Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Laura Vicente-Rodríguez - University of Puerto Rico, Medical Sciences Campus
Co-Author(s): Nuria Vázquez and Mark Miller, Institute of Neurobiology, San Juan Solymar Rolón, University of Pennsylvania, Philadelphia Joshua Rosenthal, Marine Biological Laboratory, Massachusetts
Schistosomiasis, ‘bilharzia’ or ‘snail fever’ is a major global concern. It is the second most prevalent parasitic disease and it may affect 240 million people annually. Schistosoma mansoni is one of the trematode species that causes the most widespread form of intestinal schistosomiasis, which may result in abdominal bleeding, hepatosplenomegaly and intestinal damage. Important developmental stages of the Schistosoma mansoni life cycle occur within fresh water snails, specifically from the genus Biomphalaria, the parasite’s primary intermediate host. Neuropeptides act on specific receptors to regulate a wide range of cellular functions such as cardiac activity, synaptic transmission and locomotion. Accordingly, their receptors are promising molecular targets for pesticides and parasiticide drug development. In a recent study, the organization and immunohistochemical localization of two B. glabrata FMRFamide precursors was described. However, the role of this peptide in the B. glabrata’s neural signaling remains unknown. We hypothesize that a FMRFamide gated receptor triggers a neural response involved in B. glabrata’s vital functions such as feeding and sexual behavior and that disruption of this receptor could reduce the snail’s ability to survive. Using transcriptome data, full-length cDNAs for a FMRF-amide receptor were amplified by RT-PCR, cloned and sequenced. RNA from these clones was synthesized in vitro and injected in Xenopus laevis oocytes. Protein expression in oocytes was monitored through western blot and channel’s properties were characterized using electrophysiology. The resulting peptide gated channel physiological properties are in agreement with those described in the literature. In future work, the specific location of neurons encoding for this channel expression in B. glabrata will be determined by in-situ hybridization. This study aims to confirm the available data supporting the participation of FMRF-amide in the regulation of vital physiological and behavioral processes in gastropods and will explore the FMR-amide peptide signaling as target for snail control.
Funder Acknowledgement(s): This research is supported by: NSF HRD-1137725- CREST-Puerto Rico Center for Environmental Neuroscience and MBRS-RISE- R25GM061838.
Faculty Advisor: Mark Miller, email@example.com
Role: The parts that I have done from this research are: 1. Biomphalaria central nervous system dissection/isolation; 2. RNA extraction and cDNA synthesis; 3. DNA cloning by: RT-PCR amplification and Cell Transformation; 4. RNA invitro synthesis; 5. RNA injections in Xenopus oocytes; 6. Western Blot; 7. 2 electrodes voltage clamp recordings.