Illustrated Neuroanatomy of Ctenophores: Immunohistochemistry.

Ctenophores or comb jellies are representatives of an enigmatic lineage of early branching metazoans with complex tissue and organ organization. Their biology and even microanatomy are not well known for most of these fragile pelagic and deep-water species. Here, we present immunohistochemical protocols successfully tested on more than a dozen ctenophores. This chapter also illustrates neural organization in several reference species of the phylum (Pleurobrachia bachei, P. pileus, Mnemiopsis leidyi, Bolinopsis microptera, Beroe ovata, and B. abyssicola) as well as numerous ciliated structures in different functional systems. The applications of these protocols illuminate a very complex diversification of cell types comparable to many bilaterian lineages.

Comparison of histological delineations of medial temporal lobe cortices by four independent neuroanatomy laboratories.

The medial temporal lobe (MTL) cortex, located adjacent to the hippocampus, is crucial for memory and prone to the accumulation of certain neuropathologies such as Alzheimer's disease neurofibrillary tau tangles. The MTL cortex is composed of several subregions which differ in their functional and cytoarchitectonic features. As neuroanatomical schools rely on different cytoarchitectonic definitions of these subregions, it is unclear to what extent their delineations of MTL cortex subregions overlap. Here, we provide an overview of cytoarchitectonic definitions of the entorhinal and parahippocampal cortices as well as Brodmann areas (BA) 35 and 36, as provided by four neuroanatomists from different laboratories, aiming to identify the rationale for overlapping and diverging delineations. Nissl-stained series were acquired from the temporal lobes of three human specimens (two right and one left hemisphere). Slices (50 µm thick) were prepared perpendicular to the long axis of the hippocampus spanning the entire longitudinal extent of the MTL cortex. Four neuroanatomists annotated MTL cortex subregions on digitized slices spaced 5 mm apart (pixel size 0.4 µm at 20× magnification). Parcellations, terminology, and border placement were compared among neuroanatomists. Cytoarchitectonic features of each subregion are described in detail. Qualitative analysis of the annotations showed higher agreement in the definitions of the entorhinal cortex and BA35, while the definitions of BA36 and the parahippocampal cortex exhibited less overlap among neuroanatomists. The degree of overlap of cytoarchitectonic definitions was partially reflected in the neuroanatomists' agreement on the respective delineations. Lower agreement in annotations was observed in transitional zones between structures where seminal cytoarchitectonic features are expressed less saliently. The results highlight that definitions and parcellations of the MTL cortex differ among neuroanatomical schools and thereby increase understanding of why these differences may arise. This work sets a crucial foundation to further advance anatomically-informed neuroimaging research on the human MTL cortex.

A well-preserved cranium from the Judith River Formation (Montana, USA) reveals the inner ear and neuroanatomy of a Campanian baenid turtle.

A cranium belonging to a baenid turtle was recently recovered from the lower half of the Judith River Formation, Montana. Badlands Dinosaur Museum (BDM) 004 is a well-preserved partial cranium that includes the posterior cranial vault, cranial base, and otic capsules. Based on diagnostic characters, the skull can be attributed to Plesiobaena antiqua, which has been previously reported from the Judith River Formation. It also shares with palatobaenines projecting posterior processes of the tubercula basioccipitale and a prominent condylus occipitalis with a deep central pit, demonstrating variation within the Pl. antiqua hypodigm. In a phylogenetic analysis, an operational taxonomic unit of BDM 004 was positioned within Baenodda in an unresolved polytomy with Pl. antiqua, Edowa zuniensis, Palatobaeninae, and Eubaeninae. Microcomputed tomographic (µCT) scans revealed morphology of the middle and inner ear and endocast that are largely unknown in baenids. Semicircular canals of BDM 004 are virtually identical to those of Eubaena cephalica and consistent in dimensions to those of other turtle taxa, including anterior and posterior semicircular canals that are robust and taller than the common crus and diverge from each other at an angle of approximately 90°. The digital endocast reveals a moderately flexed brain with rounded cerebral hemispheres and minimal separation between the metencephalon and myelencephalon. Its well-preserved columella auris (stapes) is gracile with a posterodorsally flared basis columella. It arcs across the middle ear and flattens near its terminus. This study adds to the understanding of baenid middle and inner ear and neuroanatomical morphology and expands the morphological understanding of Pl. antiqua.

Quantitative Neuroanatomical Phenotyping of the Embryonic Mouse Brain.

Congenital neurodevelopmental anomalies are present from birth and are characterized by an abnormal development of one or more structures of the brain. Brain structural anomalies are highly comorbid with neurodevelopmental and neuropsychiatric disorders such as intellectual disability, autism spectrum disorders, epilepsy, and schizophrenia, and 80% are of genetic origin. We aim to address an important neurobiological question: How many genes regulate the normal anatomy of the brain during development. To do so, we developed a quantitative approach for the assessment of a total of 106 neuroanatomical parameters in mouse mutant embryos at embryonic day 18.5 across two planes commonly used in brain anatomical studies, the coronal and sagittal planes. In this article we describe the techniques we developed and explain why ultrastandardized procedures involving embryonic mouse brains are even more of prime importance for morphological phenotyping than adult mouse brains. We focus our analysis on brain size anomalies and on the most frequently altered brain regions including the cortex, corpus callosum, hippocampus, ventricles, caudate putamen, and cerebellum. Our protocols allow a standardized histology pipeline from embryonic mouse brain preparation to sectioning, staining, and scanning and neuroanatomical analyses at well-defined positions on the coronal and sagittal planes. Together, our protocols will help scientists in deciphering congenital neurodevelopmental anomalies and anatomical changes between groups of mouse embryos in health and genetic diseases. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Fixation and preparation of embryonic mouse brain samples Basic Protocol 2: Sectioning, staining, and scanning of embryonic mouse brain sections Basic Protocol 3: Coronal neuroanatomical measurements of embryonic mouse brain structures Basic Protocol 4: Sagittal neuroanatomical measurements of embryonic mouse brain structures.

Brain cutting and trimming.

The process of brain cutting followed by trimming is quite important to make adequate specimens for sufficient neuropathological observations. The protocol described herein is recommended as an optimized implementation for suitable preparation, which inevitably leads to an accurate neuropathological diagnosis. To obtain neuropathological cues, macroscopic observation of the brain before cutting presents an important opportunity. Gross examination provides a clue to the neuropathological diagnosis and shows clinicopathological correlations. Brain cutting should be preceded by a careful review of the clinical notes and consideration of the possible pathological diagnosis. Therefore, the medical staff associated with the patient should attend the procedure to provide clinical information. The process involves removing the brainstem and cerebellum from the cerebrum, sectioning the cerebrum, removing the cerebellum from the brainstem, and sectioning the cerebellum, brainstem and spinal cord followed by trimming. Trimming should be performed in accordance with the internationally accepted guidelines for the pathological diagnosis of different types of neurodegenerative diseases. In each stage acquiring clear photographs is significant, the observations must be concisely recorded, and which side of the specimen is to be sliced and stained has to be indicated. Additionally, it is necessary to photograph all trimmed tissues to assist with orientation of the brain in later assessments. The three-dimensional structure and individual differences have to be considered. These skills are essential, and knowledge of neuropathology, neurology and neuroanatomy is required for appropriately cutting and trimming of the brain.

Comparative anatomy of the encephalon of new world primates with emphasis for the Sapajus sp.

Studies about the anatomy of the New World Primates are scarce, mainly comparative neuroanatomy, then a morphological comparative analysis about the tropical Primates were performed and a effort was made for an Old World Primates and modern humans relationship for the obtained data; plus, comments about behavior e and allometry were performed to try link the high cognition and abilities of the Sapajus with the neuroanatomical results, however, despite the deep neuroanatomic data obtained, we do not found an intrinsic relation to explain that.

A historical perspective on training students to create standardized maps of novel brain structure: Newly-uncovered resonances between past and present research-based neuroanatomy curricula.

Recent efforts to reform postsecondary STEM education in the U.S. have resulted in the creation of course-based undergraduate research experiences (CUREs), which, among other outcomes, have successfully retained freshmen in their chosen STEM majors and provided them with a greater sense of identity as scientists by enabling them to experience how research is conducted in a laboratory setting. In 2014, we launched our own laboratory-based CURE, Brain Mapping & Connectomics (BMC). Now in its seventh year, BMC trains University of Texas at El Paso (UTEP) undergraduates to identify and label neuron populations in the rat brain, analyze their cytoarchitecture, and draw their detailed chemoarchitecture onto standardized rat brain atlas maps in stereotaxic space. Significantly, some BMC students produce atlas drawings derived from their coursework or from further independent study after the course that are being presented and/or published in the scientific literature. These maps should prove useful to neuroscientists seeking to experimentally target elusive neuron populations. Here, we review the procedures taught in BMC that have empowered students to learn about the scientific process. We contextualize our efforts with those similarly carried out over a century ago to reform U.S. medical education. Notably, we have uncovered historical records that highlight interesting resonances between our curriculum and that created at the Johns Hopkins University Medical School (JHUMS) in the 1890s. Although the two programs are over a century apart and were created for students of differing career levels, many aspects between them are strikingly similar, including the unique atlas-based brain mapping methods they encouraged students to learn. A notable example of these efforts was the brain atlas maps published by Florence Sabin, a JHUMS student who later became the first woman to be elected to the U.S. National Academy of Sciences. We conclude by discussing how the revitalization of century-old methods and their dissemination to the next generation of scientists in BMC not only provides student benefit and academic development, but also acts to preserve what are increasingly becoming "lost arts" critical for advancing neuroscience - brain histology, cytoarchitectonics, and atlas-based mapping of novel brain structure.

The nonhuman primate neuroimaging and neuroanatomy project.

Multi-modal neuroimaging projects such as the Human Connectome Project (HCP) and UK Biobank are advancing our understanding of human brain architecture, function, connectivity, and their variability across individuals using high-quality non-invasive data from many subjects. Such efforts depend upon the accuracy of non-invasive brain imaging measures. However, 'ground truth' validation of connectivity using invasive tracers is not feasible in humans. Studies using nonhuman primates (NHPs) enable comparisons between invasive and non-invasive measures, including exploration of how "functional connectivity" from fMRI and "tractographic connectivity" from diffusion MRI compare with long-distance connections measured using tract tracing. Our NonHuman Primate Neuroimaging & Neuroanatomy Project (NHP_NNP) is an international effort (6 laboratories in 5 countries) to: (i) acquire and analyze high-quality multi-modal brain imaging data of macaque and marmoset monkeys using protocols and methods adapted from the HCP; (ii) acquire quantitative invasive tract-tracing data for cortical and subcortical projections to cortical areas; and (iii) map the distributions of different brain cell types with immunocytochemical stains to better define brain areal boundaries. We are acquiring high-resolution structural, functional, and diffusion MRI data together with behavioral measures from over 100 individual macaques and marmosets in order to generate non-invasive measures of brain architecture such as myelin and cortical thickness maps, as well as functional and diffusion tractography-based connectomes. We are using classical and next-generation anatomical tracers to generate quantitative connectivity maps based on brain-wide counting of labeled cortical and subcortical neurons, providing ground truth measures of connectivity. Advanced statistical modeling techniques address the consistency of both kinds of data across individuals, allowing comparison of tracer-based and non-invasive MRI-based connectivity measures. We aim to develop improved cortical and subcortical areal atlases by combining histological and imaging methods. Finally, we are collecting genetic and sociality-associated behavioral data in all animals in an effort to understand how genetic variation shapes the connectome and behavior.

Neuroanatomy of expressive suppression: The role of the insula.

Expressive suppression is a response-focused regulatory strategy aimed at concealing the outward expression of emotion that is already underway. Expressive suppression requires the integration of interoception, proprioception, and social awareness to guide behavior in alignment with personal and interpersonal goals-all processes known to involve the insular cortex. Frontotemporal dementia (FTD) provides a useful patient model for studying the insula's role in socioemotional regulation. The insula is a key target of early atrophy in FTD, causing patients to lose the ability to represent the salience of internal and external conditions and to use these representations to guide behavior. We examined a sample of 59 patients with FTD, 52 patients with Alzheimer's disease (AD), and 38 neurologically healthy controls. Subjects viewed 2 disgust-eliciting films in the laboratory. During the first film, subjects were instructed to simply watch (emotional reactivity trial); during the second, they were instructed to hide their emotions (expressive suppression trial). Structural images from a subsample of participants (n = 42; 11 FTD patients, 11 AD patients, and 20 controls) were examined in conjunction with behavior. FreeSurfer was used to quantify regional gray matter volume in 41 empirically derived neural regions in both hemispheres. Of the 3 groups studied, FTD patients showed the least expressive suppression and had the smallest insula volumes, even after controlling for age, gender, and emotional reactivity. Among the brain regions examined, the insula was the only significant predictor of expressive suppression ability, with lower insula gray matter volume in both hemispheres predicting less expressive suppression. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Multi-aspect testing and ranking inference to quantify dimorphism in the cytoarchitecture of cerebellum of male, female and intersex individuals: a model applied to bovine brains.

The dimorphism among male, female and freemartin intersex bovines, focusing on the vermal lobules VIII and IX, was analyzed using a novel data analytics approach to quantify morphometric differences in the cytoarchitecture of digitalized sections of the cerebellum. This methodology consists of multivariate and multi-aspect testing for cytoarchitecture-ranking, based on neuronal cell complexity among populations defined by factors, such as sex, age or pathology. In this context, we computed a set of shape descriptors of the neural cell morphology, categorized them into three domains named size, regularity and density, respectively. The output and results of our methodology are multivariate in nature, allowing an in-depth analysis of the cytoarchitectonic organization and morphology of cells. Interestingly, the Purkinje neurons and the underlying granule cells revealed the same morphological pattern: female possessed larger, denser and more irregular neurons than males. In the Freemartin, Purkinje neurons showed an intermediate setting between males and females, while the granule cells were the largest, most regular and dense. This methodology could be a powerful instrument to carry out morphometric analysis providing robust bases for objective tissue screening, especially in the field of neurodegenerative pathologies.

Differences of neuroanatomy education for large and small numbers of students in China and Korea / Diferencias de educación en neuroanatomía de cantidades mayores y menores de estudiantes en China y Corea

In Southern Medical University, China, 1,200 medical students study neuroanatomy every year, whereas in Ajou University, Korea, only 45 medical students study neuroanatomy. The considerable difference of student numbers results in differences in educational situations. The purpose of this study was to investigate desirable neuroanatomy education methods for large and small numbers of students. The situations of neuroanatomy education in China and Korea were compared systematically. With a questionnaire survey, positive comments and recommendations for their counterparts were collected from the medical students (168 Chinese and 41 Koreans) and anatomists (6 Chinese and 3 Koreans). By reviewing the opinions, the Chinese and Korean anatomists could learn from each other to improve their strong points and make up for the weak points. The results also disclosed the common problems of neuroanatomy education, which could be relieved by developing the fitting book and the self-learning tools, such as lecture videos and stereoscopic computer models.

En la Universidad de Medicina del Sur, China, 1.200 estudiantes de medicina estudian la neuroanatomía cada año, mientras que en la Universidad de Ajou, Corea, solo 45 estudiantes de medicina estudian neuroanatomía. Esta considerable variable del número de estudiantes resulta en diferencias en las situaciones educativas. El propósito de este estudio fue investigar métodos de educación en neuroanatomía deseables para cantidades mayores y menores de estudiantes. Se compararon sistemáticamente las situaciones de educación en neuroanatomía en China y Corea. Por medio de una encuesta por cuestionario, se obtuvieron comentarios positivos y recomendaciones para sus contrapartes de los estudiantes de medicina (168 chinos y 41 coreanos) y anatomistas (6 chinos y 3 coreanos). Al revisar las opiniones, los anatomistas chinos y coreanos podrían aprender unos de otros para mejorar sus puntos de fortaleza y compensar los aspectos débiles. Los resultados también revelaron los problemas comunes de la educación en neuroanatomía, que podrían aliviarse desarrollando el libro de adaptación y las herramientas de autoaprendizaje, como videos de conferencias y modelos de computadora estereoscópica.

Sex chromosome aneuploidy alters the relationship between neuroanatomy and cognition.

Sex chromosome aneuploidy (SCA) increases the risk for cognitive deficits, and confers changes in regional cortical thickness (CT) and surface area (SA). Neuroanatomical correlates of inter-individual variation in cognitive ability have been described in health, but are not well-characterized in SCA. Here, we modeled relationships between general cognitive ability (estimated using full-scale IQ [FSIQ] from Wechsler scales) and regional estimates of SA and CT (from structural MRI scans) in both aneuploid (28 XXX, 55 XXY, 22 XYY, 19 XXYY) and typically-developing euploid (79 XX, 85 XY) individuals. Results indicated widespread decoupling of normative anatomical-cognitive relationships in SCA: we found five regions where SCA significantly altered SA-FSIQ relationships, and five regions where SCA significantly altered CT-FSIQ relationships. The majority of areas were characterized by the presence of positive anatomy-IQ relationships in health, but no or slightly negative anatomy-IQ relationships in SCA. Disrupted anatomical-cognitive relationships generalized from the full cohort to karyotypically defined subcohorts (i.e., XX-XXX; XY-XYY; XY-XXY), demonstrating continuity across multiple supernumerary SCA conditions. As the first direct evidence of altered regional neuroanatomical-cognitive relationships in supernumerary SCA, our findings shed light on potential genetic and structural correlates of the cognitive phenotype in SCA, and may have implications for other neurogenetic disorders.

Neuroanatomical underpinnings of autism symptomatology in carriers and non-carriers of the 22q11.2 microdeletion.

BACKGROUND: A crucial step to understanding the mechanistic underpinnings of autism spectrum disorder (ASD), is to examine if the biological underpinnings of ASD in genetic high-risk conditions, like 22q11.2 deletion syndrome (22q11.2DS), are similar to those in idiopathic illness. This study aimed to examine if ASD symptomatology in 22q11.2DS is underpinned by the same-or distinct-neural systems that mediate these symptoms in non-deletion carriers. METHODS: We examined vertex-wise estimates of cortical volume (CV), surface area (SA), and cortical thickness across 131 individuals between 6 and 25 years of age including (1) 50 individuals with 22q11.2DS, out of which n = 25 had a diagnosis of ASD, (2) 40 non-carriers of the microdeletion with a diagnosis of ASD (i.e., idiopathic ASD), and (3) 41 typically developing (TD) controls. We employed a 2-by-2 factorial design to identify neuroanatomical variability associated with the main effects of 22q11.2DS and ASD, as well as their interaction. Further, using canonical correlation analysis (CCA), we compared neuroanatomical variability associated with the complex (i.e., multivariate) clinical phenotype of ASD between 22q11.2 deletion carriers and non-carriers. RESULTS: The set of brain regions associated with the main effect of 22q11.2DS was distinct from the neuroanatomical underpinnings of the main effect of ASD. Moreover, significant 22q11.2DS-by-ASD interactions were observed for CV and SA in the dorsolateral prefrontal cortex, precentral gyrus, and posterior cingulate cortex, suggesting that the neuroanatomy of ASD is significantly modulated by 22q11.2DS (p < 0.01). We further established that the multivariate patterns of neuroanatomical variability associated with differences in symptom profiles significantly differed between 22q11.2 deletion carriers and non-carriers. LIMITATIONS: We employed a multicenter design to overcome single-site recruitment limitations; however, FreeSurfer-derived measures of surface anatomy have been shown to be highly reliable across scanner platforms and field strengths. Further, we controlled for gender to address the differing distribution between idiopathic ASD individuals and the other groups. Nonetheless, the gender distribution in our sample reflects that of the respective populations, adding to the generalizability of our results. Last, we included individuals with a relatively wide age range (i.e., 6-25 years). CONCLUSIONS: Our findings indicate that neuroanatomical correlates of ASD symptomatology in carriers of the 22q11.2 microdeletion diverge from those in idiopathic ASD.

The natverse, a versatile toolbox for combining and analysing neuroanatomical data.

To analyse neuron data at scale, neuroscientists expend substantial effort reading documentation, installing dependencies and moving between analysis and visualisation environments. To facilitate this, we have developed a suite of interoperable open-source R packages called the natverse. The natverse allows users to read local and remote data, perform popular analyses including visualisation and clustering and graph-theoretic analysis of neuronal branching. Unlike most tools, the natverse enables comparison across many neurons of morphology and connectivity after imaging or co-registration within a common template space. The natverse also enables transformations between different template spaces and imaging modalities. We demonstrate tools that integrate the vast majority of Drosophila neuroanatomical light microscopy and electron microscopy connectomic datasets. The natverse is an easy-to-use environment for neuroscientists to solve complex, large-scale analysis challenges as well as an open platform to create new code and packages to share with the community.

Basic Neuroanatomical Methods.

This unit covers some basic procedures that are common to a wide range of neuroanatomical protocols for brain tissue. Procedures are provided for preparation of unfixed fresh brain tissue as well as for perfusion fixation of animals to obtain fixed neural tissue. A variety of methods for sectioning are described, including frozen sectioning using a cryostat or microtome and sectioning with a vibratome. The choice of sectioning method depends on how the brain has been prepared and what histochemical method is to be used. A fluorescent immunohistochemical method to localize endogenous molecules as well as induced markers such as green fluorescent protein and red fluorescent protein is also provided. Additionally, three post-sectioning procedures are described: defatting of slide-mounted sections, fluorescent Nissl staining, and thionin staining of sections. Finally, support protocols are provided, describing a method for maintaining the correct order of cut tissue, whether rostral to caudal or lateral to medial; a procedure for subbing slides with gelatin, which is necessary in some protocols in order for sections to adhere to slides; and preparation of custom 3D-printed 10- or 20-well tissue plates and trays for subsequent immunostaining. Published 2019. U.S. Government. Basic Protocol 1: Preparation of unfixed fresh-frozen brain tissue Basic Protocol 2: Perfusion fixation Basic Protocol 3: Cryostat sectioning of frozen brain tissue Basic Protocol 4: Sliding-microtome sectioning of fixed brain tissue Basic Protocol 5: Vibratome and Compresstome sectioning Support Protocol 1: Tissue collection in a 1-in-10 series Support Protocol 2: Preparation of gelatin-subbed microscope slides Support Protocol 3: Custom 3D-printed 10- and 20-well tissue plates Basic Protocol 6: Post-sectioning procedures I: Fluorescent immunohistochemical localization Basic Protocol 7: Post-sectioning procedures II: Defatting Basic Protocol 8: Post-sectioning procedures III: Nissl staining Basic Protocol 9: Post-sectioning procedures IV: Thionin staining.

Generating brain matrices for zebra finch brain sectioning using three-dimensional printing technology.

BACKGROUND: The demand to sample brain regions in non-model species is increasing as more studies are integrating neurological data into behavioural, ecological or evolutionary analysis. However, the sampling operation is difficult for researchers without neuroscience background. It is also a challenge to collect neuroanatomical regions from animals in the field. NEW METHOD: Here we developed a new brain matrix for guiding researchers to section zebra finches' (Taeniopygia guttata) brains more steadily than by freehand trimming. Based on the 3D printing technology, we produced the zebra finch brain matrix from scratch. We also provided a step-by-step protocol to make brain matrices for any species with a brain size between that of shrews and dogs. RESULTS: The brain matrix could guide us to find the zebra finch's neuroanatomical landmarks, such as the hypothalamus, optic chiasm and occulomotor nerve. The matrix's channels near these landmarks could be used to section brains steadily and rapidly. COMPARISON WITH EXISTING METHODS: Standardized brain sectioning often requires expensive machines that may not be available in most laboratories or in the field, such as microtomes. In addition, machine-based trimming is time-consuming. Although commercial brain matrices can overcome these problems, they are only available for rats and mice. The brain matrices we developed are affordable to most laboratories and can be customised for non-model species in both lab and field experiments. CONCLUSIONS: The matrix-guided approach requires a relatively short training period and can allow researchers to properly and quickly sample brains, and thus will facilitate neuroscience-based interdisciplinary research.

Optimizing techniques for injecting DiI into the brain nuclei of neonatal mice P6 - C57Bl6/CBA.

DiI (1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate) is a lipophilic dye used to research the intracerebral connections of neonatal animals. This dye is most frequently used in the form of dry crystals, but injections of the marker in this state into the mouse brain nuclei, such as MHb (2 cases), LHbM (2 cases), LPA (3 cases) and TS (1 case), clearly revealed excessive lateral diffusion marker into the tissue. Eventually, all the investigated cases except the LPA gave a false positive result. To find the optimal method of marker injection,DiI-kerosene liquid films (3 cases in the MHb); DiI-kerosene gelatinous paste (3 cases in the MHb); injections of insoluble DiI crystals from a solution of 99% dmso (3 cases in the MHb) and 70% ethanol (3 cases in the MHb, 2 cases in the LHbM, 4 cases in the TS) were tested. Injections of DiI in the form of undissolved crystals from a 70% ethanol solution gave a minimal level of side diffusion in all the studied cases. This method of injection is optimal and recommended for use.

DNA sequencing in high-throughput neuroanatomy.

Mapping brain connectivity at single cell resolution is critical for understanding brain structure. For decades, such mapping has been principally approached with microscopy techniques, aiming to visualize neurons and their connections. However, these techniques are often very labor intensive and do not scale well to the complexity of mammalian brains. We recently leveraged the speed and parallelization of DNA sequencing to map the projections of thousands of single neurons in single experiments, and to map cortical mesoscale connectivity in single mice. Here, I review the state of sequencing-based neuroanatomy, and discuss future directions in synaptic connectivity mapping and comparative connectomics.

Evolution of neocortical folding: A phylogenetic comparative analysis of MRI from 34 primate species.

We conducted a comparative analysis of primate cerebral size and neocortical folding using magnetic resonance imaging data from 65 individuals belonging to 34 different species. We measured several neocortical folding parameters and studied their evolution using phylogenetic comparative methods. Our results suggest that the most likely model for neuroanatomical evolution is one where differences appear randomly (the Brownian Motion model), however, alternative models cannot be completely ruled out. We present estimations of the ancestral primate phenotypes as well as estimations of the rates of phenotypic change. Based on the Brownian Motion model, the common ancestor of primates may have had a folded cerebrum similar to that of a small lemur such as the aye-aye. Finally, we observed a non-linear relationship between fold wavelength and fold depth with cerebral volume. In particular, gyrencephalic primate neocortices across different groups exhibited a strikingly stable fold wavelength of about 12 mm (±20%), despite a 20-fold variation in cerebral volume. We discuss our results in the context of current theories of neocortical folding.