A Type-I Diacylglycerol Acyltransferase Modulates Triacylglycerol Biosynthesis and Fatty Acid Composition in the Oleaginous Microalga, Nannochloropsis Oceanica

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Description

Background: Photosynthetic oleaginous microalgae are considered promising feedstocks for biofuels. The marine microalga, Nannochloropsis oceanica, has been attracting ever-increasing interest because of its fast growth, high triacylglycerol (TAG) content, and available genome sequence and genetic tools. Diacylglycerol acyltransferase (DGAT) catalyzes the

Background: Photosynthetic oleaginous microalgae are considered promising feedstocks for biofuels. The marine microalga, Nannochloropsis oceanica, has been attracting ever-increasing interest because of its fast growth, high triacylglycerol (TAG) content, and available genome sequence and genetic tools. Diacylglycerol acyltransferase (DGAT) catalyzes the last and committed step of TAG biosynthesis in the acyl-CoA-dependent pathway. Previous studies have identified 13 putative DGAT-encoding genes in the genome of N. oceanica, but the functional role of DGAT genes, especially type-I DGAT (DGAT1), remains ambiguous.

Results: Nannochloropsis oceanica IMET1 possesses two DGAT1 genes: NoDGAT1A and NoDGAT1B. Functional complementation demonstrated the capability of NoDGAT1A rather than NoDGAT1B to restore TAG synthesis in a TAG-deficient yeast strain. In vitro DGAT assays revealed that NoDGAT1A preferred saturated/monounsaturated acyl-CoAs and eukaryotic diacylglycerols (DAGs) for TAG synthesis, while NoDGAT1B had no detectable enzymatic activity. Assisted with green fluorescence protein (GFP) fusion, fluorescence microscopy analysis indicated the localization of NoDGAT1A in the chloroplast endoplasmic reticulum (cER) of N. oceanica. NoDGAT1A knockdown caused ~25% decline in TAG content upon nitrogen depletion, accompanied by the reduced C16:0, C18:0, and C18:1 in TAG sn-1/sn-3 positions and C18:1 in the TAG sn-2 position. NoDGAT1A overexpression, on the other hand, led to ~39% increase in TAG content upon nitrogen depletion, accompanied by the enhanced C16:0 and C18:1 in the TAG sn-1/sn-3 positions and C18:1 in the TAG sn-2 position. Interestingly, NoDGAT1A overexpression also promoted TAG accumulation (by ~2.4-fold) under nitrogen-replete conditions without compromising cell growth, and TAG yield of the overexpression line reached 0.49 g L[superscript −1] at the end of a 10-day batch culture, 47% greater than that of the control line.

Conclusions: Taken together, our work demonstrates the functional role of NoDGAT1A and sheds light on the underlying mechanism for the biosynthesis of various TAG species in N. oceanica. NoDGAT1A resides likely in cER and prefers to transfer C16 and C18 saturated/monounsaturated fatty acids to eukaryotic DAGs for TAG assembly. This work also provides insights into the rational genetic engineering of microalgae by manipulating rate-limiting enzymes such as DGAT to modulate TAG biosynthesis and fatty acid composition for biofuel production.

Date Created
2017-07-05
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Multisensory Interactions Influence Neuronal Spike Train Dynamics in the Posterior Parietal Cortex

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Although significant progress has been made in understanding multisensory interactions at the behavioral level, their underlying neural mechanisms remain relatively poorly understood in cortical areas, particularly during the control of action. In recent experiments where animals reached to and actively

Although significant progress has been made in understanding multisensory interactions at the behavioral level, their underlying neural mechanisms remain relatively poorly understood in cortical areas, particularly during the control of action. In recent experiments where animals reached to and actively maintained their arm position at multiple spatial locations while receiving either proprioceptive or visual-proprioceptive position feedback, multisensory interactions were shown to be associated with reduced spiking (i.e. subadditivity) as well as reduced intra-trial and across-trial spiking variability in the superior parietal lobule (SPL). To further explore the nature of such interaction-induced changes in spiking variability we quantified the spike train dynamics of 231 of these neurons. Neurons were classified as Poisson, bursty, refractory, or oscillatory (in the 13–30 Hz “beta-band”) based on their spike train power spectra and autocorrelograms. No neurons were classified as Poisson-like in either the proprioceptive or visual-proprioceptive conditions. Instead, oscillatory spiking was most commonly observed with many neurons exhibiting these oscillations under only one set of feedback conditions. The results suggest that the SPL may belong to a putative beta-synchronized network for arm position maintenance and that position estimation may be subserved by different subsets of neurons within this network depending on available sensory information. In addition, the nature of the observed spiking variability suggests that models of multisensory interactions in the SPL should account for both Poisson-like and non-Poisson variability.

Date Created
2016-12-29
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Limb position estimation: neural mechanisms and consequences for movement production

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Description
An accurate sense of upper limb position is crucial to reaching movements where sensory information about upper limb position and target location is combined to specify critical features of the movement plan. This dissertation was dedicated to studying the mechanisms

An accurate sense of upper limb position is crucial to reaching movements where sensory information about upper limb position and target location is combined to specify critical features of the movement plan. This dissertation was dedicated to studying the mechanisms of how the brain estimates the limb position in space and the consequences of misestimation of limb position on movements. Two independent but related studies were performed. The first involved characterizing the neural mechanisms of limb position estimation in the non-human primate brain. Single unit recordings were obtained in area 5 of the posterior parietal cortex in order to examine the role of this area in estimating limb position based on visual and somatic signals (proprioceptive, efference copy). When examined individually, many area 5 neurons were tuned to the position of the limb in the workspace but very few neurons were modulated by visual feedback. At the population level however decoding of limb position was somewhat more accurate when visual feedback was provided. These findings support a role for area 5 in limb position estimation but also suggest that visual signals regarding limb position are only weakly represented in this area, and only at the population level. The second part of this dissertation focused on the consequences of misestimation of limb position for movement production. It is well known that limb movements are inherently variable. This variability could be the result of noise arising at one or more stages of movement production. Here we used biomechanical modeling and simulation techniques to characterize movement variability resulting from noise in estimating limb position ('sensing noise') and in planning required movement vectors ('planning noise'), and compared that to the variability expected due to noise in movement execution. We found that the effects of sensing and planning related noise on movement variability were dependent upon both the planned movement direction and the initial configuration of the arm and were different in many respects from the effects of execution noise.
Date Created
2011
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