Project description:Communication between individuals via molecules, termed chemosignaling, is widespread among animal and plant species. However, we lack knowledge on the specific functions of the substances involved for most of such systems. The femoral gland is an organ secreting a waxy substance, which is involved in chemical communication in lizards. While the lipids and volatile substances secreted by the femoral glands have been investigated in several biochemical studies, the protein composition and functions of secretions are completely unknown. Applying a proteomic approach, we provide the first attempt to comprehensively characterize the protein composition of femoral gland secretions from the Galápagos marine iguana. Using samples from several organs, the marine iguana proteome was assembled by next generation sequencing and mass spectrometry, resulting in 7,513 proteins; 4305 proteins were present in the femoral gland, including keratins, small serum proteins and fatty acid-binding proteins. Surprisingly, no proteins with obvious roles in partner recognition or inter-species communication could be identified. However, we found several proteins with direct associations to the innate immune system, such as lysozyme C, antileukoproteinase (ALP), pulmonary surfactant protein (SFTPD) and galectin (LGALS1), suggesting the femoral glands function as an important barrier to infection. The identification of previously unrecognized effective anti-microbial peptides from the femoral gland show similar activities against E.coli and B. subtilis compared to oncocin. This proteomics dataset is a valuable resource for future functional protein analysis and demonstrates that femoral glands secretions covers also functions of the innate immune system.
Project description:Background: Anopheles culicifacies is a rural vector of malaria in tropical and sub tropical South East Asian region. The salivary gland of the mosquito is the target for sporozoite interaction, blood feeding behavior, haemostasis and vector-parasite interactions. Malaria parasite matures inside the salivary gland, gain competence and transmitted to the host along with the saliva during biting. The importance of the proteins expressed in salivary gland is the first step in understanding the physiology of blood feeding and may provide insights into vector- parasite interactions. Since, no genomic or transcriptomics information is available of Anopheles culicifacies, therefore locally expressed functional proteins in salivary glands are of much importance. . Method: In this study, 1DE protein and in solution digestion was combined with tandem mass spectrometry (nano LC-MS/MS) and computational bioinformatics for data mining was employed to study the proteome profile of salivary glands of sugar fed An. culicifacies mosquito species. Functional annotation of all the identified proteins was carried out using gene ontology tools, CELLO and SMART analysis software. Results: Total 102 proteins were identified and analysed by SEQUEST algorithm against mosquito protein database from Uniprot/NCBI. Out of which 81 proteins were identified using gel free approach and 21 proteins using in-gel approach and 15 were common among these two approaches. All the identified proteins were categorized in to 23 groups of biological processes using GO tool. 7 proteins were depicted to be secretary in nature by investigating the signal peptide present. Potential proteins with unknown function were predicted by analyzing their functional association with other characterized proteins by STRING algorithm and were categorized in cell adhesion, cytoskeleton and membrane trafficking networks. Conclusion: Our study elucidates the first proteomic dataset of An. culicifacies salivary gland proteins. Functional annotation of salivary proteins and complementary gene ontology assignments in An. culicifacies species may contribute towards understanding the complex physiology of the tissues in this species. This proteome baseline data may facilitate the discernment of salivary glands and parasite correlation during blood feeding. Furthermore, this mass spectrometry based proteomic data may also provide insights into the elucidation of role of differential functional proteins present in refractory An. culicifacies mosquito and may be useful for development of effective malaria control strategies.
Project description:Cross-species hybridization analysis of mammary glands during pregnancy and lactation. Results provide insight into putative conserved molecular mechanisms regulating mammary gland development. This study was performed to identify orthologous transcripts that are differentially co-expressed in the mammary gland at 2 stages of development (pregnancy and lactation) in wild type Sprague-Dawley rats.
Project description:To expand the knowledge on the porcine salivary proteome, secretions from the three major salivary glands were collected from anaesthetised piglets. Pilocarpine and isoproterenol were simultaneously injected intraperitoneally to increase the volume and protein concentration of the saliva, respectively. The protein composition and relative protein-specific abundance of saliva secreted by the parotid gland and by the mandibular and monostomatic sublingual gland, were determined using iTRAQ. When combining two detection methods, MALDI-TOF/TOF MS and Q-Exactive orbitrap MS/MS, a total of 122 porcine salivary proteins and 6 mammalian proteins with a predicted porcine homolog were identified. Only a quantitative and not a qualitative difference was observed between both ductal secretions. The 128 proteins were detected in both secretions, however, at different levels. Twenty-four proteins (20 porcine, 4 mammalian with a predicted porcine homolog) were predominantly secreted by the parotid gland, such as carbonic anhydrase VI and alpha-amylase. Twenty-nine proteins (all porcine) were predominantly secreted by the mandibular and sublingual glands, for example salivary lipocalin and submaxillary apomucin protein.
Project description:In this project, the total proteomics of Rhipicephalus haemaphysaloides salivary gland in different periods (Fed3, Fed5, E0h and E72h) were analyzed. Quantitative Proteomic Analysis by Tandem Mass Tag (TMT) technology was used.
Project description:Chemical signaling through pheromones is an ancient and widespread mode of communication. Turtles and tortoises (chelonians) secrete pheromones via mental (chin) glands and have superior olfactory capacities rendering them a promising group to study the evolution and function of chemical communication in vertebrates. Here, we use state-of-the art proteomics and lipidomics techniques to identify and explore the possible functions of proteins and lipids secreted by mental glands in turtles. We examined four turtle species all from the family Geoemydidae, to understand among-species as well as sexual variation in the composition of mental gland secretions. Differential expression of a relatively small number (ca. 65) of proteins explained a large portion of the proteome variation across species, highlighting the existence of specific signals evolving even in closely related species. Lipidomic analysis revealed high inter-individual variation but species differences could be attributed to five differing lipid classes. The lipids found in mental glands could have a dietary origin or be related to individual condition but may nonetheless be used in communication. We also examined sex-specific differences in the proteome of a single species (Mauremys leprosa) and found that males expressed a much larger array of proteins than females. Our findings establish a group of candidate proteins potentially involved in chemical signaling freshwater geoemydid turtles. Alternatively, differently expressed proteins in mental glands could have an indirect link to chemical communication, being involved in pheromone transport and/or antioxidant protection.
Project description:Proteomic analysis of cytokines in unstimulated oropharyngeal secretions. Epstein-barr virus (EBV) is a type 1 carcinogen which causes many cancers in humans. Here we explored the cytokine involvement of the EBV replication process in the oropharynx. Cytokine interactomic profiles were geneerated to understand the involved signalling pathways in HIV infected group and the healthy group. Proteome profilers were used to understand the major cytokine expression levels that are related to infection and immune regulation.
Project description:Cross-species hybridization analysis of mammary glands during pregnancy and lactation. Results provide insight into putative conserved molecular mechanisms regulating mammary gland development. This study was performed to identify orthologous transcripts that are differentially co-expressed in the mammary gland at 2 stages of development (pregnancy and lactation) in wild type Sprague-Dawley rats. Key points are examined in a time series of Sprague Dawley rat mammary gland development, secretory activation and lactation. Triplicate rat (three biological replicates) at each time point were used for statistical power totalling 12 individual arrays in this study. Rats were as staged pregnant day 1 the day that post coital plug was observed, and similarly, lactation day 1 was the first day after birth. Whole mammary glands No. 4 (inguinal) were obtained from female rats at stages of development: virgin (adulthood, 14 wks of age), Pregnant (5 and 14 days of pregnancy) and Lactating (day 1 and 12 postpartum). The two-color (Cy5/Cy3) microarray experiment was designed to hybridize samples from each group against a common reference, a pool of RNA from mammary gland of three parous or virgin female rats.