A "Four Core Genotypes" rat model to distinguish mechanisms underlying sex-biased phenotypes and diseases.

Background: We have generated a rat model similar to the Four Core Genotypes mouse model, allowing comparison of XX and XY rats with the same type of gonad. The model detects novel sex chromosome effects (XX vs. XY) that contribute to sex differences in any rat phenotype. Methods: XY rats were produced with an autosomal transgene of Sry , the testis-determining factor gene, which were fathers of XX and XY progeny with testes. In other rats, CRISPR-Cas9 technology was used to remove Y chromosome factors that initiate testis differentiation, producing fertile XY gonadal females that have XX and XY progeny with ovaries. These groups can be compared to detect sex differences caused by sex chromosome complement (XX vs. XY) and/or by gonadal hormones (rats with testes vs. ovaries). Results: We have measured numerous phenotypes to characterize this model, including gonadal histology, breeding performance, anogenital distance, levels of reproductive hormones, body and organ weights, and central nervous system sexual dimorphisms. Serum testosterone levels were comparable in adult XX and XY gonadal males. Numerous phenotypes previously found to be sexually differentiated by the action of gonadal hormones were found to be similar in XX and XY rats with the same type of gonad, suggesting that XX and XY rats with the same type of gonad have comparable levels of gonadal hormones at various stages of development. Conclusion: The results establish a powerful new model to discriminate sex chromosome and gonadal hormone effects that cause sexual differences in rat physiology and disease.

FraidyRat: A Virtual Module Examining the Neural Circuitry Underlying Fear Conditioning.

FraidyRat is a teaching tool that allows students to investigate the neural basis of fear conditioning and extinction using a virtual rat with a virtual brain. FraidyRat models well-known phenomena at both a behavioral and neural level. Students use virtual versions of tract tracing, systemic and intracerebrally infused drugs, neural recording, and electrical stimulation to understand the neural substrates underlying the observed behavior. This module helps students develop critical thinking skills in order to deduce immediate cause and effect as well as inductive reasoning to grasp the broader scheme. This module utilizes scaffolded instruction and formative assessment to shape the thinking of students as they unfold and discover the neural mechanisms responsible for fear conditioning and extinction in FraidyRat, which largely reflect what is found in real rats. Experience with this three-week module resulted in students showing significant gains in content knowledge as well as a trend toward gains in critical thinking. An attitudinal questionnaire showed that students had an overall positive experience. This module can be replicated at any institution with just a computer. All materials are available at: https://mdcune.psych.ucla.edu/modules/fraidy-rat.

PROJECT DiViNe: DIVERSE VOICES IN NEUROSCIENCE: Profiles of Rita Levi-Montalcini, Ricardo Miledi, Simon LeVay, Erich Jarvis, and Steve Ramirez.

In this paper we share the first five of what we hope will be many profiles of neuroscientists from historically underrepresented or marginalized groups. This initial collection of profiles, meant to stake out the general territory for future offerings, takes as its subjects a fairly broad range of individuals from Nobel laureates to early career scientists and educators. The goal of this project is to facilitate the dissemination of materials neuroscience educators can use to highlight the scientific contributions and personal stories of scientists from historically marginalized groups, and has been developed more extensively in the Editorial that accompanies this collection (Frenzel and Harrington, 2021). We believe that by sharing these stories, and highlighting the diversity of those who have and will continue to contribute to the field of neuroscience, we can help to foster a more inclusive discipline for our undergraduate students. Each of these profiles is a testament to the respect these contributors hold for their subjects. We hope that others might see this new feature as an opportunity to share the admiration they have for those who have impacted them as colleagues, mentors, and role models.

Teaching Neuroscience: Reviving Neuroanatomy, Notes on the 2022 Society for Neuroscience Professional Development Workshop on Teaching.

Students often find neuroanatomy a daunting exercise of rote memorization in a dead language. This workshop was designed to enliven the teaching of neuroanatomy. We recast the topic by extending it to the cellular and sub-cellular levels, animating it by learning to build a brain, and infusing the topic with the lively arts. Due to COVID's interference with the usual schedule of Society for Neuroscience (SfN) events, the 2021 Professional Development Workshop on Teaching was held as a webinar on April 12, 2022 with a follow-up question and answer session on June 7. In this workshop, not only were innovative teaching methods presented, but also the very definition of neuroanatomy was pushed to the limits-even reaching into the molecular and subcellular level. The presenters provided means of engaging students that were no cost, low cost, or well within the reach of most academic institutions. Judging by the attendance, this webinar was quite successful in its goals. Our speakers presented exciting and varied approaches to teaching neuroanatomy. Kaitlyn Casimo presented how the vast resources of the Allen Institute could be employed. Marc Nahmani described how open data resources could be utilized in creating a Course-Based Undergraduate Research Experience (CURE) on neural microanatomy. Erika Fanselow presented novel ways to overcome one of students' big hurdles in grasping neuroanatomy: understanding 3-D relationships. Len White described a creative approach in teaching neuroanatomy by incorporating the humanities, particularly art and literature. This article presents synopses of the presentations, which are written by the four presenters. Additionally, prompted by questions from the viewers, we have constructed a table of our favorite resources. A video of the original presentations as well as links to the subsequent Q & A sessions is available at https://neuronline.sfn.org/training/teaching-neuroscience-reviving-neuroanatomy/.

Teaching Computation in Neuroscience: Notes on the 2019 Society for Neuroscience Professional Development Workshop on Teaching.

The 2019 Society for Neuroscience Professional Development Workshop on Teaching reviewed current tools, approaches, and examples for teaching computation in neuroscience. Robert Kass described the statistical foundations that students need to properly analyze data. Pascal Wallisch compared MATLAB and Python as programming languages for teaching students. Adrienne Fairhall discussed computational methods, training opportunities, and curricular considerations. Walt Babiec provided a view from the trenches on practical aspects of teaching computational neuroscience. Mathew Abrams concluded the session with an overview of resources for teaching and learning computational modeling in neuroscience.

Brain Volume Fractions in Mammals in Relation to Behavior in Carnivores, Primates, Ungulates, and Rodents.

The volume fraction (VF) of a given brain region, or the proper mass, ought to reflect the importance of that region in the life of a given species. This study sought to examine the VF of various brain regions across 61 different species of mammals to discern if there were regularities or differences among mammalian orders. We examined the brains of carnivores (n = 17), ungulates (n = 8), rodents (n = 7), primates (n = 11), and other mammals (n = 18) from the online collections at the National Museum of Health and Medicine. We measured and obtained the VF of several brain regions: the striatum, thalamus, neocortex, cerebellum, hippocampus, and piriform area. We refined our analyses by using phylogenetic size correction, yielding the corrected (c)VF. Our groups showed marked differences in gross brain architecture. Primates and carnivores were divergent in some measures, particularly the cVF of the striatum, even though their overall brain size range was roughly the same. Rodents predictably had relatively large cVFs of subcortical structures due to the fact that their neocortical cVF was smaller, particularly when compared to primates. Not so predictably, rodents had the largest cerebellar cVF, and there were marked discrepancies in cerebellar data across groups. Ungulates had a larger piriform area than primates, perhaps due to their olfactory processing abilities. We provide interpretations of our results in the light of the comparative behavioral and neuroanatomical literature.

Using Online Images to Teach Quantitative Skills via Comparative Neuroanatomy: Applying the Directives of Vision and Change.

Vision and Change calls for increasing the quantitative skills of biology majors, which includes neuroscience majors. Accordingly, we have devised a module to give students practice at regression analyses, covariance, and ANOVA. This module consists of a quantitative comparative neuroanatomy lab in which students explore the size of the hippocampus relative to the brain in 62 different mammalian species-from an anteater to a zebu. We utilize a digital image library (with appropriate metadata) allowing students to quantify the size of the hippocampus as well as obtain an index of the size of the brain in these various species. Students then answer the following questions: (1) Do brains scale with body size? (2) Does the hippocampus scale with brain size? (3) If we control for body size, does the hippocampus still scale with brain size? (4) How does the hippocampus change as a proportion of brain size? (5) Is the proportional scaling of the hippocampus different among primates, carnivores, and other mammals? (6) Do the data provide evidence for mosaic or concerted evolution? Measures of the pedagogical efficacy showed clear and significant gains on a PreTest vs PostTest assessment of material related to the module. An open ended qualitative measure revealed students' perception of the purposes of the module, which were consistent with the learning goals. This module utilizes open access digital resources and can be performed at any institution. All the materials or links to online resources can be found at https://mdcune.psych.ucla.edu/modules/cna.

Teaching with Big Data: Report from the 2016 Society for Neuroscience Teaching Workshop.

As part of a series of workshops on teaching neuroscience at the Society for Neuroscience annual meetings, William Grisham and Richard Olivo organized the 2016 workshop on "Teaching Neuroscience with Big Data." This article presents a summary of that workshop. Speakers provided overviews of open datasets that could be used in teaching undergraduate courses. These included resources that already appear in educational settings, including the Allen Brain Atlas (presented by Joshua Brumberg and Terri Gilbert), and the Mouse Brain Library and GeneNetwork (presented by Robert Williams). Other resources, such as NeuroData (presented by William R. Gray Roncal), and OpenFMRI, NeuroVault, and Neurosynth (presented by Russell Poldrack) have not been broadly utilized by the neuroscience education community but offer obvious potential. Finally, William Grisham discussed the iNeuro Project, an NSF-sponsored effort to develop the necessary curriculum for preparing students to handle Big Data. Linda Lanyon further elaborated on the current state and challenges in educating students to deal with Big Data and described some training resources provided by the International Neuroinformatics Coordinating Facility. Neuroinformatics is a subfield of neuroscience that deals with data utilizing analytical tools and computational models. The feasibility of offering neuroinformatics programs at primarily undergraduate institutions was also discussed.

Undergraduate Neuroscience Education in the U.S.: Quantitative Comparisons of Programs and Graduates in the Broader Context of Undergraduate Life Sciences Education.

The impact of undergraduate neuroscience programs on the broader landscape of life sciences education has not been described. Using data from the National Center for Education Statistics, we found that the number of undergraduate neuroscience programs in the U.S. continues to grow. Within any given institution, neuroscience programs exist alongside a small number of other life sciences undergraduate programs, suggesting that neuroscience is one of few major options from which students can choose from at many institutions. Neuroscience majors constitute a substantial proportion of all life sciences graduates at many institutions, and in several cases, neuroscience majors were the majority of life sciences graduates. Thus, neuroscience programs contribute substantially to life sciences education, and neuroscience is a highly attractive major among undergraduate students where these programs are available. These data have implications for institutions with existing neuroscience programs as well as for institutions seeking to establish a new program.

Proposed Training to Meet Challenges of Large-Scale Data in Neuroscience.

The scale of data being produced in neuroscience at present and in the future creates new and unheralded challenges, outstripping conventional ways of handling, considering, and analyzing data. As neuroinformatics enters into this big data era, a need for a highly trained and perhaps unique workforce is emerging. To determine the staffing needs created by the impending era of big data, a workshop (iNeuro Project) was convened November 13-14, 2014. Participants included data resource providers, bioinformatics/analytics trainers, computer scientists, library scientists, and neuroscience educators. These individuals provided perspectives on the challenges of big data, the preparation of a workforce to meet these challenges, and the present state of training programs. Participants discussed whether suitable training programs will need to be constructed from scratch or if existing programs can serve as models. Currently, most programs at the undergraduate and graduate levels are located in Europe-participants knew of none in the United States. The skill sets that training programs would need to provide as well as the curriculum necessary to teach them were also discussed. Consistent with Vision and Change in Undergraduate Biology Education: A Call to Action, proposed curricula included authentic, hands-on research experiences. Further discussions revolved around the logistics and barriers to creating such programs. The full white paper, iNeuro Project Workshop Report, is available from iNeuro Project.

An Instructor's Guide to (Some of) the Most Amazing Papers in Neuroscience.

Although textbooks are still assigned in many undergraduate science courses, it is now not uncommon, even in some of the earliest courses in the curriculum, to supplement texts with primary source readings from the scientific literature. Not only does reading these articles help students develop an understanding of specific course content, it also helps foster an ability to engage with the discipline the way its practitioners do. One challenge with this approach, however, is that it can be difficult for instructors to select appropriate readings on topics outside of their areas of expertise as would be required in a survey course, for example. Here we present a subset of the papers that were offered in response to a request for the "most amazing papers in neuroscience" that appeared on the listserv of the Faculty for Undergraduate Neuroscience (FUN). Each contributor was subsequently asked to describe briefly the content of their recommended papers, their pedagogical value, and the audiences for which these papers are best suited. Our goal is to provide readers with sufficient information to decide whether such articles might be useful in their own classes. It is not our intention that any article within this collection will provide the final word on an area of investigation, nor that this collection will provide the final word for the discipline as a whole. Rather, this article is a collection of papers that have proven themselves valuable in the hands of these particular educators. Indeed, it is our hope that this collection represents the inaugural offering of what will become a regular feature in this journal, so that we can continue to benefit from the diverse expertise of the FUN community.

ERIN: A Portal to Resources for Higher Education in Neuroscience.

ERIN, Educational Resources in Neuroscience, is the Society for Neuroscience's web portal to selected, high-quality materials for higher education. A Board of Editors approves resources after describing them and classifying them by topic, subtopic, media type, author, and appropriate educational level. Some resources are also accompanied by reviews and ratings from faculty who have used the resource. These features make a search of ERIN far more useful than a typical Google search. ERIN's development was funded by the National Science Foundation with a three-year grant to SfN. Along the way, various unexpected problems arose and solutions were found, many of which are described in this overview of ERIN's history and the various decisions that were made in its design and development.

Gel Scramble: An E-Tool for Teaching Molecular Neuroscience.

In this completely digital teaching module, students interpret the results of two separate procedures: a restriction endonuclease digestion, and a polymerase chain reaction (PCR). The first consists of matching restriction endonuclease digest protocols with images obtained from stained agarose gels. Students are given the sequence of six plasmid cDNAs, characteristics of the plasmid vector, and the endonuclease digest protocols, which specify the enzyme(s) used. Students calculate the expected lengths of digestion products using this information and free tools available on the web. Students learn how to read gels and then match their predicted fragment lengths to the digital images obtained from the gel electrophoresis of the cDNA digest. In the PCR experiment, students are given six cDNA sequences and six sets of primers. By querying NCBI BLAST, students can match the PCR fragments to the lengths of the predicted in silico PCR products. The ruse posed to students is that the gels were inadvertently mislabeled during processing. Although students know the experimental details, they do not know which gel goes with a given restriction endonuclease digest or PCR-they must deduce the answers. Because the gel images are from actual students' experiments, the data sometimes result from mishandling/mislabeling or faulty protocol execution. The most challenging part of the exercise is to explain these errors. This latter aspect requires students to use critical thinking skills to explain aberrant outcomes. This entire exercise is available in a digital format and downloadable for free at http://mdcune.psych.ucla.edu/modules/gel.

Quantitative examination of the bottlenose dolphin cerebellum.

Neuroanatomical research into the brain of the bottlenose dolphin (Tursiops truncatus) has revealed striking similarities with the human brain in terms of size and complexity. However, the dolphin brain also contains unique allometric relationships. When compared to the human brain, the dolphin cerebellum is noticeably larger. Upon closer examination, the lobule composition of the cerebellum is distinct between the two species. In this study, we used magnetic resonance imaging to analyze cerebellar anatomy in the bottlenose dolphin and measure the volume of the separate cerebellar lobules in the bottlenose dolphin and human. Lobule identification was assisted by three-dimensional modeling. We find that lobules VI, VIIb, VIII, and IX are the largest lobules of the bottlenose dolphin cerebellum, while the anterior lobe (I-V), crus I, crus II, and the flocculonodular lobe are smaller. Different lobule sizes may have functional implications. Auditory-associated lobules VIIb, VIII, IX are likely large in the bottlenose dolphin due to echolocation abilities. Our study provides quantitative information on cerebellar anatomy that substantiates previous reports based on gross observation and subjective analysis. This study is part of a continuing effort toward providing explicit descriptions of cetacean neuroanatomy to support the interpretation of behavioral studies on cetacean cognition.

Teaching neuroinformatics with an emphasis on quantitative locus analysis.

Although powerful bioinformatics tools are available for free on the web and are used by neuroscience professionals on a daily basis, neuroscience students are largely ignorant of them. This Neuroinformatics module weaves together several bioinformatics tools to make a comprehensive unit. This unit encompasses quantifying a phenotype through a Quantitative Trait Locus (QTL) analysis, which links phenotype to loci on chromosomes that likely had an impact on the phenotype. Students then are able to sift through a list of genes in the region(s) of the chromosome identified by the QTL analysis and find a candidate gene that has relatively high expression in the brain region of interest. Once such a candidate gene is identified, students can find out more information about the gene, including the cells/layers in which it is expressed, the sequence of the gene, and an article about the gene. All of the resources employed are available at no cost via the internet. Didactic elements of this instructional module include genetics, neuroanatomy, Quantitative Trait Locus analysis, molecular techniques in neuroscience, and statistics-including multiple regression, ANOVA, and a bootstrap technique. This module was presented at the Faculty for Undergraduate Neuroscience (FUN) 2011 Workshop at Pomona College and can be accessed at http://mdcune.psych.ucla.edu/modules/bioinformatics.

Using digital images of the zebra finch song system as a tool to teach organizational effects of steroid hormones: a free downloadable module.

Zebra finch song behavior is sexually dimorphic: males sing and females do not. The neural system underlying this behavior is sexually dimorphic, and this sex difference is easy to quantify. During development, the zebra finch song system can be altered by steroid hormones, specifically estradiol, which actually masculinizes it. Because of the ease of quantification and experimental manipulation, the zebra finch song system has great potential for use in undergraduate labs. Unfortunately, the underlying costs prohibit use of this system in undergraduate labs. Further, the time required to perform a developmental study renders such undertakings unrealistic within a single academic term. We have overcome these barriers by creating digital tools, including an image library of song nuclei from zebra finch brains. Students using this library replicate and extend a published experiment examining the dose of estradiol required to masculinize the female zebra finch brain. We have used this library for several terms, and students not only obtain significant experimental results but also make gains in understanding content, experimental controls, and inferential statistics (analysis of variance and post hoc tests). We have provided free access to these digital tools at the following website: http://mdcune.psych.ucla.edu/modules/birdsong.

Teaching bioinformatics and neuroinformatics by using free web-based tools.

This completely computer-based module's purpose is to introduce students to bioinformatics resources. We present an easy-to-adopt module that weaves together several important bioinformatic tools so students can grasp how these tools are used in answering research questions. Students integrate information gathered from websites dealing with anatomy (Mouse Brain Library), quantitative trait locus analysis (WebQTL from GeneNetwork), bioinformatics and gene expression analyses (University of California, Santa Cruz Genome Browser, National Center for Biotechnology Information's Entrez Gene, and the Allen Brain Atlas), and information resources (PubMed). Instructors can use these various websites in concert to teach genetics from the phenotypic level to the molecular level, aspects of neuroanatomy and histology, statistics, quantitative trait locus analysis, and molecular biology (including in situ hybridization and microarray analysis), and to introduce bioinformatic resources. Students use these resources to discover 1) the region(s) of chromosome(s) influencing the phenotypic trait, 2) a list of candidate genes-narrowed by expression data, 3) the in situ pattern of a given gene in the region of interest, 4) the nucleotide sequence of the candidate gene, and 5) articles describing the gene. Teaching materials such as a detailed student/instructor's manual, PowerPoints, sample exams, and links to free Web resources can be found at http://mdcune.psych.ucla.edu/modules/bioinformatics.

X chromosome number causes sex differences in gene expression in adult mouse striatum.

Previous research suggests that sex differences in the nigrostriatal system are created by direct effects of the sex chromosomes (XX vs. XY), independent of the action of gonadal hormones. Here we tested for sex chromosome effects on expression of three mRNAs in the striatum and nucleus accumbens of adult mice of the four core genotypes model (XX and XY gonadal males, XX and XY gonadal females). Mice were gonadectomized (GDX) at 47-51 days old to eliminate group differences in the levels of gonadal steroids. Three weeks later, mice were killed and brains collected for in situ hybridization of the striatum, or the striatum was dissected out for quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Expression in XX and XY mice was measured by in situ hybridization using riboprobes encoding the dynorphin precursor Pdyn (prodynorphin), the substance P precursor Tac1 (preprotachykinin) or dopamine D2 receptor. XX mice had higher expression, relative to XY mice of the same gonadal sex, of Pdyn and Tac1 mRNA in specific striatal regions. Quantitative PCR confirmed that GDX XX mice have higher Pdyn expression in striatum than XY mice, regardless of their gonadal sex. XX had higher Pdyn expression than XY or XO mice, indicating that the sex chromosome effect is the result of XX vs. XY differences in the number of X chromosomes, probably because of sex differences in the expression of X gene(s) that escape inactivation. We detected no sex chromosome effect on D2 receptor mRNA.