Biomedical subjects

D J van der Horst

Publications and source records attributed to D J van der Horst.

At least 19 recordsLinked to original sources

PubMed2000

Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor from Locusta migratoria.

The transport of lipids via the circulatory system of animals constitutes a vital function that uses highly specialized lipoprotein complexes. In insects, a single lipoprotein, lipophorin, serves as a reusable shuttle for the transport of lipids between tissues. We have found that the two nonexchangeable apolipoproteins of lipophorin arise from a common precursor protein, apolipophorin II/I (apoLp-II/I). To examine the mechanisms of transport of lipids and liposoluble substances inside the central nervous system, this report provides the molecular cloning of a cDNA encoding the locust apoLp-II/I. We have recently shown that this precursor protein belongs to a superfamily of large lipid transfer proteins (Babin et al. [1999] J. Mol. Evol. 49:150-160). We determined that, in addition to its expression in the fat body, the locust apoLp-II/I is also expressed in the brain. Part of the signal resulted from fat body tissue associated with the brain; however, apoLp-II/I was strongly expressed and the corresponding protein detected, in pigmented glial cells of the lamina underlying the locust retina and in cells or cellular processes interspersed in the basement membrane. The latter finding strongly suggests an implication of apolipophorins in the transport of retinoids and/or fatty acids to the insect retina.

Amino Acid Sequence↗

PubMed2000

Lipid transport biochemistry and its role in energy production.

Recent advances on the biochemistry of flight-related lipid mobilization, transport, and metabolism are reviewed. The synthesis and release of adipokinetic hormones and their function in activation of fat body triacylglycerol lipase to produce diacylglycerol is discussed. The dynamics of reversible lipoprotein conversions and the structural properties and role of the exchangeable apolipoprotein, apolipophorin III, in this process is presented. The nature and structure of hemolymph lipid transfer particle and the potential role of a recently discovered lipoprotein receptor of the low-density lipoprotein receptor family, in lipophorin metabolism and lipid transport is reviewed.

Animals↗

PubMed1999

Adipokinetic hormones. Coupling between biosynthesis and release.

During long-distance flight of migratory locusts, the dramatic energy demand of the flight muscles is controlled by three adipokinetic hormones (AKHs). These peptide hormones regulate the mobilization of lipid and carbohydrate stored in the fat body to serve as energy substrates for the flight muscles. Despite the relatively huge quantities of the three AKHs that are stored in the corpora cardiaca, flight induces a differential 2-4-fold increase in the mRNAs for the three hormones. Moreover, newly synthesized AKHs can be released only during a restricted period of time, suggesting that by far most of the stored hormones are physiologically inactive. This raises the question of how the biosynthetic activity in the AKH-producing cells is coupled to their secretory activity. The present review discusses the potential mechanisms by which generation and release of mixtures of bioactive neurohormones are controlled and how peptidergic neuroendocrine cells cope with variations in physiological stimulation, with the AKH-producing cells serving as a model system.

Amino Acid Sequence↗

PubMed1995

The adipokinetic cells in the corpus cardiacum of Locusta migratoria preferentially release young secretory granules.

The influence of flight activity on the release of secretory granules from the adipokinetic cells in the corpus cardiacum of Locusta migratoria was studied. Two labeling methods, an enzymatical and a radioactive one, were used to label young, newly synthesized secretory granules and so distinguish them from older, preexisting granules. Both methods demonstrated that the ratio between the numbers of labeled and unlabeled secretory granules was lower in flight-stimulated adipokinetic cells than in unstimulated cells. This ratio was lower in both the cell bodies and the cell processes of flight-stimulated cells. After flight there was no detectable change in the total number of secretory granules, which indicates that the synthesis of new secretory granules is not inhibited by flight activity. Rather, the tendency of flight-stimulated cells to have more trans-Golgi networks labeled with wheat-germ agglutinin-conjugated horseradish peroxidase suggests that the synthesis of new secretory granules was enhanced by flight. The results led to the conclusion that young secretory granules were preferentially released over older secretory granules.

Amino Acids↗

PubMed1992

Biosynthesis and secretion of insect lipoprotein.

Biosynthesis of high density lipophorin (HDLp) was studied in larvae and adults of the migratory locust, Locusta migratoria. In an in vitro system, fat bodies were incubated in a medium containing a mixture of tritiated amino acids. Using SDS-PAGE and immunoblotting, it was shown that larval and adult fat bodies secreted both HDLp apoproteins, apolipophorin I (apoLp-I) and apolipophorin II (apoLp-II). Radiolabel was recovered in both apoproteins, indicative of de novo synthesis. The density of the fractions containing the apoproteins synthesized and secreted by larval and adult fat bodies was determined by density gradient ultracentrifugation. A radiolabeled protein fraction was found at density 1.12 g/ml. Using an enzyme-linked immunosorbent assay for detecting apoLp-I and apoLp-II, it was demonstrated that both apoproteins were present in this fraction, which had a density identical to that of circulating HDLp in hemolymph. Lipid analysis revealed that it contained phospholipid, diacylglycerol, sterol, and hydrocarbons. From these results it is concluded that the fat body of the locust synthesizes both apoLp-I and apoLp-II, which are combined with lipids to a lipoprotein particle that is released into the medium as HDLp.

Animals↗

PubMed1991

Lipophorin structure analyzed by in vitro treatment with lipases.

Adult Manduca sexta high density lipophorin (HDLp-A) is composed of three apolipoproteins (apoLp-I, -II, and -III) and 52% lipid. The flight-specific low density lipophorin (LDLp) contains 62% lipid and is associated with several additional molecules of apoLp-III. The amount of phospholipid remains constant in lipophorin (140 mol/mol of lipophorin), while the diacylglycerol content varies between different lipophorin species (310 mol/mol HDLp up to 1160 mol/mol LDLp). Both lipophorin particles were enzymatically depleted of phospholipid or diacylglycerol by in vitro incubation with either phospholipase A2 or triacylglycerol lipase. Albumin was used to remove free fatty acids generated during the reaction. Treatment with phospholipase A2 removed all phospholipids (except sphingomyelin) and the resulting particles were stable. Triacylglycerol lipase hydrolyzed large fractions of diacylglycerol. The resulting particles were smaller in size, higher in density, and devoid of apoLp-III. The particles retained apoLp-I and -II and the other lipid components, including a substantial amount of diacylglycerol. Structural integrity of diacylglycerol-depleted lipophorin was confirmed by electron microscopical analysis. When treated with both phospholipase A2 and triacylglycerol lipase, lipophorin precipitated. From these results we conclude that: 1) all phospholipid and apoLp-III are located at the surface of lipophorin, whereas diacylglycerol is partitioned between the sublayers and the surface of the particle; 2) both diacylglycerol and phospholipid play a role in stabilizing lipophorin in the aqueous medium; and 3) lipophorin can be extensively unloaded and still retain its basic structure, a necessary feature for its function as a reusable lipid shuttle.

Animals↗

PubMed1991

In vivo and in vitro loading of lipid by artificially lipid-depleted lipophorins: evidence for the role of lipophorin as a reusable lipid shuttle.

Lipid transport in the hemolymph of Manduca sexta is facilitated by a high density lipophorin in the resting adult insect (HDLp-A, d approximately 1.109 g/ml) and by a low density lipophorin during flight (LDLp, d approximately 1.060 g/ml). Lipophorin presumably shuttles different lipids between sites of uptake or storage, and sites of utilization. In order to shuttle lipid, a lipid-depleted lipophorin should be able to reload with lipid. To test this hypothesis, we used HDLp-A particles that were artificially depleted of either phospholipid (d approximately 1.118 g/ml) or diacylglycerol (d approximately 1.187 g/ml) and subsequently radiolabeled in their protein moiety. Upon injection into adult moths, both particles shifted their density to that of native HDLp-A, indicating lipid loading. Also, upon subsequent injection of adipokinetic hormone, both particles shifted to a lower density (d approximately 1.060 g/ml) indicating diacylglycerol loading and conversion to LDLp. Both phospholipid and diacylglycerol loading were also studied using an in vitro system. The lipid-depleted particles were incubated with fat body that had been radiolabeled in either the phospholipid or the triacylglycerol fraction. Transfer of radiolabeled phospholipid and diacylglycerol from fat body to lipophorin was observed. During diacylglycerol loading, apoLp-III associated with lipophorin, whereas phospholipid loading occurred in the absence of apoLp-III. The results show the ability of lipid-depleted lipophorins to reload with lipid and therefore reaffirm the role of lipophorin as a reusable lipid shuttle.

Animals↗

PubMed1987

Partial purification of locust flight muscle lipoprotein lipase (LpL): apparent differences from mammalian LpL.

1. An attempt was made to purify lipoprotein lipase (LpL) from the flight muscle of the migratory locust based on affinity for heparin, which is known to avidly bind mammalian LpL. 2. However, locust LpL appeared to completely lack this property, which indicates that the suggested membrane-binding of locust LpL is very different from that of mammalian LpL: a heparin-like glycosaminoglycan is not involved. 3. Since locust LpL lacks heparin affinity, other purification methods were assayed. Solubilization of locust LpL was obtained by the detergent Tween 20. 4. Though both anion and cation exchange chromatography resulted in the complete loss of enzyme activity, partial purification of locust LpL was achieved by gel filtration chromatography.

Animals↗

PubMed1980

Chiral 1,2-diacylglycerols in the haemolymph of the locust, Locusta migratoria.

A 1H-NMR method using chiral shift reagents was applied in the stereochemical analysis of the haemolymph 1,2-diacylglycerols of Locusta migratoria. Conversion of the 1,2-diacylglycerols into 1,2-diacetyl-3-tritylglycerols allowed the accurate determination of the enantiomeric purity, whereas direct trimethylsilylation of the unmodified or hydrogenated haemolymph 1,2-diacylglycerols proved to be less suitable because of signal broadening. In the haemolymph of Locusta, sn-1,2-diacylglycerols with a remarkably high optical purity were found to be present. In the resting locust, at least 96% of the haemolymph 1,2-diacylglycerols have the sn-1,2-configuration, in locusts in which the haemolymph diacylglycerol concentration was elevated by fat body triacylglycerol mobilization induced by flight activity or injection of adipokinetic hormone, over 97% of the 1,2-diacylglycerols is the sn-1,2-enantiomer. The few percent sn-2,3-enantiomer may not have been present initially. Positional distribution of the fatty acids in the fat body triacylglycerols and in the haemolymph sn-1,2-diacylglycerols obtained from locusts after a 2 h flight revealed nearly identical occupation of the sn-2-positions in both acylglycerols. The distribution patterns in the sn-1-position of the 1,2-diacylglycerols and the combined sn-1 and sn-3 positions of the triacylglycerols are compatible with the possible existence of a stereospecific sn-3-triacylglycerol lipase.

Animals↗

PubMed1976