The Experts below are selected from a list of 252 Experts worldwide ranked by ideXlab platform
Robert J Beynon - One of the best experts on this subject based on the ideXlab platform.
-
molecular complexity of the Major Urinary protein system of the norway rat rattus norvegicus
Scientific Reports, 2019Co-Authors: Guadalupe Gomezbaena, Jane L Hurst, Stuart D Armstrong, Sarah A Roberts, Lynn Mclean, Josiah O Halstead, Mark Prescott, Jonathan M Mudge, Robert J BeynonAbstract:Major Urinary Proteins (MUP) are the Major component of the Urinary protein fraction in house mice (Mus spp.) and rats (Rattus spp.). The structure, polymorphism and functions of these lipocalins have been well described in the western European house mouse (Mus musculus domesticus), clarifying their role in semiochemical communication. The complexity of these roles in the mouse raises the question of similar functions in other rodents, including the Norway rat, Rattus norvegicus. Norway rats express MUPs in urine but information about specific MUP isoform sequences and functions is limited. In this study, we present a detailed molecular characterization of the MUP proteoforms expressed in the urine of two laboratory strains, Wistar Han and Brown Norway, and wild caught animals, using a combination of manual gene annotation, intact protein mass spectrometry and bottom-up mass spectrometry-based proteomic approaches. Cluster analysis shows the existence of only 10 predicted mup genes. Further, detailed sequencing of the Urinary MUP isoforms reveals a less complex pattern of primary sequence polymorphism in the rat than the mouse. However, unlike the mouse, rat MUPs exhibit added complexity in the form of post-translational modifications, including the phosphorylation of Ser4 in some isoforms, and exoproteolytic trimming of specific isoforms. Our results raise the possibility that Urinary MUPs may have different roles in rat chemical communication than those they play in the house mouse. Shotgun proteomics data are available via ProteomExchange with identifier PXD013986.
-
molecular complexity of the Major Urinary protein system of the norway rat rattus norvegicus
bioRxiv, 2018Co-Authors: Guadalupe Gomezbaena, Jane L Hurst, Stuart D Armstrong, Sarah A Roberts, Lynn Mclean, Josiah O Halstead, Mark Prescott, Jonathan M Mudge, Robert J BeynonAbstract:Major Urinary Proteins (MUP) are the Major component of the Urinary protein fraction in house mice (Mus spp.) and rats (Rattus spp.). The structure, polymorphism and functions of these lipocalins have been well described in the western European house mouse (Mus musculus domesticus), clarifying their role in semiochemical communication. The complexity of these roles in the mouse raises the question of similar functions in other rodents, including the Norway rat, Rattus norvegicus. Norway rats express MUPs in urine but information about specific MUP isoform sequences and functions is limited. In this study, we present a detailed molecular characterization of the MUP proteoforms expressed in the urine of two laboratory strains, Wistar Han and Brown Norway, and wild caught animals, using a combination of manual gene annotation, intact protein mass spectrometry and bottom-up mass spectrometry-based proteomic approaches. Detailed sequencing of the Proteins reveals a less complex pattern of primary sequence polymorphism than the mouse. However, unlike the mouse, rat MUPs exhibit added complexity in the form of post-translational modifications including phosphorylation and exoproteolytic trimming of specific isoforms. The possibility that Urinary MUPs may have different roles in rat chemical communication than those they play in the house mouse is also discussed.
-
individual odour signatures that mice learn are shaped by involatile Major Urinary Proteins mups
BMC Biology, 2018Co-Authors: Sarah A Roberts, Robert J Beynon, Amanda J Davidson, Mark C Prescott, Lynn Mclean, Jane L HurstAbstract:Reliable recognition of individuals requires phenotypic identity signatures that are both individually distinctive and appropriately stable over time. Individual-specific vocalisations or visual patterning are well documented among birds and some mammals, whilst odours play a key role in social recognition across many vertebrates and invertebrates. Less well understood, though, is whether individuals are recognised through variation in cues that arise incidentally from a wide variety of genetic and non-genetic differences between individuals, or whether animals evolve distinctive polymorphic signals to advertise identity reliably. As a bioassay to understand the derivation of individual-specific odour signatures, we use female attraction to the individual odours of male house mice (Mus musculus domesticus), learned on contact with a male’s scent marks. Learned volatile odour signatures are determined predominantly by individual differences in involatile Major Urinary protein (MUP) signatures, a specialised set of communication Proteins that mice secrete in their urine. Recognition of odour signatures in genetically distinct mice depended on differences in individual MUP genotype. Direct manipulation using recombinant MUPs confirmed predictable changes in volatile signature recognition according to the degree of matching between MUP profiles and the learned urine template. Both the relative amount of the male-specific MUP pheromone darcin, which induces odour learning, and other MUP isoforms influenced learned odour signatures. By contrast, odour recognition was not significantly influenced by individual Major histocompatibility complex genotype. MUP profiles shape volatile odour signatures through isoform-specific differences in binding and release of Urinary volatiles from scent deposits, such that volatile signatures were recognised from the Urinary protein fraction alone. Manipulation using recombinant MUPs led to quantitative changes in the release of known MUP ligands from scent deposits, with MUP-specific and volatile-specific effects. Despite assumptions that many genes contribute to odours that can be used to recognise individuals, mice have evolved a polymorphic combinatorial MUP signature that shapes distinctive volatile signatures in their scent. Such specific signals may be more prevalent within complex body odours than previously realised, contributing to the evolution of phenotypic diversity within species. However, differences in selection may also result in species-specific constraints on the ability to recognise individuals through complex body scents.
-
mass spectrometry for structural analysis and quantification of the Major Urinary Proteins of the house mouse
International Journal of Mass Spectrometry, 2015Co-Authors: Robert J Beynon, Jane L Hurst, Stuart D Armstrong, Amy J Claydon, Amanda J Davidson, Claire E Eyers, James I Langridge, Guadalupe Gomezbaena, Victoria M Harman, Victoria LeeAbstract:Abstract The Major Urinary Proteins (MUPs) of the house mouse, Mus musculus domesticus, are 18–19 kDa beta-barrel lipocalins that are involved in chemical communication between individuals. Many of them are excreted in urine where they play multiple roles, including coding of owner identity and transport, and slow release of bound volatile pheromones. One of them, darcin, is a pheromone in its own right and induces long-term memory for the identity and location of the scent mark owner. We have shown that mass spectrometric analysis of intact Proteins, and their ion mobility behaviour, is capable of dissecting subtle structural differences between the members of this class of Proteins. Moreover, mass spectrometric analysis of the intact Proteins can contribute towards molecular phenotyping of MUPs. However, whilst allowing relative quantification, the ionisation propensity or gas phase properties of the individual MUPs may compromise absolute quantification. To solve the challenge of absolute quantification of MUP expression, we have designed and constructed a QconCAT built from endopeptidase LysC peptides that is capable of quantifying MUPs found in laboratory animal strains and some MUPs from wild caught individuals.
-
limited variation in the Major Urinary Proteins of laboratory mice
Physiology & Behavior, 2009Co-Authors: Sarah A Cheetham, Robert J Beynon, Abigail L Smith, Stuart D Armstrong, Jane L HurstAbstract:Individual variation in a specialised set of scent communication Proteins, the Major Urinary Proteins (MUPs), provides a genetic identity signature that underlies individual and kin recognition, and the assessment of heterozygosity in wild house mice. Here we examine the extent to which MUP variation is retained among 30 classical strains of laboratory mice from three main lineages (Castle, C57, Swiss). Normal wild-type variation in Urinary MUP pattern appears to have been lost at an early stage in the derivation of the classical laboratory strains. All strains from the Castle and Swiss lineages shared the same "individual" MUP pattern, consistent with common ancestry from very few founders, while those from the C57 lineage shared a different pattern. Notably, individual variation in MUP pattern was no greater within the Swiss outbred ICR (CD-1) strain than typical for inbred strains. Total Urinary protein concentration varied considerably between even closely related substrains, together with minor variation in the relative amount of each MUP isoform expressed, although the functional significance of such quantitative variation in MUP expression has yet to be established. Expression was 2-8 fold higher among males, while a MUP expressed by most male but not female wild mice was expressed by C57 males but variably among Castle and Swiss males and occasionally by females in some strains. The lack of normal variation in MUP patterns within and between strains has important implications for the use of laboratory mice in behavioural or neurophysiological research investigating social recognition or mate choice.
Jane L Hurst - One of the best experts on this subject based on the ideXlab platform.
-
molecular complexity of the Major Urinary protein system of the norway rat rattus norvegicus
Scientific Reports, 2019Co-Authors: Guadalupe Gomezbaena, Jane L Hurst, Stuart D Armstrong, Sarah A Roberts, Lynn Mclean, Josiah O Halstead, Mark Prescott, Jonathan M Mudge, Robert J BeynonAbstract:Major Urinary Proteins (MUP) are the Major component of the Urinary protein fraction in house mice (Mus spp.) and rats (Rattus spp.). The structure, polymorphism and functions of these lipocalins have been well described in the western European house mouse (Mus musculus domesticus), clarifying their role in semiochemical communication. The complexity of these roles in the mouse raises the question of similar functions in other rodents, including the Norway rat, Rattus norvegicus. Norway rats express MUPs in urine but information about specific MUP isoform sequences and functions is limited. In this study, we present a detailed molecular characterization of the MUP proteoforms expressed in the urine of two laboratory strains, Wistar Han and Brown Norway, and wild caught animals, using a combination of manual gene annotation, intact protein mass spectrometry and bottom-up mass spectrometry-based proteomic approaches. Cluster analysis shows the existence of only 10 predicted mup genes. Further, detailed sequencing of the Urinary MUP isoforms reveals a less complex pattern of primary sequence polymorphism in the rat than the mouse. However, unlike the mouse, rat MUPs exhibit added complexity in the form of post-translational modifications, including the phosphorylation of Ser4 in some isoforms, and exoproteolytic trimming of specific isoforms. Our results raise the possibility that Urinary MUPs may have different roles in rat chemical communication than those they play in the house mouse. Shotgun proteomics data are available via ProteomExchange with identifier PXD013986.
-
molecular complexity of the Major Urinary protein system of the norway rat rattus norvegicus
bioRxiv, 2018Co-Authors: Guadalupe Gomezbaena, Jane L Hurst, Stuart D Armstrong, Sarah A Roberts, Lynn Mclean, Josiah O Halstead, Mark Prescott, Jonathan M Mudge, Robert J BeynonAbstract:Major Urinary Proteins (MUP) are the Major component of the Urinary protein fraction in house mice (Mus spp.) and rats (Rattus spp.). The structure, polymorphism and functions of these lipocalins have been well described in the western European house mouse (Mus musculus domesticus), clarifying their role in semiochemical communication. The complexity of these roles in the mouse raises the question of similar functions in other rodents, including the Norway rat, Rattus norvegicus. Norway rats express MUPs in urine but information about specific MUP isoform sequences and functions is limited. In this study, we present a detailed molecular characterization of the MUP proteoforms expressed in the urine of two laboratory strains, Wistar Han and Brown Norway, and wild caught animals, using a combination of manual gene annotation, intact protein mass spectrometry and bottom-up mass spectrometry-based proteomic approaches. Detailed sequencing of the Proteins reveals a less complex pattern of primary sequence polymorphism than the mouse. However, unlike the mouse, rat MUPs exhibit added complexity in the form of post-translational modifications including phosphorylation and exoproteolytic trimming of specific isoforms. The possibility that Urinary MUPs may have different roles in rat chemical communication than those they play in the house mouse is also discussed.
-
individual odour signatures that mice learn are shaped by involatile Major Urinary Proteins mups
BMC Biology, 2018Co-Authors: Sarah A Roberts, Robert J Beynon, Amanda J Davidson, Mark C Prescott, Lynn Mclean, Jane L HurstAbstract:Reliable recognition of individuals requires phenotypic identity signatures that are both individually distinctive and appropriately stable over time. Individual-specific vocalisations or visual patterning are well documented among birds and some mammals, whilst odours play a key role in social recognition across many vertebrates and invertebrates. Less well understood, though, is whether individuals are recognised through variation in cues that arise incidentally from a wide variety of genetic and non-genetic differences between individuals, or whether animals evolve distinctive polymorphic signals to advertise identity reliably. As a bioassay to understand the derivation of individual-specific odour signatures, we use female attraction to the individual odours of male house mice (Mus musculus domesticus), learned on contact with a male’s scent marks. Learned volatile odour signatures are determined predominantly by individual differences in involatile Major Urinary protein (MUP) signatures, a specialised set of communication Proteins that mice secrete in their urine. Recognition of odour signatures in genetically distinct mice depended on differences in individual MUP genotype. Direct manipulation using recombinant MUPs confirmed predictable changes in volatile signature recognition according to the degree of matching between MUP profiles and the learned urine template. Both the relative amount of the male-specific MUP pheromone darcin, which induces odour learning, and other MUP isoforms influenced learned odour signatures. By contrast, odour recognition was not significantly influenced by individual Major histocompatibility complex genotype. MUP profiles shape volatile odour signatures through isoform-specific differences in binding and release of Urinary volatiles from scent deposits, such that volatile signatures were recognised from the Urinary protein fraction alone. Manipulation using recombinant MUPs led to quantitative changes in the release of known MUP ligands from scent deposits, with MUP-specific and volatile-specific effects. Despite assumptions that many genes contribute to odours that can be used to recognise individuals, mice have evolved a polymorphic combinatorial MUP signature that shapes distinctive volatile signatures in their scent. Such specific signals may be more prevalent within complex body odours than previously realised, contributing to the evolution of phenotypic diversity within species. However, differences in selection may also result in species-specific constraints on the ability to recognise individuals through complex body scents.
-
mass spectrometry for structural analysis and quantification of the Major Urinary Proteins of the house mouse
International Journal of Mass Spectrometry, 2015Co-Authors: Robert J Beynon, Jane L Hurst, Stuart D Armstrong, Amy J Claydon, Amanda J Davidson, Claire E Eyers, James I Langridge, Guadalupe Gomezbaena, Victoria M Harman, Victoria LeeAbstract:Abstract The Major Urinary Proteins (MUPs) of the house mouse, Mus musculus domesticus, are 18–19 kDa beta-barrel lipocalins that are involved in chemical communication between individuals. Many of them are excreted in urine where they play multiple roles, including coding of owner identity and transport, and slow release of bound volatile pheromones. One of them, darcin, is a pheromone in its own right and induces long-term memory for the identity and location of the scent mark owner. We have shown that mass spectrometric analysis of intact Proteins, and their ion mobility behaviour, is capable of dissecting subtle structural differences between the members of this class of Proteins. Moreover, mass spectrometric analysis of the intact Proteins can contribute towards molecular phenotyping of MUPs. However, whilst allowing relative quantification, the ionisation propensity or gas phase properties of the individual MUPs may compromise absolute quantification. To solve the challenge of absolute quantification of MUP expression, we have designed and constructed a QconCAT built from endopeptidase LysC peptides that is capable of quantifying MUPs found in laboratory animal strains and some MUPs from wild caught individuals.
-
the effects of graded levels of calorie restriction ii impact of short term calorie and protein restriction on circulating hormone levels glucose homeostasis and oxidative stress in male c57bl 6 mice
Oncotarget, 2015Co-Authors: Sharon E Mitchell, Jane L Hurst, Camille Delville, Penelope Konstantopedos, Davina Derous, Cara L Green, Luonan Chen, Jackie J D Han, Ying Wang, Daniel E L PromislowAbstract:Limiting food intake attenuates many of the deleterious effects of aging, impacting upon healthspan and leading to an increased lifespan. Whether it is the overall restriction of calories (calorie restriction: CR) or the incidental reduction in macronutrients such as protein (protein restriction: PR) that mediate these effects is unclear. The impact of 3 month CR or PR, (10 to 40%), on C57BL/6 mice was compared to controls fed ad libitum. Reductions in circulating leptin, tumor necrosis factor-α and insulin-like growth factor-1 (IGF-1) were relative to the level of CR and individually associated with morphological changes but remained unchanged following PR. Glucose tolerance and insulin sensitivity were improved following CR but not affected by PR. There was no indication that CR had an effect on oxidative damage, however CR lowered antioxidant activity. No biomarkers of oxidative stress were altered by PR. CR significantly reduced levels of Major Urinary Proteins suggesting lowered investment in reproduction. Results here support the idea that reduced adipokine levels, improved insulin/IGF-1 signaling and reduced reproductive investment play important roles in the beneficial effects of CR while, in the short-term, attenuation of oxidative damage is not applicable. None of the positive effects were replicated with PR.
Pavel Stopka - One of the best experts on this subject based on the ideXlab platform.
-
transcriptomic and proteomic profiling revealed high proportions of odorant binding and antimicrobial defense Proteins in olfactory tissues of the house mouse
Frontiers in Genetics, 2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, whilst the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g. Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis.
-
Data_Sheet_4.XLSX
2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, while the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g., Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis.
-
Transcriptomic and Proteomic Profiling Revealed High Proportions of Odorant Binding and Antimicrobial Defense Proteins in Olfactory Tissues of the House Mouse
Frontiers Media S.A., 2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, while the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g., Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis
-
On the tear proteome of the house mouse (Mus musculus musculus) in relation to chemical signalling
PeerJ Inc., 2017Co-Authors: Romana Stopkova, Barbora Kuntova, Petr Klempt, Pavel StopkaAbstract:Mammalian tears are produced by lacrimal glands to protect eyes and may function in chemical communication and immunity. Recent studies on the house mouse chemical signalling revealed that Major Urinary Proteins (MUPs) are not individually unique in Mus musculus musculus. This fact stimulated us to look for other sexually dimorphic Proteins that may—in combination with MUPs—contribute to a pool of chemical signals in tears. MUPs and other lipocalins including odorant binding Proteins (OBPs) have the capacity to selectively transport volatile organic compounds (VOCs) in their eight-stranded beta barrel, thus we have generated the tear proteome of the house mouse to detect a wider pool of Proteins that may be involved in chemical signalling. We have detected significant male-biased (7.8%) and female-biased (7%) Proteins in tears. Those Proteins that showed the most elevated sexual dimorphisms were highly expressed and belong to MUP, OBP, ESP (i.e., exocrine gland-secreted peptides), and SCGB/ABP (i.e., secretoglobin) families. Thus, tears may have the potential to elicit sex-specific signals in combination by different Proteins. Some tear lipocalins are not sexually dimorphic—with MUP20/darcin and OBP6 being good examples—and because all Proteins may flow with tears through nasolacrimal ducts to nasal and oral cavities we suggest that their roles are wider than originally thought. Also, we have also detected several sexually dimorphic bactericidal Proteins, thus further supporting an idea that males and females may have adopted alternative strategies in controlling microbiota thus yielding different VOC profiles
-
Searching for Major Urinary Proteins (MUPs) as Chemosignals in Urine of Subterranean Rodents
Journal of Chemical Ecology, 2011Co-Authors: Petra Hagemeyer, Katerina Janotova, Petr L. Jedelsky, Sabine Begall, Josephine Todrank, Hynek Burda, Giora Heth, Pavel StopkaAbstract:Chemosensory information mediates behavior in many rodent genera. Major Urinary Proteins (MUPs) facilitate chemical communication in some species of mice. We sought to demonstrate the importance of MUPs in chemosignaling across a range of rodent genera that live in different habitats and social structures. We analyzed urine from three subterranean rodent genera from different continents, and with diverse social systems: eusocial Zambian mole-rats (Fukomys), solitary Israeli blind mole rats (Spalax), and social Chilean coruros (Spalacopus). 2D gel electrophoresis revealed low levels of protein, with sequences similar to aphrodisin, in Fukomys mole-rat urine, but no MUPs in urine of any of the studied species. Previous research demonstrated that subjects from the tested genera responded differentially to odors indicating transmission of individuality, family/colony or population, species, and reproductive state in secretions and excretions of conspecifics. This extends, to subterranean rodents, the evidence that rodent species can successfully transmit and receive chemosignals without the necessity of MUPs.
Duncan H L Robertson - One of the best experts on this subject based on the ideXlab platform.
-
dynamic instability of the Major Urinary protein gene family revealed by genomic and phenotypic comparisons between c57 and 129 strain mice
Genome Biology, 2008Co-Authors: Jonathan M Mudge, Robert J Beynon, Jane L Hurst, Duncan H L Robertson, Stuart D Armstrong, Karen Mclaren, Christine Nicholson, Laurens G Wilming, Jennifer HarrowAbstract:Background: The Major Urinary Proteins (MUPs) of Mus musculus domesticus are deposited in urine in large quantities, where they bind and release pheromones and also provide an individual 'recognition signal' via their phenotypic polymorphism. Whilst important information about MUP functionality has been gained in recent years, the gene cluster is poorly studied in terms of structure, genic polymorphism and evolution. Results: We combine targeted sequencing, manual genome annotation and phylogenetic analysis to compare the Mup clusters of C57BL/6J and 129 strains of mice. We describe organizational heterogeneity within both clusters: a central array of cassettes containing Mup genes highly similar at the protein level, flanked by regions containing Mup genes displaying significantly elevated divergence. Observed genomic rearrangements in all regions have likely been mediated by endogenous retroviral elements. Mup loci with coding sequences that differ between the strains are identified - including a gene/pseudogene pair - suggesting that these inbred lineages exhibit variation that exists in wild populations. We have characterized the distinct MUP profiles in the urine of both strains by mass spectrometry. The total MUP phenotype data is reconciled with our genomic sequence data, matching all Proteins identified in urine to annotated genes. Conclusion: Our observations indicate that the MUP phenotypic polymorphism observed in wild populations results from a combination of Mup gene turnover coupled with currently unidentified mechanisms regulating gene expression patterns. We propose that the structural heterogeneity described within the cluster reflects functional divergence within the Mup gene family.
-
Characterization and comparison of Major Urinary Proteins from the house mouse, Mus musculus domesticus, and the aboriginal mouse, Mus macedonicus.
Journal of chemical ecology, 2007Co-Authors: Duncan H L Robertson, Jane L Hurst, Jeremy B. Searle, İslam Gündüz, Robert J BeynonAbstract:Urine from the house mouse, Mus musculus domesticus, contains a high concentration of Major Urinary Proteins (MUPs), which convey olfactory information between conspecifics. In wild populations, each individual expresses a different pattern of around 8 to 14 electrophoretically separable MUP isoforms. To examine whether other Mus species express MUPs and exhibit a similar level of individual heterogeneity, we characterized Urinary Proteins in urine samples from an aboriginal species, Mus macedonicus, captured from different sites in Turkey. Anion exchange chromatography and electrospray ionization mass spectrometry demonstrated that M. macedonicus urine contained a single Major peak of mass 18,742 Da, and in contrast to M. m. domesticus, all individuals were the same. The M. macedonicus masses were not predicted from any known MUP gene sequence. Endoproteinase Lys-C (Lys-C) digestion of the purified M. macedonicus Urinary protein followed by matrix assisted laser desorption time of flight (MALDI-TOF) mass spectrometry demonstrated that it shared considerable, but not complete, sequence homogeneity with M. m. domesticus MUPs. Three M. macedonicus Lys-C peptides differed in mass from their M. m. domesticus counterparts. These three peptides were further characterized by tandem mass spectrometry. The complete sequences of two were determined, and in conjunction with methyl esterification, the amino acid composition of the third was inferred, and the sequence narrowed down to three permutations. The complete M. macedonicus sequence contained a maximum of seven amino acid substitutions, discernible by tandem mass spectrometry, relative to a reference M. m. domesticus sequence. Six of these were on the surface of the molecule. Molecular modeling of the M. macedonicus sequence demonstrated that the amino acid substitutions had little effect on the tertiary structure. The differences in the level of heterogeneity between the two species are discussed in relation to their environment and behavior. In addition, the differences in protein structure allow speculation into molecular mechanisms of MUP function.
-
structural and functional differences in isoforms of mouse Major Urinary Proteins a male specific protein that preferentially binds a male pheromone
Biochemical Journal, 2005Co-Authors: Stuart D Armstrong, Duncan H L Robertson, Jane L Hurst, Sarah A Cheetham, Robert J BeynonAbstract:The MUPs (Major Urinary Proteins) of the house mouse, Mus domesticus, are lipocalins that bind and slowly release male-specific pheromones in deposited scent marks. However, females also express these Proteins, consistent with a second role in encoding individual signatures in scent marks. We have purified and characterized an atypical MUP from the urine of male C57BL/6J inbred mice, which is responsible for the binding of most of the male pheromone, 2-sec-butyl-4,5-dihydrothiazole, and which is also responsible for the slow release of this pheromone from scent marks. This protein is absent from the urine of female mice of the same strain. The protein has been characterized by MS, leading to unequivocal identification as a previously uncharacterized gene product, providing compelling evidence for the expression of this gene in liver and manifestation in urine. These properties contrast strongly with those of the other MUPs in the same urine sample, and suggest that the requirement to manifest a male-specific pheromone has been met by evolution of a cognate protein specifically adapted to the binding and release of this ligand. This atypical MUP is also present in a random sample of wild-caught male mice, confirming that this protein is not specific to the inbred mouse strain but is present in natural populations also.
-
polymorphism in Major Urinary Proteins molecular heterogeneity in a wild mouse population
Journal of Chemical Ecology, 2002Co-Authors: Robert J Beynon, Caroline E Payne, Richard E Humphries, Christina Veggerby, Simon J Gaskell, Duncan H L Robertson, Jane L HurstAbstract:Major Urinary Proteins (MUPs) are present in high levels in the urine of mice, and the specific profile of MUPs varies considerably among wild-caught individuals. We have conducted a detailed study of the polymorphic variation within a geographically constrained island population, analyzing the MUP heterogeneity by isoelectric focusing and analytical ion exchange chromatography. Several MUPs were purified in sufficient quantities for analysis by electrospray ionization mass spectrometry and MALDI-TOF mass spectrometry of endopeptidase Lys-C peptide maps. The results of such analyses permitted the identification of three new MUP allelic variants. In each of these Proteins, the sites of variation were located to a restricted segment of the polypeptide chain, projecting to a patch on the surface of the protein, and connected to the central lipocalin calyx through the polypeptide backbone. The restriction of the polymorphic variation to one segment of the polypeptide may be of functional significance, either in the modulation of ligand release or in communication of individuality signals within Urinary scent marks.
-
individual recognition in mice mediated by Major Urinary Proteins
Nature, 2001Co-Authors: Jane L Hurst, Caroline E Payne, Richard E Humphries, Duncan H L Robertson, Charlotte M Nevison, Amr Darwish Marie, Andrea Cavaggioni, Robert J BeynonAbstract:The ability to recognize individuals is essential to many aspects of social behaviour, such as the maintenance of stable social groups, parent-offspring or mate recognition, inbreeding avoidance and the modulation of competitive relationships. Odours are a primary mediator of individuality signals among many mammals. One source of odour complexity in rodents, and possibly in humans, resides in the highly polymorphic Major histocompatibility complex (MHC). The olfactory acuity of mice and rats allows them to distinguish between the Urinary odours of congenic strains differing only in single genes within the MHC, although the chemical mediators or odorants are unknown. However, rodent urine also contains a class of Proteins, termed Major Urinary Proteins (MUPs), that bind and release small volatile pheromones. We have shown that the combinatorial diversity of expression of MUPs among wild mice might be as great as for MHC, and at protein concentrations a million times higher. Here we show in wild house mice (Mus domesticus) that Urinary MUPs play an important role in the individual recognition mechanism.
Romana Stopkova - One of the best experts on this subject based on the ideXlab platform.
-
transcriptomic and proteomic profiling revealed high proportions of odorant binding and antimicrobial defense Proteins in olfactory tissues of the house mouse
Frontiers in Genetics, 2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, whilst the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g. Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis.
-
Data_Sheet_4.XLSX
2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, while the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g., Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis.
-
Transcriptomic and Proteomic Profiling Revealed High Proportions of Odorant Binding and Antimicrobial Defense Proteins in Olfactory Tissues of the House Mouse
Frontiers Media S.A., 2018Co-Authors: Barbora Kuntova, Romana Stopkova, Pavel StopkaAbstract:Mammalian olfaction depends on chemosensory neurons of the main olfactory epithelia (MOE), and/or of the accessory olfactory epithelia in the vomeronasal organ (VNO). Thus, we have generated the VNO and MOE transcriptomes and the nasal cavity proteome of the house mouse, Mus musculus musculus. Both transcriptomes had low levels of sexual dimorphisms, while the soluble proteome of the nasal cavity revealed high levels of sexual dimorphism similar to that previously reported in tears and saliva. Due to low levels of sexual dimorphism in the olfactory receptors in MOE and VNO, the sex-specific sensing seems less likely to be dependent on receptor repertoires. However, olfaction may also depend on a continuous removal of background compounds from the sites of detection. Odorant binding Proteins (OBPs) are thought to be involved in this process and in our study Obp transcripts were most expressed along other lipocalins (e.g., Lcn13, Lcn14) and antimicrobial Proteins. At the level of proteome, OBPs were highly abundant with only few being sexually dimorphic. We have, however, detected the Major Urinary Proteins MUP4 and MUP5 in males and females and the male-biased central/group-B MUPs that were thought to be abundant mainly in the urine. The exocrine gland-secreted peptides ESP1 and ESP22 were male-biased but not male-specific in the nose. For the first time, we demonstrate that the expression of nasal lipocalins correlates with antimicrobial Proteins thus suggesting that their individual variation may be linked to evolvable mechanisms that regulate natural microbiota and pathogens that regularly enter the body along the ‘eyes-nose-oral cavity’ axis
-
On the tear proteome of the house mouse (Mus musculus musculus) in relation to chemical signalling
PeerJ Inc., 2017Co-Authors: Romana Stopkova, Barbora Kuntova, Petr Klempt, Pavel StopkaAbstract:Mammalian tears are produced by lacrimal glands to protect eyes and may function in chemical communication and immunity. Recent studies on the house mouse chemical signalling revealed that Major Urinary Proteins (MUPs) are not individually unique in Mus musculus musculus. This fact stimulated us to look for other sexually dimorphic Proteins that may—in combination with MUPs—contribute to a pool of chemical signals in tears. MUPs and other lipocalins including odorant binding Proteins (OBPs) have the capacity to selectively transport volatile organic compounds (VOCs) in their eight-stranded beta barrel, thus we have generated the tear proteome of the house mouse to detect a wider pool of Proteins that may be involved in chemical signalling. We have detected significant male-biased (7.8%) and female-biased (7%) Proteins in tears. Those Proteins that showed the most elevated sexual dimorphisms were highly expressed and belong to MUP, OBP, ESP (i.e., exocrine gland-secreted peptides), and SCGB/ABP (i.e., secretoglobin) families. Thus, tears may have the potential to elicit sex-specific signals in combination by different Proteins. Some tear lipocalins are not sexually dimorphic—with MUP20/darcin and OBP6 being good examples—and because all Proteins may flow with tears through nasolacrimal ducts to nasal and oral cavities we suggest that their roles are wider than originally thought. Also, we have also detected several sexually dimorphic bactericidal Proteins, thus further supporting an idea that males and females may have adopted alternative strategies in controlling microbiota thus yielding different VOC profiles
-
novel obp genes similar to hamster aphrodisin in the bank vole myodes glareolus
BMC Genomics, 2010Co-Authors: Romana Stopkova, Stěpan Ryba, Martin Sandera, Ondřej Šedo, Zbyněk Zdrahal, Pavel StopkaAbstract:Chemical communication in mammals involves globular lipocalins that protect and transport pheromones during their passage out of the body. Efficient communication via this protein - pheromone complex is essential for triggering multiple responses including aggression, mate choice, copulatory behaviour, and onset and synchronization of oestrus. The roles of lipocalins in communication were studied in many organisms and especially in mice (i.e. Mus musculus domesticus) which excrete Major Urinary Proteins (Mup) in excessive amounts in saliva and urine. Other mammals, however, often lack the genes for Mups or their expression is very low. Therefore, we aimed at characterization of candidate lipocalins in Myodes glareolus which are potentially linked to chemical communication. One of them is Aphrodisin which is a unique lipocalin that was previously described from hamster vaginal discharge and is known to carry pheromones stimulating copulatory behaviour in males. Here we show that Aphrodisin-like Proteins exist in other species, belong to a group of Odorant Binding Proteins (Obp), and contrary to the expression of Aphrodisin only in hamster genital tract and parotid glands of females, we have detected these transcripts in both sexes of M. glareolus with the expression confirmed in various tissues including prostate, prepucial and salivary glands, liver and uterus. On the level of mRNA, we have detected three different gene variants. To assess their relevance for chemical communication we investigated the occurrence of particular Proteins in saliva, urine and vaginal discharge. On the protein level we confirmed the presence of Obp2 and Obp3 in both saliva and urine. Appropriate bands in the range of 17-20 kDa from vaginal discharge were, however, beyond the MS detection limits. Our results demonstrate that three novel Obps (Obp1, Obp2, and Obp3) are predominant lipocalins in Myodes urine and saliva. On the protein level we have detected further variants and thus we assume that similarly as Major Urinary Proteins in mice, these Proteins may be important in chemical communication in this Cricetid rodent.
