In the Research core, interns dive into an immersive research experience that allows them to formulate their own projects on the neurobiology of ASD within a team-based and peer-mentoring training environment.
Interns form research teams to construct scientific projects within one of four research modules (Behavior, Functional, Cell Biology, Genomics). In brainstorming sessions, they build scientific premises and identify unknowns in the field to generate hypotheses and design experiments. Training addresses techniques and methods, data treatment and analyses as well as scientific writing, and design of research presentations.
The main outcome of the Research core is for interns to gain enduring and applicable scientific skills together with a heightened interest in neuro-developmental disorders research.
REACH Research Core.
Interns form teams to work in one of four research modules.
Each team receives training to formulate a hypothesis and design a research project to address relevant ASD-related gaps in knowledge under the guidance of their module’s faculty and student mentors.
Progress reports are presented weekly. The Summer Course ends with a Graduation Ceremony where Teams present their projects.
During the summer experience, interns form research teams. The team-based strategy is directed to create an environment in which interns learn collaborative problem solving, teamwork, leadership skills and conflict resolution. Faculty and student mentors guide team members to collectively learn to develop a scientific project, coordinate efforts, assign tasks, establish collaborations with other teams and effectively communicate their research.
REACH Research Modules & Teams
By the end of the Lecture core, interns are introduced to lab demonstrations for the techniques and methodologies utilized in each Research Module: Behavioral, Functional, Cell Biology (or Cellular) and Genomic. Presentations from faculty and students mentors familiarize interns with the kinds of questions each particular module can address, their advantages and disadvantages, and how they complement with other modules. Interns then rank modules based on their interest in the approach and the kinds of questions they are interested to address for potential scientific projects. Faculty allocate interns in Research Teams based on their ranking while maintaining the 2 high school-to-2 college students ratio.
Mentors from each module train, guide and supervise their team of interns to tackle an ASD-related question. Instruction starts with brainstorming sessions to review literature, formulate a hypothesis and design experiments. Interns are trained in the methodology while generating pilot data to assess the feasibility of the proposed project. Interns will receive specific instruction on keeping detailed records of protocols and experiments in their laboratory notebooks.
As interns brainstorm their projects, opportunities for collaboration projects often arise among research teams. Similarly, as work progresses, teams may naturally find themselves establishing new collaborations. If so, mentors of the potentially collaborative teams will assess the feasibility of the collaborative effort and work together with the teams to establish strategies and policies to carry out the planned experiments.
REACH Research Modules & Teams.
Interns form teams to work in one of four research modules. Each research module focuses on Behavioral, Functional, Cellular and Genomics studies, respectively.
Each team is made up of 4 interns (two high school and two college students) and faculty and student mentors.
REACH Behavioral Module
Behavior can be defined as the output of the nervous system’s function. Therefore, behavioral methods allow the quantification of a subject’s reaction to conditions placed on them in a controlled environment in order to shed light on its underlying neurobiological processes. Projects within the Behavioral module measure indexes of animal conduct to determine phenotypes of interest between an ASD rodent model and the corresponding controls.
Rodents are social creatures. Brain regions associated with social behavior include, the prefrontal cortex, anterior cingulate gyrus, amygdala and hippocampus (Hui et al., 2020-30470595). It has been reported that children with ASD have a lower density of connections amongst these regions, which are collectively known as the limbic system and that are associated with emotional processing (Namburi et al., 2016-PMID: 26647973). In the laboratory setting, social behavior can be inferred by quantifying the number of prosocial behaviors that occur when a laboratory animal is placed in an arena with another peer. A prosocial behavior refers to any interaction related to gaining social information. In rodents, prosocial behaviors include touching, sniffing, licking, and playing with cage-mates (Kim et al., 2021-PMID: 34091136).
Within the Behavioral module, interns can examine prosocial behaviors within an environment that can be manipulated into different social contexts, along with measures of basal locomotion and anxiety. Interns can also measure social communication via recording of ultrasonic vocalizations (USV), observed in pups after brief maternal separation (Hodges et al., 2017-PMID: 2855259).
PhD candidate Neuroscience, DHSU
Diana Dow-Edwards PhD
Prof. Physiology & Pharmacology, DHSU
John Kubie PhD
Assoc. Prof. Cell Biology, DHSU
REACH Functional Module
In the Functional module, interns use electrophysiological methods to measure the electrical activity of neurons in order to address questions about ASD pertinent to neuronal connectivity.
ASD is characterized by improper neural network connectivity across the brain (Kotila et al., 2021-PMID: 33206471). Areas compromised are the limbic system (prefrontal cortex, anterior cingulate gyrus, amygdala and hippocampus) and basal ganglia, a group of brain structures that contribute to motor control (Kilroy et al., 2019-PMID: 30925819). Communication between neurons depends on their electrical properties; precisely, the input synaptic activity into a neuron and its output action potential firing. Correct input-output is instrumental to proper transfer of information, and deficits in it have been linked to the ASD condition (Micheau et al., 2014-PMID: 24753134).
Projects within the Neuronal function module assess input-output properties between an ASD animal model and the corresponding controls. Interns can examine neuronal function in brain slices cut in a fashion that could contain the areas and connections of interest (e.g. different areas of the limbic system). Microelectrodes are then placed at particular parts of a neuron (e.g. dendrites, cell body) within the areas of interest to examine synaptic or action potential responses.
Juan Marcos Alarcon PhD
Assoc. Prof. Pathology, DHSU
REACH Cell Biology Module
In the Cell Biology module, interns use immunolabeling methods and confocal imaging to address questions about ASD pertinent to the identification of macromolecules of interest, be it protein, carbohydrate or nucleic acid, and assess their expression within specific cells or brain areas.
Alteration of factors that affect the development and morphology of neurons are linked with neurodevelopmental disorders, including ASD (Ryabinin, 2022-PMID: 34891220). Candidate factors under investigation include molecules that are important for neuronal development, such as immediate early genes c-Fos and Arc, and neural communication such as synaptic receptors NR1/2 and GluR1/2 (Bourgeron, 2015-PMID: 26289574). Also candidates are the translation regulator FMRP (Fragile X syndrome protein) and downstream products Shank and Homer, scaffolding proteins that play a critical role in synaptic activity (Soler et al., 2018-PMID: 29947605).
Projects within the Cell Biology module investigate mechanistic alterations in an ASD animal model compared to corresponding controls. Interns within the module can design projects to examine factors of interest along the transcriptional and translational machinery, and downstream products that modulate synaptic function.
Ivan A. Hernandez PhD
Director of the Imaging Facility, Asst. Prof. of Pathology, DHSU
MD/PhD candidate Neuroscience, DHSU
REACH Genomics Module
In the Genomics module, interns use transcriptome data analysis by means of RNA sequencing and PCR of targeted genes to address questions about altered gene expression profiles in ASD.
Genetics studies of ASD have identified several risk genes. Many of the risk genes encode proteins that can affect neuronal communication in the brain (synaptic plasticity): synaptic receptors, scaffolding proteins, cell adhesion molecules as well as proteins that are involved in chromatin remodeling, transcription, translation, degradation and cytoskeleton dynamics (Bourgeron 2015-PMID: 26289574).
Projects within the Genomics module use RNA sequencing methods and real-time PCR determinations of targeted genes to examine the transcript profile divergence in the expression of genes between an ASD animal model and controls. Interns carry out large-scale gene expression profile analyses (bio-informatics analysis) in conjunction with the mentors. Interns can carry out comparisons between or within specific cell types or brain regions of interest (e.g. limbic system, basal ganglia) to identify expression changes of genes whose products constitute mechanisms for transcription, translation and synaptic function.
Oleg Evgrafov PhD
Director, Human Genomics, Prof. of Cell Biology, DHSU