Embryonic Lethality of Mitochondrial Pyruvate Carrier 1 Deficient Mouse Can Be Rescued by a Ketogenic Diet

Mitochondrial import of pyruvate by the mitochondrial pyruvate carrier (MPC) is a central step which links cytosolic and mitochondrial intermediary metabolism. To investigate the role of the MPC in mammalian physiology and development, we generated a mouse strain with complete loss of MPC1 expression. This resulted in embryonic lethality at around E13.5. Mouse embryonic fibroblasts (MEFs) derived from mutant mice displayed defective pyruvate-driven respiration as well as perturbed metabolic profiles, and both defects could be restored by reexpression of MPC1. Labeling experiments using ¹³C-labeled glucose and glutamine demonstrated that MPC deficiency causes increased glutaminolysis and reduced contribution of glucose-derived pyruvate to the TCA cycle. Morphological defects were observed in mutant embryonic brains, together with major alterations of their metabolome including lactic acidosis, diminished TCA cycle intermediates, energy deficit and a perturbed balance of neurotransmitters. Strikingly, these changes were reversed when the pregnant dams were fed a ketogenic diet, which provides acetyl-CoA directly to the TCA cycle and bypasses the need for a functional MPC. This allowed the normal gestation and development of MPC deficient pups, even though they all died within a few minutes post-delivery. This study establishes the MPC as a key player in regulating the metabolic state necessary for embryonic development, neurotransmitter balance and post-natal survival.     

Study of Plasmodium falciparum DHHC palmitoyl transferases identifies a role for PfDHHC9 in gametocytogenesis

Palmitoylation is the post‐translational reversible addition of the acyl moiety, palmitate, to cysteine residues of proteins and is involved in regulating protein trafficking, localization, stability and function. The Aspartate‐Histidine‐Histidine‐Cysteine (DHHC) protein family, named for their highly conserved DHHC signature motif, is thought to be responsible for catalysing protein palmitoylation. Palmitoylation is widespread in all eukaryotes, including the malaria parasite, Plasmodium falciparum, where over 400 palmitoylated proteins are present in the asexual intraerythrocytic schizont stage parasites, including proteins involved in key aspects of parasite maturation and development. The P. falciparum genome includes 12 proteins containing the conserved DHHC motif. In this study, we adapted a palmitoyl‐transferase activity assay for use with P. falciparum proteins and demonstrated for the first time that P. falciparum DHHC proteins are responsible for the palmitoylation of P. falciparum substrates. This assay also reveals that multiple DHHCs are capable of palmitoylating the same substrate, indicating functional redundancy at least in vitro. To test whether functional redundancy also exists in vivo, we investigated the endogenous localization and essentiality of a subset of schizont‐expressed PfDHHC proteins. Individual PfDHHC proteins localized to distinct organelles, including parasite‐specific organelles such as the rhoptries and inner membrane complex. Knock‐out studies identified individual DHHCs that may be essential for blood‐stage growth and others that were functionally redundant in the blood stages but may have functions in other stages of parasite development. Supporting this hypothesis, disruption of PfDHHC9 had no effect on blood‐stage growth but reduced the formation of gametocytes, suggesting that this protein could be exploited as a transmission‐blocking target. The localization and stage‐specific expression of the DHHC proteins may be important for regulating their substrate specificity and thus may provide a path for inhibitor development.     

Land use,rangeland degradation and ecological changes in the southern Kalahari,Botswana

Dual‐scale analyses assessing farm‐scale patterns of ecological change and landscape‐scale patterns of change in vegetation cover and animal distribution are presented from ecological transect studies away from waterpoints, regional remotely sensed analysis of vegetation cover and animal numbers across the southern Kalahari, Botswana. Bush encroachment is prevalent in semi‐arid sites where Acacia mellifera Benth. is widespread in communal areas and private ranches, showing that land tenure changes over the last 40 years have not avoided rangeland degradation. Herbaceous cover is dominated in intensively grazed areas by the annual grass Schmidtia kalahariensis Stent and in moderately grazed areas by the perennial grass Eragrostis lehmanniana Nees. Nutritious perennial grass species including Eragrostis pallens Hack. Ex Schinz remain prevalent in Wildlife Management Areas. Other ecological changes include the invasion of the exotic Prosopis glandulosa Torr. and dense stands of Rhigozum trichotomum Kuntze. in the arid southwest. Regional patterns of wildlife species show that the expansion of cattleposts and fenced ranches has led to large areas of low wildlife conservation value even in areas where cattle production is not practiced. Findings show the need for integrated landscape‐scale planning of land use if the ecological value and biodiversity of the southern Kalahari is to be retained.     

Identification of a microRNA (miR-663a) induced by ER stress and its target gene PLOD3 by a combined microRNome and proteome approach

Introduction

MicroRNAs (miRs) regulate gene expression to support important physiological functions. Significant evidences suggest that miRs play a crucial role in many pathological events and in the cell response to various stresses.

Methods

With the aim to identify new miRs induced by perturbation of intracellular calcium homeostasis, we analysed miR expression profiles of thapsigargin (TG)-treated cells by microarray. In order to identify miR-663a-regulated genes, we evaluated proteomic changes in miR-663a-overexpressing cells by two-dimensional differential in-gel electrophoresis coupled to mass spectrometric identification of the differentially represented proteins. Microarray and proteomic analyses were supported by biochemical validation.

Results

Results of microarray revealed 24 differentially expressed miRs; among them, miR-663a turned out to be by ER stress and under the control of the PERK pathway of the unfolded protein response. Proteomic analysis revealed that PLOD3, which is the gene encoding for collagen-modifying lysyl hydroxylase 3 (LH3), is regulated by miR-663a. Luciferase reporter assays demonstrated that miR-663a indeed reduces LH3 expression by targeting to 3′-UTR of PLOD3 mRNA. Interestingly, miR-663a inhibition of LH3 expression generates reduced extracellular accumulation of type IV collagen, thus suggesting the involvement of miR-663a in modulating collagen 4 secretion in physiological conditions and in response to ER stress.

Conclusion

The finding of the ER stress-induced PERK-miR-663a pathway may have important implications in the understanding of the molecular mechanisms underlying the function of this miR in normal and/or pathological conditions.

     

Softening the agricultural matrix: a novel agri‐environment scheme that balances habitat restoration and livestock grazing

The loss and degradation of woody vegetation in the agricultural matrix represents a key threat to biodiversity. Strategies for habitat restoration in these landscapes should maximize the biodiversity benefit for each dollar spent in order to achieve the greatest conservation outcomes with scarce funding. To be effective at scale, such strategies also need to account for the opportunity cost of restoration to the farmer. Here, we critique the Whole‐of‐Paddock Rehabilitation program, a novel agri‐environment scheme which seeks to provide a cost‐effective strategy for balancing habitat restoration and livestock grazing. The scheme involves the revegetation of large (minimum 10 ha) fields, designed to maximize biodiversity benefits and minimize costs while allowing for continued agricultural production. The objectives and design of the scheme are outlined, biodiversity and production benefits are discussed, and we contrast its cost‐effectiveness with alternative habitat restoration strategies. Our analysis indicates that this scheme achieves greater restoration outcomes at approximately half the cost of windbreak‐style plantings, the prevailing planting configuration in southeastern Australia, largely due to a focus on larger fields, and the avoidance of fencing costs through the use of existing farm configuration and infrastructure. This emphasis on cost‐effectiveness, the offsetting of opportunity costs through incentive payments, and the use of a planting design that seeks to maximize biodiversity benefits while achieving production benefits to the farmer, has the potential to achieve conservation in productive parts of the agricultural landscape that have traditionally been “off limits” to conservation.