This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).
Funds from the Major Research Instrumentation-Recovery and Reinvestment (MRI-R2) program were provided to support the acquisition of a new transmission electron microscope (TEM) to replace a 30 year old TEM at the University of Puerto Rico Medical Sciences Campus (UPR-MSC). The TEM will be installed at the Institute of Neurobiology. The Institute is an interdisciplinary, interdepartmental research facility devoted to the study of the structure and function of the nervous system at all levels of organization from whole animal behavior to molecular biology. Because it is managed as a shared resource, the new TEM significantly enhances the research efforts of investigators at the Institute, at other campuses of the UPR, and at other universities in Puerto Rico. The projects benefitting from the TEM address the broad objectives of understanding basic mechanisms of connectivity of the nervous system, focusing on changes during development, after injury, and in response to the environment. The acquisition of the TEM also positively impacts training of graduates and medical students, undergraduates and science teachers. Courses in basic and advanced electron microscopy enrich graduate programs and increase the number of qualified users on the island. Community outreach efforts, which include workshops in electron microscopy for high school teachers and visits to the new facility by groups of high school students, also broaden the impact of the instrumentation upon science education. Following in the tradition of the UPR as a major contributor to the education and training of scientists, the shared electron microscope is instrumental in the training of future academic leaders in the field of Neuroscience. Results from the studies enabled by the new TEM will be disseminated by student and faculty presentations at regional and national meetings, and through publication in peer-reviewed journals.
PI: Rosa E. Blanco Awardee: Institute of Neurobiology, University of Puerto Rico Award Number: 0959225 The Institute of Neurobiology has a new electron microscope JEOL JEM 1011 installed and working. It is currently used by the investigators of the Institute. The scope is being used by several laboratories to study different characteristics of the nervous system, using simple models to understand basic mechanisms of survival after injury, nerve regeneration, connectivity between cells, and the molecular basis of addiction and behavior. Also Dr. Rosa Blanco teaches a graduate course on basic and advanced techniques for electron microscopy; this course includes a practical component where the students have access to the electron microscope. A total of six graduate students from the doctoral program of the Anatomy, Neurobiology and Microbiology departments participated in the course this year. The objective is to increase the capability of these students to apply this technique and use the instrument in their research projects. Examples of the research projects: from Dr Rosa Blanco’s laboratory Giam Vega, a graduate student, is studying the effect of neurotrophins on the speed and number of regenerating axons. He has found that ciliary neurotrophic factor and basic fibroblast factor increase the speed and number of regenerating axons, and with the electron microscope he has been able to show also a differential distribution of the regenerating axons with a larger number regenerating peripherally alongside the astrocytes. Mildred Duprey, a graduate student, is studying the effect of altering the concentration of brain derived neurotrophic factor on the number of synapses between retinal axons and tectal neurons after manipulation of tectal levels of neurotrophins; she is carrying out an electron microscopic analysis of thin sections to study synaptic formation after regeneration. Dr Jonathan Blagburn is studying the changes in the specificity of patterns of synaptic connections between sensory neurons and their targets with overexpression or knockout of the transcription factor Engrailed in Drosophila melanogaster. Currently his laboratory is using the electron microscope to establish the general pattern of monosynaptic connections of the engrailed positive cells of the Johnston’s Organ that connect with the giant fiber neuron in the auditory neuropil. They are interested in determining the changes that occur when Engrailed expression is perturbed in these sensory neurons. Dr Mark Miller has started a project studying biogenic amines in freshwater snail Biomphalaria glabrata, the primary intermediate host of the trematode Schistosoma mansoni that causes the disease schistosomiasis. He is interested in the ultrastructure of cells that stain with tyrosine hydroxylase in the nervous system (CNS) and peripheral tissues of B. glabrata. There is particular interest in the interactions of these cells with epithelial cells of the anterior foot and body wall; these cells could have relevance in nociceptive, or inflammation responses. Ana Ortiz from the laboratory of Dr. Maria Sosa is studying the ultrastructural localization of the receptors for tyramine/octopamine in the central nervous system of the freshwater prawn Macrobrachium rosenbergii. There are three morphotypes of this animal, each displaying different levels of aggression, and a comparatively higher expression level of these receptors has been reported in the more submissive animals. So far she has localized the receptors in a subpopulation of cerebral ganglion neurons and further immunotechniques are going to be used to compare the distribution and abundance of these receptors. Dr Garrett Searle from Dr Steven Treistman’s laboratory has shown that large conductance calcium- and voltage-activated potassium (BK) channels are potentiated by acute ethanol exposure, while longer durations of ethanol exposure result in molecular adaptation of the channel, altering neuronal sensitivity to ethanol. Ethanol exposure reduced synaptic enrichment of the BK channel alpha subunit. Interestingly, synapses on distal dendrites exhibited a greater degree of reduction of BK channel localization within synaptic compartments as compared to synapses on proximal dendrites. In collaboration with Dr Blanco he is currently developing a immunogold labeling technique to study at the electron microscopic level the changes of these channels in distal and proximal dendrites of hippocampal cultured neurons. As a direct result of learning the techniques three graduate students are currently having research projects for a specific topic that will be included in their dissertation. The PI has made specific efforts to contact two science teachers from local schools to organize visits from high school students to the electron microscope facility. The students were very enthusiastic because the visit was coordinated with their receiving basic cell biology lectures and identifying those structures with the microscope. At least two of those students are interested in doing summer research to further their knowledge of microscopy.
