Progress 07/01/09 to 06/30/14
Outputs
Target Audience: Participants: Two postdoctoral researcher and two graduate students were trained. * Ladislav Simo (Postdoctoral researcher) * Jurak Koci (Postdoctoral researcher) * Donghun Andy Kim (Ph.D. student) * Joshua Urban (Ph.D. student) Target Audiences: We have made total 23 invited presentations (1 to 23) and 26 contributed presentations (24 to 49) as direct activities of this proposal and 34 additional presentations (50 to 83) related to the proposed activity in local, national, and international conferences. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Outputs: The proposed activities were with three aims listed below. 1. Identifying candidate neuropeptides relevant to salivary functions. 2. Identifying the G protein-coupled receptors for dopamine, PGE2, MIP, and SIFamide. 3. Investigation of the functions of signaling systems in control of salivary secretion. We have completed all three aims except the study of PGE2 receptor study. Unfortunately, however, RNA interference (RNAi) for functional studies found that the RNAi in tick species are tissue specific or gene specific, causing difficulties in achieving the proposed Aim 3. We have been developing a technique for in vitro microinjections of bioactive compounds in the acini to overcome this problem in the functional study. Outcome/Impacts: We have completed a part of Specific Aim 1 and 2 and a part of Specific Aim 3. These accomplishments were presented in international, national, and local conferences and published in peer-reviewed journals as they are listed.
Publications
- Type:
Other
Status:
Published
Year Published:
2014
Citation:
1. Kim, D., `imo, L and Y. Park (2014) Orchestration of salivary secretion mediated by two different dopamine receptors in the backlogged tick Ixodes scapularis. J. Exp. Biol. 217: 3656-3663. Manuscript waiting for release to PMC. This article was highlighted in Inside JEB.
2. `imo, L and Y. Park (2014) Neuropeptidergic control of the hindgut in the black-legged tick Ixodes scapularis. International Journal for Parasitology. 44: 819-826. With the cover page picture.PMC4175087
3. Ko?i, J., L. `imo, and Y. Park (2014) Autocrine/paracrine dopamine in the salivary glands of the blacklegged tick Ixodes scapularis J. Insect Physiology 62:39-45 PMC4175087
4. `imo, L., Ko?i, J., Kim, D., and Y. Park (2014) Invertebrate Specific D1-like Dopamine Receptor in Control of Salivary Glands in the Black-Legged Tick Ixodes scapularis. J. Comparative Neurology 522:2038-2052 PMC3997621
- Type:
Other
Status:
Published
Year Published:
2014
Citation:
12. Jiang, H., Z. Wei, R. J. Nachman, K. Kaczmarek, J. Zabrocki, and Y. Park (2014) Functional characterization of five different PRXamide receptors of the red flour beetle Tribolium castaneum with peptidomimetics and identification of agonists and antagonists. Peptides http://dx.doi.org/10.1016/j.peptides.2014.11.004.
13. Jiang, H., Z. Wei, R. J. Nachman, M. E. Adams, and Y. Park (2014) Functional phylogenetics reveals contributions of pleiotropic peptide action to ligand-receptor coevolution. Scientific Reports 4: 6840. doi:10.1038/srep06800
14. Kristopher, S., H. Jiang, J. Fu, T. W. Phillips, R. W. Beeman, and Y. Park (2014) The Tribolium castaneum cell line TcA: a new tool kit for cell biology. Scientific Reports 4: 6840. doi:10.1038/srep06840
15. Jiang, H, Z. Wei, R. J. Nachman, and Y. Park (2014) Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea. Peptides 53:243-249 PMC3989431
16. Jung, S-H, J-H Lee, H-S Chae, J-Y Seong, Y. Park, Z-Y Park, and Y-J Kim (2014) Identification of a novel insect neuropeptide, CNMa and its receptor. FEBS Letters 588: 2037-2041.
17. Kirkpatrick, K., A. T. Marshall, A. P. Smith, J. Koci, Y. Park (2014) Individual differences in impulsive and risky choice: Effects of environmental rearing conditions. Behavioral brain research 269C: 115-127.
18. Agrawal, S., M. Kelkenberg, K. Begum, S. Lea, C. E. Williams, K. J. Kramer, R. W. Beeman, Y. Park, S. Muthukrishnan, H. Merzendorfer (2014) Two essential peritrophic matrix proteins mediate matrix barrier functions in the insect midgut. Insect Biochem. Mol. Biol. 49:24-34.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
1. Park, Y. (2014) Mechanisms in control of tick salivary secretion. Purdue University, West Lafayette, IN
2. Park, Y. (2014) Mechanisms in control of tick salivary secretion. Baylor University, Waco, TX
3. Park, Y. (2014) Molecular evolution of neuropeptide signaling system. Wake Forest University, NC
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
1. Kim, D. A. and Y. Park (2014) Transcriptomics of tick salivary glands. Korean Young Entomologist Symposium in Entomological Society of America Annual Conference at Portland, OR.
2. Kim, D. A. and Y. Park (2014) Osmoregulatory functions of V-ATPase and Na/K-ATPase in the salivary gland of the blacklegged tick, Ixodes scapularis Say. Student competition in Entomological Society of America Annual Conference at Portland, OR
3. Urban, J. and Y. Park (2014) Molecular and functional characterization of a prostaglandin E2 synthase in the blacklegged tick, Ixodes scapularis Say. Student competition in Entomological Society of America Annual Conference at Portland, OR
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience: We made 6 publicaitons in the peer-reviewed international journals, 10 presentations in national conferences and in Universities at the national and international levels. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Two postdoctoral researcher and two graduate students were trained. · Ladislav Simo (Postdoctoral researcher) · Jurak Koci (Postdoctoral researcher) · Donghun Andy Kim (Ph.D. student) · Joshua Urban (Ph.D. student) How have the results been disseminated to communities of interest? We made 6 publicaitons in the peer-reviewed international journals, 10 presentations in national conferences and in Universities at the national and international levels. A part of research results were used in the class teaching "insect Physiology". What do you plan to do during the next reporting period to accomplish the goals? Current technical difficulties in doing RNA interference technique will be overcome by using the techniques for injecting pharmacological drugs into the individual salivary galnd acini.
Impacts
What was accomplished under these goals?
We have completed all three aims except the progresses being made in the Aim 3. We have identified the neuropeptides that controls the salivary glands as SIFamide and Myoinhibitory peptide (Aim 1). We have identified and characterized two different receptors for dopamine, SIFamide receptor, and MIP receptor expressed in the salivary glnads (Aim 2). The functions of the signaling system was investigated by using pharmacological tools. Unfortunately, however, RNA interference (RNAi) for functional studies found that the RNAi in tick species are tissue specific or gene specific, causing difficulties in achieving the proposed Aim 3. We have been developing a technique for in vitro microinjections of bioactive compounds in the acini to overcome this problem in the functional study. We have been developing a technique for in vitro microinjections of bioactive compounds in the acini to overcome this problem in the functional study.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Koci, J., L. Simo, and Y. Park (2013) Validation of internal reference genes for real-time quantitative PCR studies in the tick Ixodes scapularis. J. Medical Entomology, 50:79-84.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Simo, L., Koci, J., and Y. Park (2013) Receptors for the neuropeptides, myoinhibitory peptide and SIFamide, in control of the salivary glands of the blacklegged tick Ixodes scapularis. Insect Biochem. Mol. Biol., 43:376-387.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
4. Jiang, H., A. Ikhagva, I. Daubnerov�, H. Chae, L. `imo, S-H. Jung, Y-K. Yoon, N-R Lee, Y. J. Seong, D. Zitnan, Y. Park, and Y-J. Kim (2013) Natalisin, a new tachykinin-like signaling system, regulates sexual activity and fecundity in insects. Proc. Nat. Aca. Sci. USA. www.pnas.org/cgi/doi/10.1073/pnas.1310676110.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2013
Citation:
Jiang, H, Z. Wei, R. J. Nachman, and Y. Park (2013) Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea. Peptides (In press)
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2013
Citation:
Simo, L., Koci, J., Kim, D., and Y. Park (2013) Invertebrate specific D1-like dopamine receptor in control of salivary gland in the black-legged tick, Ixodes scapularis. J. Comparative Neurology (In press) DOI: 10.1002/cne.23515
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Koci, J., P. Ramaseshadri, R. Bolognesi, G. Segers, R. Flannagan, and Y. Park (2013) Ultrastructural changes caused by Snf7 RNAi in larval enterocytes of western corn rootworm (Diabrotica virgifera virgifera Le Conte). PLoS ONE 9(1): e83985. doi:10.1371/journal.pone.0083985.
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: The proposed activities are with three aims listed below. 1. Identifying candidate neuropeptides relevant to salivary functions. 2. Identifying the G protein-coupled receptors for dopamine, PGE2, MIP, and SIFamide. 3. Investigation of the functions of signaling systems in control of salivary secretion. We have completed the aim 1 and 2. Two manuscripts are being ready to be sent out for publications. We have identified immunoreactive neurons and their projections for more than 30 neuropeptide antibodies. MALDI analysis has uncovered more than 40 peaks for the molecular masses of predicted neuropeptides. Neuropeptide receptors (SIFamide receptor and MIP receptors) and dopamine receptors were identified and cloned. Unfortunately, however, RNA interference (RNAi) for functional studies found that the RNAi in tick species are tissue specific or gene specific, causing difficulties in achieving the proposed Aim 3. We are developing a technique for in vitro microinjections of bioactive compounds in the acini to overcome this problem in the functional study. PARTICIPANTS: A postdoctoral researcher was trained. Ladislav Simo TARGET AUDIENCES: We made three invited (the first 3) and 8 contributed presentations in local, national, and international conferences. 1. Park, Y. (2012) Molecular arthropod physiology in Park laboratory; current progresses and upcoming directions. Department of Entomology, Kansas State University. 2. Park, Y. (2012) Molecular physiology of tick salivary glands. International congress of Entomology 2012 Daegu, South Korea. 3. Park, Y. (2012) Molecular physiology of tick salivary secretion. Department of Entomology and Plant pathology, Oklahoma State University, Stillwater. 4. Urban, J. and Y. Park (2012) Prostaglandin E2 Synthase in the blacklegged tick, Ixodes scapularis. Kansas State Research Forum, K-State 5. Urban, J. and Y. Park (2012) Prostaglandin E2 Synthase in the blacklegged tick, Ixodes scapularis. North Central Branch meeting, Lincoln, NE, Entomological Society of America (Award 3rd place in Student oral presentation) 6. Urban, J. and Y. Park (2012) Prostaglandin E2 Synthase in the blacklegged tick, Ixodes scapularis. Entomological Society of America National Meeting, Knoxville, TN (Award 2nd place in Student Competition for the President's Prize) 7. Kim, D. A., Simo, L, and Y. Park (2012) Different physiological roles of two dopamine receptors in isolated salivary glands of the blacklegged tick, Ixodes scapularis" Entomological Society of America at Knoxville, TN, (Award 1st place in Student Competition for the President's Prize) 8. Kim, D. A., Simo, L, and Y. Park (2012) Different physiological roles of two dopamine receptors in isolated salivary glands of the blacklegged tick, Ixodes scapularis" Research and the State at Kansas State University 9. Kim, D. A., Simo, L, and Y. Park (2012) Different roles of two dopamine receptors, dopamine receptor and D1-like receptor, in control of salivary secretion in the blacklegged tick, Ixodes scapularis" GIST International Symposium for Invertebrate Neuropeptides and GPCRs, Gwangju, Korea 10. Kim, D. A., Simo, L, and Y. Park (2012) Different physiological roles of two dopamine receptors in isolated salivary glands of the blacklegged tick, Ixodes scapularis" International Congress of Entomology, Daegu, Korea 11. Kim, D. A., Simo, L, and Y. Park (2012) Different physiological roles of two dopamine receptors in isolated salivary glands of the blacklegged tick, Ixodes scapularis" North Central Branch of the Entomological Society of American at Lincoln, NE (Award 3rd place in Student Competition) PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We have completed a part of Specific Aim 1 and 2. A part of Specific Aim 3 is being explored. These accomplishments were presented in international, national, and local conferences and published in a peer reviewed journal. One publication and 11 presentations were made.
Publications
- We have made one publication in peer-reviewed journals in 2012.
- 1. Koci, J., Simo, L., and Y. Park (2012) Validation of internal reference genes for real-time quantitative PCR studies in the tick Ixodes scapularis. Journal of Medical Entomology, (in press).
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: The first aim of the study is to identify candidate neuropeptides relevant to salivary functions. We have partially achieved this goal by using immunohistochemistry of 15 different neuroepetides on tick synganglion and salivary gland. Among five antibodies resulting in the positive immunoreactivities, two of them were identified by using proteomics as SIFamide and myoinhibitory peptide (MIP), which was published in 2009. The immunohistochemistry led us to mapping the neuropeptidergic cells and their projections in the tick, allowing us to explore the anatomy-based functions. Currently, we established a frame of an on-line based searchable database named Tick Neuropeptide Atlas. This database will be further expanded to the proteomics data. We have identified and cloned the receptors for the neuropeptides in the salivary glands: MIP receptor and SIFamide receptor. Functional assay in CHO cell confirmed their activities upon the treatment of endogenous ligands MIP and SIFamide, respectively. Antibodies against these receptors have been raised and now used to perform immunohistochemistry to determine the locations of the receptor expressions. Two dopamine receptors expressed in the salivary glands were identified and cloned (Simo et al, 2011, PLos ONE). We hypothesize that two dopamine receptors have independent functions in the tick salivary secretion; one for fluid secretion and the other for control of the valve of each acinus. Physiological study is being performed to test this hypothesis. PARTICIPANTS: A postdoctoral researcher is being trained, Ladislav Simo. TARGET AUDIENCES: We made 6 invited (the first 6) and 2 contributed presentations in local, national, and international conferences : Park, Y. (2011) Signaling pathways for tick salivary secretion. Department of Biochemistry, University of Nevada, Reno. Park, Y. (2011) Signaling pathways for tick salivary secretion Signaling workshop in Entomological Society of America Annual Meeting in Reno, Nevada. Park, Y. (2011) Molecular physiology of tick salivary glands: How to spit. Department of Entomology, University of Georgia, Athens Park, Y. (2011) Neuropeptidergic control of the salivary gland in blacklegged tick Ixodes scapularis. Keynote speaker, International Symposium on Insect Physiology, Biochemistry, and Molecular Biology, Shanghi, China Park, Y. (2011) New insecticidal target: G protein-coupled receptors of insect. Monsanto, St. Louis Simo, L. and Park, Y. (2011) Two novel neuropeptides and their receptors in innervation of tick salivary glands. Society for Experimental Biology 2011 Annual main meeting, Glasgow, UK Koči, J., Simo, L., and Y. Park (2011) Catecholamine biosynthetic enzymes, tyrosin hydroxylase and dopadecarboxylase in the Ixodes scapularis. Poster presentation in 5th Arthropod Genomics Symposium 2011, Kansas City Simo, L., Koci, J., and Y. Park (2011) Myoinhibitory peptide and SIFamide receptors in neuropeptide signaling pathways controlling the tick salivary glands. Poster presentation in 5th Arthropod Genomics Symposium 2011 PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We have completed a part of Specific Aim 1 and a part of Specific Aim 2. A part of Specific Aim 3 has been explored. These accomplishments were presented in international, national, and local conferences and published in peer reviewed journal. Two publications and 8 presentations were made.
Publications
- 2. Simo, L., D. Zitnan, Y. Park (2011) Neural control of salivary glands in ixodid ticks. Journal of Insect Physiology. Early edition in doi:10.1016/j.jinsphys.2011.11.006
- 1. Simo, L., J. Koci, Z. Dusan, and Y. Park (2011) Evidence for D1 dopamine receptor activation by a paracrine signal of dopamine in tick salivary glands. PLoS ONE 6(1):e16158.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Immunohistochemistry of 15 different neuroepetides on tick synganglion and salivary gland found at least a total of five different antibodies stained the various neural projections reaching to the salivary glands (SGs). Peptidomics of the extracts for each synganglia and salivary gland revealed the components common for both tissue extracts. In the proteomics, we identified two major controller of the SGs, myoinhibitory peptide (MIP) and SIFamide. There is one more unidentified peptide revealed in the proteomics study, for which we continuously put our effort to identify the amino acid sequence and the gene. The genes encoding SIFamide and MIP have been cloned. We have cloned MIP and MIP receptors based on the homology searches of Ixodes genome. The composite clone for the receptor has been constructed in a mammalian cell expression vector and tested in a functional assay. In situ hybridization using the MIP cDNA as the probe found the immunoreactive cells PcSG were authentic cell expressing the MIP. In addition, we have done a parallel study on SIFamide and SIFamide receptors for gene cloning and in situ hybridization. In addition to the goals specified, we have contributed neuropeptide annotation on the VectorBase web site for Ixodes scapularis genome annotations. PARTICIPANTS: A postdoctoral researcher are being trained. Ladislav Simo TARGET AUDIENCES: We made over 6 invited/contributed presentations in local, national, and international conferences. Simo, L., Koci, J., and Y. Park (2010) Dopamine receptor D1 in control of the salivary glands in the blacklegged tick Ixodes scapularis. Poster presentation at the Entomological Society of America Annual meeting 2010, San Diego, California Koci, J., Simo, L., and Y. Park (2010) Biosynthesis of catecholamines in the salivary glands of tick: Tyrosine hydroxylase and Dopa decarboxylase. Poster presentation at the Entomological Society of America Annual meeting 2010, San Diego, California Simo, L., Koci, J., and Y. Park (2010) Dopamine receptor D1 in control of the salivary glands in the blacklegged tick Ixodes scapularis. Poster presentation at the Entomological Society of America Annual meeting 2010, San Diego, California Simo, L., Koci, J., and Y. Park (2010) Dopamine receptor D1in the salivary secretion of the blacklegged tick Ixodes scapularis. Poster presentation in 4th Arthropod Genomics Symposium 2010, Kansas City Koci, J., Simo, L., and Y. Park (2010) Searching the suitable internal control genes for real-time quantitative RT-PCR studies in Ixodes scapularis tick. Poster presentation in 4th Arthropod Genomics Symposium 2010, Kansas City Zitnan, D., L. Simo, L. Roller, I. Daubnerova, Y. Park, and M.E. Adams (2010) Complex peptidergic neuroendocrine network in ticks. Invertebrate Neuropeptide Conference 2010, Merida, Mexico PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We have completed a part of Specific Aim 1 and a part of Specific Aim 2. These accomplishments were presented in international, national, and local conferences and published in peer reviewed journal. One publication and 8 presentations were made.
Publications
- We have three publications in peer-reviewed journals in 2010 on this topic.
- Simo, L., J. Koci, Z. Dusan, and Y. Park (In press) 2010. Evidence for D1 dopamine receptor activation by a paracrine signal of dopamine in tick salivary glands. PLoS ONE
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: This is the first year of the 5-yr project. We have cloned MIP and MIP receptors based on the homology searches of Ixodes genome. Their 5' and 3'ends were identified by RACE (rapid amplification of cDNA ends), and composite clone for the receptor has been constructed in a mammalian cell expression vector. In situ hybridization using the MIP cDNA as the probe found the immunoreactive cells PcSG were authentic cell expressing the MIP. Currently functional expression of MIP receptor is in progress. As it was prepared in the pitfalls and alternatives in the original proposal, we found that the Ca++ mediated pathway is not activated in the expression of the MIP receptor, thus, we started an alternative approach for the functional expression. We use GloSensor (Promega) for detection of cAMP production. PARTICIPANTS: A postdoctoral researcher is being trained. Ladislav Simo TARGET AUDIENCES: We made over 6 invited/contributed presentations in local, national, and international conferences. Park, Y. (2009) Neuropeptidergic control of the salivary gland in blacklegged tick Ixodes scapularis. Entomology and Emerging Pathogen Institute in University of Florida. Park, Y. (2009) Neural control of the salivary gland in blacklegged tick. Seoul National University, Korea. Park, Y. (2009) Neuropeptidergic control of the salivary gland in blacklegged tick Ixodes scapularis. Diagnostic medicine/Pathology seminar series in Kansas State University. Simo, L., and Y. Park (2009) To novel neuropeptides in innervation of tick salivary glands: myoinhibitory peptide and SIFamide. The 6th Asia-Pacific Congress of Entomology, Beijing, China. Simo, L., Y. Park, and Z. Dusan (2009) Peptidergic innervation of salivary glands of the black-legged tick Ixodes scapularis (Say 1821). VIth International Conference on Arthropods Chemical Physiological, Biotechnological and Environmental Aspects, Ochotnica Dolna, Poland. Simo, L. and Y. Park (2009) Colocalization of two novel neuropeptides in innervation of salivary gland of the black legged tock Ixodes scapulartis. Poster presentation in 3rd Arthropod Genomics Symposium 2009, Kansas City. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We will expand the study of the roles of MIP and MIP receptor in the tick salivary secretion. The fundamental knowledge obtained from the MIP funciton is eventually expected to lead to the design of compounds and vaccines to prevent disease transmissions by ticks. Furthermore, this study can be extended to other tick species that have large negative impacts on the economy and health.
Publications
- Ladislav R., I. Zitanova, L. Dai, L. Simo, Y. Park, H. Satake, Y. Tanaka, M. E. Adams, and D. Zitnan. (2009) (In press) Ecdysis triggering hormone signaling in arthropods. Peptides
- Simo, L., D. Zitnan, and Y. Park (2009) Two novel neuropeptides in innervation of the salivary glands of the blacklegged tick Ixodes scapularis: Myoinhibitory peptides and SIF amide. Journal of Comparative Neurology. 517: 551-563.
- Simo, L., M. Slovak, Y. Park and D. Zitnan (2009) Identification of a complex peptidergic neuroendocrine network in the hard tick, Rhipicephalus appendiculatus. Cell and Tissue Research 335: 639-655.
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