normal components of the epididymis and especially of sperm isolated from that tissue, in healthy, well-fed, fertile rats maintained on essential fatty acid-sufficient diets. The n-9 polyenes occurred in large concentrations in the choline glycerophospholipids (CGP), the major phospholipid class of spermatozoa in epididymal cauda, and were highly concentrated in plasmenylcholine, the major subclass of CGP. The uncommon polyene 22:4n-9 was found in the highest proportion, followed in order of relative abundance by 22:3n-9, 20:3n-9 and 24:4n-9. These polyenes were probably derived from oleate (18:1n-9) in much the same way as long-chain polyenes of the n-6 and n-3 series are derived from linoleate (18:2n-6) and linolenate (18:3n-3), respectively.
Asunto(s)
Epidídimo/química , Ácidos Grasos Insaturados/análisis , Espermatozoides/química , Animales , Dieta , Fertilidad/fisiología , Masculino , Modelos Biológicos , Fosfatidilcolinas/análisis , Fosfatidiletanolaminas/análisis , Plasmalógenos/análisis , Ratas , Ratas Endogámicas , Ratas Wistar , Distribución TisularRESUMEN
In their transit from the caput to the cauda segments of the epididymis, rat spermatozoa undergo significant modifications in lipid content and composition. The amount of lipid phosphorus per cell decreases, and most lipid classes show specific changes in their constituent fatty acids. A depletion of phosphatidylcholine and phosphatidylethanolamine, concomitant with a virtually unchanged amount of the corresponding plasmalogens, are the major alterations, plasmenylcholine thereby becoming the major phospholipid. Diphosphatidylglycerol, sphingomyelin and the phosphoinositides decrease to a lesser extent or do not change at all, also resulting in relative increases with sperm maturation. Concerning the fatty acids, the proportions of oleate (C18:1, n-9) and linoleate (C18:2, n-6) in most lipids decrease on movement of sperm from caput to cauda, augmenting in turn the proportions of longer-chain (C20 to C24) and more unsaturated fatty acids. Docosapentaenoate (C22:5, n-6) is a major acyl chain present in all lipids at both stages, but uncommon long-chain polyenoic fatty acids of the n-9 series are also present, being almost exclusively found in the choline glycerophospholipids. These fatty acids are found to undergo the most significant changes during sperm maturation. They are minor components of plasmenylcholine in immature spermatozoa, but increase severalfold on maturation, representing more than half of the acyl chains of this major lipid in cells from the cauda. The high concentration of n-9 polyenes in mature sperm plasmenylcholine raises intriguing questions on the possible role epididymal cells may play in providing spermatozoa with such an unusual phospholipid. These plasmenylcholines could contribute to the characteristic lipid domain organization of the mature spermatozoa plasma membrane.
Asunto(s)
Epidídimo/metabolismo , Ácidos Grasos Insaturados/metabolismo , Metabolismo de los Lípidos , Plasmalógenos/metabolismo , Espermatozoides/crecimiento & desarrollo , Animales , Epidídimo/crecimiento & desarrollo , Epidídimo/fisiología , Técnicas In Vitro , Masculino , Fosfatidilcolinas/metabolismo , Fosfolípidos/metabolismo , Ratas , Ratas Endogámicas , Maduración Sexual/fisiología , Espermatozoides/metabolismoRESUMEN
The synthesis of very long chain (C24 to C36) polyunsaturated (four, five and six double bonds) fatty acids (VLCPUFA) is investigated in bovine retina using [14C]acetate. Saturates on the one hand (mainly palmitate), and polyenes on the other (mainly VLCPUFA), incorporate most of the label found in lipids. Phosphatidylcholine (PC) is the most highly labelled lipid class, since both types of 14C-labelled fatty acids, but especially this novel series of VLCPUFA, are concentrated in this phospholipid. Radioactivity from [14C]acetate is found in very long chain tetra, penta and hexaenoic fatty acids of PC. The labelling of 20:4(n - 6), 20:5(n - 3), 22:5(n - 6) and 22:6(n - 3) is much lower than that of longer polyenes of each of these series, indicating that VLCPUFA are synthesized in situ by successive elongations of the above polyenes, pre-existing in retina lipids. In various subcellular fractions isolated from retinas after incubations with [14C]acetate (including cytosol, microsomes, mitochondria and photoreceptor membranes), the labelling of the VLCPUFA of PC is very high, even at relatively short intervals of incubation. The results suggest that not only the synthesis but also the intracellular traffic among membranes of VLCPUFA-containing species of PC are very active processes in the retina.
Asunto(s)
Ácidos Grasos Insaturados/biosíntesis , Retina/metabolismo , Acetatos/metabolismo , Animales , Bovinos , Metabolismo de los Lípidos , Fosfatidilcolinas/metabolismo , Fracciones Subcelulares/metabolismoRESUMEN
The in vivo labeling of electrocyte lipids is followed after injection of radioactive glycerol and two fatty acids, oleate and arachidonate, into the electric organ of an elasmobranch (Discopyge tschudii). De novo synthesis of lipids and acyl-exchange reactions are operative in the electrocyte. The three precursors are preferentially incorporated into phosphatidylcholine, phosphatidylinositol, and triacylglycerols. The highest specific activities are attained by triacylglycerols and polyphosphoinositides. Electrocyte stacks from electric organ show an efficient and continuous esterification of oleate and arachidonate into lipids after several hours of incubation. Except for an apparently more active labeling of triacylglycerols, which is attributed to the larger availability of free fatty acid precursors under the in vitro experimental conditions, the pattern of lipid labeling is similar to that attained in vivo. 32P-labeled lipids are also steadily produced in electrocyte stacks (24 h of incubation with [32P]phosphate) using glucose as the sole exogenous source of energy. Polyphosphoinositides are the lipids preferentially labeled. The ability to sustain the labeling of lipids under in vitro conditions renders isolated electrocyte stacks an interesting model for future research on lipid involvement in cholinergic function.
Asunto(s)
Pez Eléctrico/metabolismo , Órgano Eléctrico/metabolismo , Metabolismo de los Lípidos , Animales , Ácido Araquidónico , Ácidos Araquidónicos/metabolismo , Radioisótopos de Carbono , Esterificación , Glicerol/metabolismo , Cinética , Ácido Oléico , Ácidos Oléicos/metabolismo , Fosfatos/metabolismo , Ácidos Fosfatidicos/biosíntesis , Fosfatidilcolinas/biosíntesis , Fosfatidiletanolaminas/biosíntesis , Fosfatidilinositoles/biosíntesis , Fosfatidilserinas/biosíntesis , Triglicéridos/biosíntesis , TritioRESUMEN
The composition of phospholipids from electric organ and from membranes enriched in acetylcholine receptors (AChRs) is analyzed in three elasmobranch fish (Torpedo marmorata, Torpedo californica, and Discopyge tschudii). Irrespective of their purity, AChR-containing membranes are similar to electric organ in lipid and fatty acid composition. The following characteristics are common to the three species: (a) Choline, ethanolamine, and serine glycerophospholipids account for 80-90% of the phospholipids. (b) Their major fatty acid constituents are monoenes, saturates, and long-chain (n-3) polyenes (especially docosahexaenoate). (c) A large proportion of the ethanolamine glycerophospholipids (30-50%) is made up by plasmenylethanolamine, which contains fewer polyenes than phosphatidylethanolamine per mole of lipid. (d) Polyphosphoinositides represent 20-30% of the inositides of electric organ. (e) Phosphatidylinositol and phosphatidate have large proportions of 20- and 22-carbon polyenes. (f) Diphosphatidylglycerol and triacylglycerols are rich in oleate but also contain long-chain polyenes. (g) Sphingomyelin has monoenes and saturates ranging from 14 to 26 carbons. Species-related variations are observed (a) in the ratios between some phospholipid classes and subclasses and (b) in the relative abundance of the major polyunsaturated acyl chains of phospholipids. Despite these differences, the average unsaturation and length of fatty acids in major phospholipid classes are similar for the three species.
Asunto(s)
Membrana Celular/análisis , Pez Eléctrico/metabolismo , Órgano Eléctrico/análisis , Lípidos/análisis , Receptores Colinérgicos/metabolismo , Torpedo/metabolismo , Animales , Colesterol/análisis , Ácidos Grasos/análisis , Lípidos de la Membrana/análisis , Ácidos Fosfatidicos/análisis , Fosfatidilcolinas/análisis , Fosfatidiletanolaminas/análisis , Fosfatidilinositoles/análisis , Fosfatidilserinas/análisis , Especificidad de la Especie , Esfingomielinas/análisisRESUMEN
(1-14C)-labeled (n-6) eicosatetraenoate, (n-3) docosapentaenoate and (n-3) docosahexaenoate (20:4, 22:5 and 22:6, respectively) are efficiently taken up and actively esterified into the lipids of bovine retina after 2 h incubation. Photoreceptor membranes, mitochondria, microsomes and postmicrosomal supernatants, which display significant differences in phospholipid and fatty acid compositions, are isolated after such incubations to study the labeling of lipids. The lipid classes preferentially labeled with the acids (1) largely differ among and within subcellular fractions, while (2) some common features in the treatment of the three polyenes are observed in each fraction. In all of them, the three acids are actively incorporated in phosphatidylcholine; ethanolamine glycerophospholipid, phosphatidylserine (PS) and phosphatidylinositol (PI) are highly labeled with 22:6, 22:5 and 20:4 respectively; within ethanolamine glycerophospholipid, the three label phosphatidylethanolamine in preference to plasmenylethanolamine. Most of the 14C esterified in mitochondria is in phospholipids. The endoplasmic reticulum produces in addition highly labeled triacylglycerols, also found in cytosol. High levels of 14C-labeled diacylglycerols are observed exclusively in photoreceptor membranes, where the specific radioactivity of PI is very high. The total amounts of 14C incorporated (1) are in general similar within a given fraction for the three polyenes, but (2) largely differ among fractions. The labeling of the highly unsaturated phospholipids of photoreceptor membranes is the lowest, while the postmicrosomal supernatant (whose lipids are relatively the poorest in polyenoic fatty acids) contains most of the labeled lipids isolated from retinas under these conditions. The results indicate that polyunsaturated species of retina phospholipids undergo an active synthesis and turnover, as well as an intense intracellular traffic among membranes.
Asunto(s)
Ácidos Araquidónicos/metabolismo , Ácidos Docosahexaenoicos/metabolismo , Ácidos Grasos Insaturados/metabolismo , Lípidos de la Membrana/metabolismo , Retina/ultraestructura , Fracciones Subcelulares/metabolismo , Animales , Ácido Araquidónico , Bovinos , Retículo Endoplásmico/metabolismo , Esterificación , Microsomas/metabolismo , Mitocondrias/metabolismo , Fosfatidilcolinas/metabolismo , Fosfatidiletanolaminas/metabolismo , Fosfatidilinositoles/metabolismo , Fosfatidilserinas/metabolismo , Fosfolípidos/metabolismo , Segmento Externo de la Célula en Bastón/metabolismoRESUMEN
The labeling of molecular species of phosphatidylcholine (PC) has been studied in bovine retinas incubated for 2 h with (1-14C)-labeled (n-6) eicosatetraenoate (n-3) docosapentaenoate and (n-3) docosahexaenoate (20:4, 22:5 and 22:6, respectively) and in four subcellular fractions isolated after such incubations. Of the total radioactivity incorporated in PC, the following percentages of the above fatty acids, respectively, are found in its dipolyunsaturated species: 58, 56 and 53% in rod outer segments; 29, 41 and 49% in mitochondria; 24, 28 and 39% in microsomes; 12, 14 and 16% in postmicrosomal supernatants; 28, 36 and 58% in entire retinas. The remainder percentages are in tetra-, penta- and hexaenoic species of PC, respectively. The levels of pentaenoic species in the PCs of all fractions are similar, while tetraenes are lowest and hexaenes highest in photoreceptor membranes. Dipolyunsaturated species are highly concentrated in photoreceptor membranes, but are minor components of mitochondrial, microsomal and cytosolic PC. The specific radioactivities of tetraenoic, pentaenoic and hexaenoic PCs are decreasingly lower in the following order: postmicrosomal supernatants, microsomes, mitochondria, photoreceptor membranes. In contrast, the specific radioactivities of dipolyunsaturated PCs are higher in mitochondria and microsomes than in the other fractions, especially with 22:5 and 22:6. It is suggested that mitochondria as well as the endoplasmic reticulum could play a role in the synthesis and further modifications of dipolyunsaturated PCs before being supplied to photoreceptor membranes.