School of BioSciences - Research Publications

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Now showing 1 - 10 of 2008
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    Assessment and management of reproduction in Australian monotremes and marsupials
    Keeley, T ; Johnston, S ; Vogelnest, L ; Portas, T (CSIRO Publishing, 2019)
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    Proteomics and Deep Sequencing Comparison of Seasonally Active Venom Glands in the Platypus Reveals Novel Venom Peptides and Distinct Expression Profiles
    Wong, ESW ; Morgenstern, D ; Mofiz, E ; Gombert, S ; Morris, KM ; Temple-Smith, P ; Renfree, MB ; Whittington, CM ; King, GF ; Warren, WC ; Papenfuss, AT ; Belov, K (AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2012-11)
    The platypus is a venomous monotreme. Male platypuses possess a spur on their hind legs that is connected to glands in the pelvic region. They produce venom only during the breeding season, presumably to fight off conspecifics. We have taken advantage of this unique seasonal production of venom to compare the transcriptomes of in- and out-of-season venom glands, in conjunction with proteomic analysis, to identify previously undiscovered venom genes. Comparison of the venom glands revealed distinct gene expression profiles that are consistent with changes in venom gland morphology and venom volumes in and out of the breeding season. Venom proteins were identified through shot-gun sequenced venom proteomes of three animals using RNA-seq-derived transcripts for peptide-spectral matching. 5,157 genes were expressed in the venom glands, 1,821 genes were up-regulated in the in-season gland, and 10 proteins were identified in the venom. New classes of platypus-venom proteins identified included antimicrobials, amide oxidase, serpin protease inhibitor, proteins associated with the mammalian stress response pathway, cytokines, and other immune molecules. Five putative toxins have only been identified in platypus venom: growth differentiation factor 15, nucleobindin-2, CD55, a CXC-chemokine, and corticotropin-releasing factor-binding protein. These novel venom proteins have potential biomedical and therapeutic applications and provide insights into venom evolution.
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    N-Glycosylation Determines Ionic Permeability and Desensitization of the TRPV1 Capsaicin Receptor
    Veldhuis, NA ; Lew, MJ ; Abogadie, FC ; Poole, DP ; Jennings, EA ; Ivanusic, JJ ; Eilers, H ; Bunnett, NW ; McIntyre, P (AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2012-06-22)
    The balance of glycosylation and deglycosylation of ion channels can markedly influence their function and regulation. However, the functional importance of glycosylation of the TRPV1 receptor, a key sensor of pain-sensing nerves, is not well understood, and whether TRPV1 is glycosylated in neurons is unclear. We report that TRPV1 is N-glycosylated and that N-glycosylation is a major determinant of capsaicin-evoked desensitization and ionic permeability. Both N-glycosylated and unglycosylated TRPV1 was detected in extracts of peripheral sensory nerves by Western blotting. TRPV1 expressed in HEK-293 cells exhibited various degrees of glycosylation. A mutant of asparagine 604 (N604T) was not glycosylated but did not alter plasma membrane expression of TRPV1. Capsaicin-evoked increases in intracellular calcium ([Ca(2+)](i)) were sustained in wild-type TRPV1 HEK-293 cells but were rapidly desensitized in N604T TRPV1 cells. There was marked cell-to-cell variability in capsaicin responses and desensitization between individual cells expressing wild-type TRPV1 but highly uniform responses in cells expressing N604T TRPV1, consistent with variable levels of glycosylation of the wild-type channel. These differences were also apparent when wild-type or N604T TRPV1-GFP fusion proteins were expressed in neurons from trpv1(-/-) mice. Capsaicin evoked a marked, concentration-dependent increase in uptake of the large cationic dye YO-PRO-1 in cells expressing wild-type TRPV1, indicative of loss of ion selectivity, that was completely absent in cells expressing N604T TRPV1. Thus, TRPV1 is variably N-glycosylated and glycosylation is a key determinant of capsaicin regulation of TRPV1 desensitization and permeability. Our findings suggest that physiological or pathological alterations in TRPV1 glycosylation would affect TRPV1 function and pain transmission.
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    Conserved Glu-47 and Lys-50 residues are critical for UDP-N-acetylglucosamine/UMP antiport activity of the mouse Golgi-associated transporter Slc35a3
    Agustina Toscanini, M ; Belen Favarolo, M ; Gonzalez Flecha, FL ; Ebert, B ; Rautengarten, C ; Bredeston, LM (AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2019-06-28)
    Nucleotide sugar transporters (NSTs) regulate the flux of activated sugars from the cytosol into the lumen of the Golgi apparatus where glycosyltransferases use them for the modification of proteins, lipids, and proteoglycans. It has been well-established that NSTs are antiporters that exchange nucleotide sugars with the respective nucleoside monophosphate. Nevertheless, information about the molecular basis of ligand recognition and transport is scarce. Here, using topology predictors, cysteine-scanning mutagenesis, expression of GFP-tagged protein variants, and phenotypic complementation of the yeast strain Kl3, we identified residues involved in the activity of a mouse UDP-GlcNAc transporter, murine solute carrier family 35 member A3 (mSlc35a3). We specifically focused on the putative transmembrane helix 2 (TMH2) and observed that cells expressing E47C or K50C mSlc35a3 variants had lower levels of GlcNAc-containing glycoconjugates than WT cells, indicating impaired UDP-GlcNAc transport activity of these two variants. A conservative substitution analysis revealed that single or double substitutions of Glu-47 and Lys-50 do not restore GlcNAc glycoconjugates. Analysis of mSlc35a3 and its genetic variants reconstituted into proteoliposomes disclosed the following: (i) all variants act as UDP-GlcNAc/UMP antiporters; (ii) conservative substitutions (E47D, E47Q, K50R, or K50H) impair UDP-GlcNAc uptake; and (iii) substitutions of Glu-47 and Lys-50 dramatically alter kinetic parameters, consistent with a critical role of these two residues in mSlc35a3 function. A bioinformatics analysis revealed that an EXXK motif in TMH2 is highly conserved across SLC35 A subfamily members, and a 3D-homology model predicted that Glu-47 and Lys-50 are facing the central cavity of the protein.
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    Using decision support tools in emergency animal disease planning and response: Foot and mouth disease (CEBRA Project 1404D), Technical Report prepared for the Department of Agriculture and Water Resources
    Garner, G ; East, I ; Bradhurst, R ; Roche, S ; Rawdon, T ; Sanson, R ; Kompas, T ; Van Pham, H ; Stevenson, M (University of Melbourne, 2016)
    Modelling studies both in Australia and overseas have shown that vaccination can be very effective in reducing the size and duration of an FMD outbreak. Vaccination is most effective in reducing the duration and size of an outbreak when used early and is less effective the longer you delay. However, a decision to vaccinate early in the outbreak may result in using vaccination in situations where it is not actually required, with consequent implications for post-outbreak surveillance, the management of vaccinated animals and the ability to regain FMD-free status and access to markets. Overall, the choice of control measure to adopt in an FMD outbreak will thus depend on the variable and potentially conflicting objectives of the control program. As an important component of disease planning and preparedness for the department, the project will report on key information that could be used in an FMD outbreak to infer the potential scale of an outbreak and information to support disease management decision-making.
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    Incorporating economic components in Australia's FMD modelling capability and evaluating post-outbreak management to support return to trade (CEBRA project 1608D), Technical Report for the Department of Agriculture, Water and Environment
    Garner, G ; Bradhurst, R ; Death, C ; Dodd, A ; East, I ; Kompas, T (University of Melbourne, 2017)
    Following an outbreak of FMD, surveillance will be required to demonstrate that infection has been eradicated from the population and enable any remaining movement restrictions to be lifted within the country. Proof of freedom will also be needed to satisfy trading partners and regain access to international markets. Although vaccination is increasingly being recognised as an important tool to assist in containing and eradicating FMD outbreaks, it will make achieving recognition of free status more difficult—keeping vaccinated animals in the population will delay the period until FMD-free status is regained under the World Organisation for Animal Health (OIE) guidelines and add additional complications to the postoutbreak surveillance program. There is no agreed approach to post-outbreak management of vaccinated animals in AUSVETPLAN with the options being to: (1) allow vaccinated animals to remain in the population to live out their normal commercial lives (vaccinate-to-live); (2) remove all vaccinated animals from the population (vaccinateand- remove). Under option 2, vaccinated animals could be subject to (a) slaughter to waste i.e. remove and dispose of vaccinated animals; or (b) slaughter and salvage i.e. attempt to sell either raw or processed product from vaccinated animals. For (b) there may be some residual value of products that could offset some of the costs. The project will bring together epidemiological and economic expertise from the Department, the Australian National University, and CEBRA to formally explore and establish a science-based and cost effective approach to regaining free-status after an FMD outbreak as expeditiously as possible. The project will expand the Department’s modelling capability as well as providing insights into postoutbreak FMD management and contribute to Australia’s FMD preparedness.
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    National-level farm demographic data for preparedness of highly-infectious livestock disease epidemics. Review of data sources in New Zealand, approach to modelling populations and the effect of population uncertainty on disease modelling.
    van Andel, M ; Hollings, T ; Robinson, A ; Jewell, C ; Burgman, M ; Vink, D ; Sattler, K ; Masako, W ; Carpenter, T ; Bradhurst, R ; Garner, G (University of Melbourne, 2016)
    Isolation and strict biosecurity measures implemented by Australia and New Zealand have prevented the incursion of many organisms of biosecurity concern. The agricultural industry is a key part of the economy for both countries, and preventing the arrival of diseases of concern, including foot-and-mouth (FMD) disease, is crucial to maintaining access to international markets, reputation, and protecting the economy and industry. Early detection of, and a rapid, effective response to such diseases have a large impact on limiting the economic damage caused by epidemics. An acknowledged weakness of biosecurity preparedness and response to agricultural diseases in both countries is the lack of a single source of accurate, up-to-date farm livestock demographics information. This report reviews the use of animal counts for investigation of, preparedness for and response to exotic animal disease outbreaks, and analyses the available New Zealand datasets in depth. Gaps and weaknesses in the current data landscape are documented. The project objectives then focus on developing methodologies to estimate national-level farm demographic data and assess the use of modelled and inaccurate data in disease simulation models. There are nine key deliverables outlined in the report which were carried out over the two year duration of the project.
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    Cost-benefit analysis of the yellow crazy ant eradication program. Technical Report prepared for the Wet Tropics Management Authority
    Spring, D ; Kompas, T ; Bradhurst, R (Centre of Excellence for Biosecurity Risk Analysis, 2019)
    Yellow crazy ants (Anoplolepis gracilipes) (YCA) are one of the world’s 100 worst invasive species (Lowe et al. 2000). Previous assessments of YCA invasions have demonstrated that YCA can dramatically reduce native species richness in invaded areas, including in the Seychelles (Bos et al. 2008), Christmas Island (O'Dowd et al. 2003), and Hawaii (Plentovich et al. 2011). Native species losses include direct losses of competing invertebrate species and indirect losses resulting from ecological interdependencies, which can result in “ecological meltdown” in extreme cases such as Christmas Island (O'Dowd et al. 2003). YCA can also cause large losses to people living in infested areas through nuisance and health effects (Lach and Hoskin 2015) and can also adversely affect agricultural producers (Young et al. 2001) through reducing yields and/or increasing pesticide costs. YCA was first detected in Cairns and its southern suburbs in 2001, and an eradication program was initiated by the Department of Natural Resources and Mines (DNRM) and Biosecurity Queensland as part of a larger state-wide program. Later discoveries of YCA across the state, including in and around the WTWHA led to the state-wide eradication program being discontinued. An application was then made by WTMA to continue eradication efforts in and around the WTWHA. The program has been funded by the Australian Government and the Queensland Government in two overlapping projects, as described in the Executive Summary.
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    Widespread cis-regulatory convergence between the extinct Tasmanian tiger and gray wolf
    Feigin, CY ; Newton, AH ; Pask, AJ (COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT, 2019-10)
    The extinct marsupial Tasmanian tiger, or thylacine, and the eutherian gray wolf are among the most widely recognized examples of convergent evolution in mammals. Despite being distantly related, these large predators independently evolved extremely similar craniofacial morphologies, and evidence suggests that they filled similar ecological niches. Previous analyses revealed little evidence of adaptive convergence between their protein-coding genes. Thus, the genetic basis of their convergence is still unclear. Here, we identified candidate craniofacial cis-regulatory elements across vertebrates and compared their evolutionary rates in the thylacine and wolf, revealing abundant signatures of convergent positive selection. Craniofacial thylacine-wolf accelerated regions were enriched near genes involved in TGF beta (TGFB) and BMP signaling, both of which are key morphological signaling pathways with critical roles in establishing the identities and boundaries between craniofacial tissues. Similarly, enhancers of genes involved in craniofacial nerve development showed convergent selection and involvement in these pathways. Taken together, these results suggest that adaptation in cis-regulators of TGF beta and BMP signaling may provide a mechanism to explain the coevolution of developmentally and functionally integrated craniofacial structures in these species. We also found that despite major structural differences in marsupial and eutherian brains, accelerated regions in both species were common near genes with roles in brain development. Our findings support the hypothesis that, relative to protein-coding genes, positive selection on cis-regulatory elements is likely to be an essential driver of adaptive convergent evolution and may underpin thylacine-wolf phenotypic similarities.
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    Insect Antennal Morphology: The Evolution of Diverse Solutions to Odorant Perception
    Elgar, MA ; Zhang, D ; Wang, Q ; Wittwer, B ; Hieu, TP ; Johnson, TL ; Freelance, CB ; Coquilleau, M (Yale University, 2018-12-01)
    Chemical communication involves the production, transmission, and perception of odors. Most adult insects rely on chemical signals and cues to locate food resources, oviposition sites or reproductive partners and, consequently, numerous odors provide a vital source of information. Insects detect these odors with receptors mostly located on the antennae, and the diverse shapes and sizes of these antennae (and sensilla) are both astonishing and puzzling: what selective pressures are responsible for these different solutions to the same problem - to perceive signals and cues? This review describes the selection pressures derived from chemical communication that are responsible for shaping the diversity of insect antennal morphology. In particular, we highlight new technologies and techniques that offer exciting opportunities for addressing this surprisingly neglected and yet crucial component of chemical communication.