Neuroanatomical Studies Research Articles

FMRP regulates the subcellular distribution of cortical dendritic spine density in a non-cell-autonomous manner

Fragile X syndrome (FXS) is the most common form of intellectual disability that arises from the dysfunction of a single gene—Fmr1. The main neuroanatomical correlate of FXS is elevated dendritic spine density on cortical pyramidal neurons, which has been modeled in Fmr1-/Y mice. However, the cell-autonomous contribution of Fmr1 on cortical dendritic spine density has not been assessed. Even less is known about the role of Fmr1 in heterozygous female mosaic mice, which are a putative model for human Fmr1 full mutation carriers (i.e., are heterozygous for the full Fmr1-silencing mutation). In this neuroanatomical study, spine density in cortical pyramidal neurons of Fmr1+/− and Fmr1-/Y mice was studied at multiple subcellular compartments, layers, and brain regions. Spine density in Fmr1+/− mice is higher than WT but lower than Fmr1-/Y. Not all subcellular compartments in layer V Fmr1+/− and Fmr1-/Y cortical pyramidal neurons are equally affected: the apical dendrite, a key subcellular compartment, is principally affected over basal dendrites. Within apical dendrites, spine density is differentially affected across branch orders. Finally, identification of FMRP-positive and FMRP-negative neurons within Fmr1+/− permitted the study of the cell-autonomous effect of Fmr1 on spine density. Surprisingly, layer V cortical pyramidal spine density between FMRP-positive and FMRP-negative neurons does not differ, suggesting that the regulation of the primary neuroanatomical defect of FXS—elevated spine density—is non-cell-autonomous.

A neuroanatomical study of the feline brain using MRI and mulligan staining: functional and pathological considerations.

Despite multiple studies describing accurate diagnoses using advanced neuroimaging techniques, low and mid-field magnetic resonance imaging (MRI) are still the most frequent scanners in veterinary clinics. To date, these studies in cats do not show a clear distinction of nerve centres in MRI data. The objective of this study is to determine the efficacy of Mulligan histological staining as a tool in facilitating the location and identification of the main structures of the feline brain in MRI. This study aims to facilitate the interpretation of MRI obtained with these types of scanners. A total of 10 feline brains were used. One specimen was used for MRI (T2 sequence using a 1.5T scanner). The other 9 brains were sectioned and stained with the three Mulligan staining techniques (Mulligan, Le Masurier and Robert). The uptake of stain by the grey matter in these sections allowed the determination of the location and the limits of these nervous structures within the brain. The histological location of these structures was correlated with the MRI scans, leading to the successful identification of many small, indistinct nuclei. Mulligan staining is proposed as a tool that facilitates the location of nerve structures in comparison with data from the most frequently-used MRI scanners in veterinary clinics.

Common Challenges and Solutions Associated with the Preparation of Silicone-Injected Human Head and Neck Vessels for Anatomical Study.

Neuroanatomy laboratory training is crucial for the education of neurosurgery residents and medical students. Since the brain is a complex and three-dimensional structure, it is challenging to understand the anatomical relationship of the cortex, internal structures, arteries, and veins without appropriate adjuncts. Several injection agents—including the inks/dyes, latex, polyester, acrylic resins, phenol, polyethylene glycol, and phenoxyethanol—have been explored. Colored silicon injection protocols for the head and neck vessels’ perfusion have greatly aided the study of neuroanatomy and surgical planning. This report presents a colored silicone injection method in detail, and also highlights the technical shortcomings of the standard techniques and workarounds for common challenges during 35 human cadaveric head injections. The human cadaveric head preparation and the coloring of the head vessels are divided into decapitation, tissue fixation with 10% formalin, the placement of the Silastic tubing into the parent vessels, the cleaning of the vessels from clots, and the injection of the colored silicone into the vessels. We describe the technical details of the preparation, injection, and preservation of cadaveric heads, and outline common challenges during colored silicone injection, which include the dislocation of the Silastic tubing during the injection, the injection of the wrong or inappropriate colored silicone into the vessel, intracranial vessel perforation, the incomplete silicone casting of the vessel, and silicone leakage from small vessels in the neck. Solutions to these common challenges are provided. Ethyl alcohol fixed, colored human heads provided the long-term preservation of tissue, and improve the sample consistency and preservation for the teaching of neuroanatomy and surgical technique.

A Systematic Approach to Dissection of the Equine Brain-Evaluation of a Species-Adapted Protocol for Beginners and Experts.

Introduction of new imaging modalities for the equine brain have refocused attention on the horse as a natural model for ethological, neuroanatomical, and neuroscientific investigations. As opposed to imaging studies, strategies for equine neurodissection still lack a structured approach, standardization and reproducibility. In contrast to other species, where adapted protocols for sampling have been published, no comparable guideline is currently available for equids. Hence, we developed a species-specific slice protocol for whole brain vs. hemispheric dissection and tested its applicability and practicability in the field, as well as its neuroanatomical accuracy and reproducibility. Dissection steps are concisely described and depicted by schematic illustrations, photographs and instructional videos. Care was taken to show the brain in relation to the raters' hands, cutting devices and bench surface. Guidance is based on a minimum of external anatomical landmarks followed by geometric instructions that led to procurement of 14 targeted slabs. The protocol was performed on 55 formalin-fixed brains by three groups of investigators with different neuroanatomical skills. Validation of brain dissection outcomes addressed the aptitude of slabs for neuroanatomical studies as opposed to simplified routine diagnostic purposes. Across all raters, as much as 95.2% of slabs were appropriate for neuroanatomical studies, and 100% of slabs qualified for a routine diagnostic setting. Neither autolysis nor subfixation significantly affected neuroanatomical accuracy score, while a significant negative effect was observed with brain extraction artifacts. Procedure times ranged from 14 to 66 min and reached a mean duration of 23.25 ± 7.93 min in the last of five trials in inexperienced raters vs. 16 ± 2.83 min in experts, while acceleration of the dissection did not negatively impact neuroanatomical accuracy. This protocol, derived analogously to the consensus report of the International Veterinary Epilepsy Task Force in dogs and cats, allows for systematic, quick and easy dissection of the equine brain, even for inexperienced investigators. Obtained slabs feature virtually all functional subcompartments at suitable planes for both diagnostic and neuroscientific investigations and complement the data obtained from imaging studies. The instructive protocol and brain dissection videos are available in Supplementary Material.

An Angiotensin-Responsive Connection from the Lamina Terminalis to the Paraventricular Nucleus of the Hypothalamus Evokes Vasopressin Secretion to Increase Blood Pressure in Mice.

Blood pressure is controlled by endocrine, autonomic, and behavioral responses that maintain blood volume and perfusion pressure at levels optimal for survival. Although it is clear that central angiotensin type 1a receptors (AT1aR; encoded by the Agtr1a gene) influence these processes, the neuronal circuits mediating these effects are incompletely understood. The present studies characterize the structure and function of AT1aR neurons in the lamina terminalis (containing the median preoptic nucleus and organum vasculosum of the lamina terminalis), thereby evaluating their roles in blood pressure control. Using male Agtr1a-Cre mice, neuroanatomical studies reveal that AT1aR neurons in the area are largely glutamatergic and send projections to the paraventricular nucleus of the hypothalamus (PVN) that appear to synapse onto vasopressin-synthesizing neurons. To evaluate the functionality of these lamina terminalis AT1aR neurons, we virally delivered light-sensitive opsins and then optogenetically excited or inhibited the neurons while evaluating cardiovascular parameters or fluid intake. Optogenetic excitation robustly elevated blood pressure, water intake, and sodium intake, while optogenetic inhibition produced the opposite effects. Intriguingly, optogenetic excitation of these AT1aR neurons of the lamina terminalis also resulted in Fos induction in vasopressin neurons within the PVN and supraoptic nucleus. Further, within the PVN, selective optogenetic stimulation of afferents that arise from these lamina terminalis AT1aR neurons induced glutamate release onto magnocellular neurons and was sufficient to increase blood pressure. These cardiovascular effects were attenuated by systemic pretreatment with a vasopressin-1a-receptor antagonist. Collectively, these data indicate that excitation of lamina terminalis AT1aR neurons induces neuroendocrine and behavioral responses that increase blood pressure.SIGNIFICANCE STATEMENT Hypertension is a widespread health problem and risk factor for cardiovascular disease. Although treatments exist, a substantial percentage of patients suffer from "drug-resistant" hypertension, a condition associated with increased activation of brain angiotensin receptors, enhanced sympathetic nervous system activity, and elevated vasopressin levels. The present study highlights a role for angiotensin Type 1a receptor expressing neurons located within the lamina terminalis in regulating endocrine and behavioral responses that are involved in maintaining cardiovascular homeostasis. More specifically, data presented here reveal functional excitatory connections between angiotensin-sensitive neurons in the lamina terminals and vasopressin neurons in the paraventricular nucleus of the hypothalamus, and further indicate that activation of this circuit raises blood pressure. These neurons may be a promising target for antihypertensive therapeutics.

P.392 Cerebellar influence on the thalamo-prefrontal firing patterns during fear extinction

Based on its diverse and wide-spread patterns of connectivity, primate posteromedial cortex (PMC) is well positioned to support roles in several aspects of sensory-, cognitive- and motor-related processing. Previous work in both humans and non-human primates (NHPs) using resting-state functional MRI (rs-fMRI) suggests that a subregion of PMC, the medial parieto-occipital cortex (mPOC), by virtue of its intrinsic functional connectivity (FC) with visual cortex, may only play a role in higher-order visual processing. Recent neuroanatomical tracer studies in NHPs, however, demonstrate that mPOC also has prominent cortico-cortical connections with several frontoparietal structures involved in movement planning and control, a finding consistent with increasing observations of reach- and grasp-related activity in the mPOC of both NHPs and humans. To reconcile these observations, here we used rs-fMRI data collected from both awake humans and anesthetized macaque monkeys to more closely examine and compare parcellations of mPOC across species and explore the FC patterns associated with these subdivisions. Seed-based and voxel-wise hierarchical cluster analyses revealed four broad spatially separated functional boundaries that correspond with graded differences in whole-brain FC patterns in each species. The patterns of FC observed are consistent with mPOC forming a critical hub of networks involved in action planning and control, spatial navigation, and working memory. In addition, our comparison between species indicates that while there are several similarities, there may be some species-specific differences in functional neural organization. These findings and the associated theoretical implications are discussed.

Hindbrain and Spinal Cord Contributions to the Cutaneous Sensory Innervation of the Larval Zebrafish Pectoral Fin.

Vertebrate forelimbs contain arrays of sensory neuron fibers that transmit signals from the skin to the nervous system. We used the genetic toolkit and optical clarity of the larval zebrafish to conduct a live imaging study of the sensory neurons innervating the pectoral fin skin. Sensory neurons in both the hindbrain and the spinal cord innervate the fin, with most cells located in the hindbrain. The hindbrain somas are located in rhombomere seven/eight, laterally and dorsally displaced from the pectoral fin motor pool. The spinal cord somas are located in the most anterior part of the cord, aligned with myomere four. Single cell reconstructions were used to map afferent processes and compare the distributions of processes to soma locations. Reconstructions indicate that this sensory system breaks from the canonical somatotopic organization of sensory systems by lacking a clear organization with reference to fin region. Arborizations from a single cell branch widely over the skin, innervating the axial skin, lateral fin surface, and medial fin surface. The extensive branching over the fin and the surrounding axial surface suggests that these fin sensory neurons report on general conditions of the fin area rather than providing fine location specificity, as has been demonstrated in other vertebrate limbs. With neuron reconstructions that span the full primary afferent arborization from the soma to the peripheral cutaneous innervation, this neuroanatomical study describes a system of primary sensory neurons and lays the groundwork for future functional studies.

Comparative neuroanatomical study of the amygdala and fear conditioning in Nigerian breeds of Artiodactyla: Sheep (Uda) and goats (Red Sokoto)

The aim of this study was to evaluate fear condition responses in sheep and goat and to relate this to the neuroarchitecture of their amygdala. Forty adult sheep (Uda breed) and 40 adult goats (Red Sokoto breed) were fear-conditioned by associating the sound of a car horn (neutral stimuli) with water spray (aversive stimuli) and the fear response was determined by direct observation of the behavior of the sheep and goats and measuring their flight distances and escape time. Eight groups were studied, each comprising of 10 animals (five sheep and five goats). Goats and sheep were tested alternately in the morning of every day of the week for three consecutive weeks, in which 4 days was used for habituation and 3 days for testing. Histologically, neurons in the central and basolateral complex of the amygdala were studied and analyzed using Nissl and golgi staines. Behaviorally, goats elicited an active avoidance response expressed as flight with concomitant intense flight distances (p < .001) compared to sheep. Although, sheep had larger brain parameters, it showed attenuated basolateral amygdala cytoarchitecture consistent with reduced fear perception and response. Goats had significantly more densely distributed pyramidal and spiny stellate neurons in the basolateral amygdala while sheep showed more non-pyramidal and aspiny neurons. These results provide interesting practical perspectives on how adaptions in the amygdala coincides with alterations in fear conditioning in domestic animals and may be the basis for the higher incidence of the sheep in automobile accidents than goats in developing countries especially Africa.

Continuous block at the proximal end of the adductor canal provides better analgesia compared to that at the middle of the canal after total knee arthroplasty: a randomized, double-blind, controlled trial

BackgroundThe optimal position for continuous adductor canal block (ACB) for analgesia after total knee anthroplasty (TKA) remians controversial, mainly due to high variability in the localization of the the adductor canal (AC). Latest neuroanatomy studies show that the nerve to vastus medialis plays an important role in innervating the anteromedial aspect of the knee and dives outside of the exact AC at the proximal end of the AC. Therefore, we hypothesized that continuous ACB at the proximal end of the exact AC could provide a better analgesic effect after TKA compared with that at the middle of the AC (which appeared to only block the saphenous nerve).MethodsSixty-two adult patients who were scheduled for a unilateral TKA were randomized to receive continuous ACB at the proximal end or middle of the AC. All patients received patient-controlled intravenous analgesia with sufentanil postoperatively. The primary outcome measure was cumulative sufentanil consumption within 24 h after the surgery, which was analyzed using Mann-Whitney U tests. P-values < 0.05 (two-sided) were considered statistically significant. The secondary outcomes included postoperative sufentanil consumption at other time points, pain at rest and during passive knee flexion, quadriceps motor strength, and other recovery related paramaters.ResultsSixty patients eventually completed the study (30/group). The 24-h sufentanil consumption was 0.22 μg/kg (interquartile range [IQR]: 0.15–0.40 μg/kg) and 0.39 μg/kg (IQR: 0.23–0.52 μg/kg) in the proximal end and middle groups (P = 0.026), respectively. There were no significant inter-group differences in sufentanil consumption at other time points, pain at rest and during passive knee flexion, quadriceps motor strength, and other recovery related paramaters.ConclusionsContinuous ACB at the proximal end of the AC has a better opioid-sparing effect without a significant influence on quadriceps motor strength compared to that at the middle of the AC after TKA. These findings indicates that a true ACB may not produce the effective analgesia, instead, the proximal end AC might be a more suitable block to alleviate pain after TKA.Trial registrationThis study was registered at ClinicalTrials.gov (NCT03942133; registration date: May 06, 2019; enrollment date: May 11, 2019).

Functional parcellation of the hippocampus by semi-supervised clustering of resting state fMRI data

Many unsupervised methods are widely used for parcellating the brain. However, unsupervised methods aren’t able to integrate prior information, obtained from such as exiting functional neuroanatomy studies, to parcellate the brain, whereas the prior information guided semi-supervised method can generate more reliable brain parcellation. In this study, we propose a novel semi-supervised clustering method for parcellating the brain into spatially and functionally consistent parcels based on resting state functional magnetic resonance imaging (fMRI) data. Particularly, the prior supervised and spatial information is integrated into spectral clustering to achieve reliable brain parcellation. The proposed method has been validated in the hippocampus parcellation based on resting state fMRI data of 20 healthy adult subjects. The experimental results have demonstrated that the proposed method could successfully parcellate the hippocampus into head, body and tail parcels. The distinctive functional connectivity patterns of these parcels have further demonstrated the validity of the parcellation results. The effects of aging on the three hippocampus parcels’ functional connectivity were also explored across the healthy adult subjects. Compared with state-of-the-art methods, the proposed method had better performance on functional homogeneity. Furthermore, the proposed method had good test–retest reproducibility validated by parcellating the hippocampus based on three repeated resting state fMRI scans from 24 healthy adult subjects.

Genicular Nerve Radiofrequency Ablation for Chronic Knee Pain Using a Three-Tined Electrode: A Technical Description and Case Series.

Genicular nerve radiofrequency ablation (RFA) for the treatment of chronic knee pain has traditionally targeted the superomedial, superolateral, and inferomedial genicular nerves. However, recent cadaveric studies of knee neuroanatomy demonstrate varied locations of these specific nerves as well as additional articular nerves. This work suggests that traditional genicular nerve RFA lesion locations may be inadequate. 1) To describe a novel protocol utilizing a three-tined RFA electrode to target the superomedial (SMGN), superolateral (SLGN), and inferomedial genicular nerves (IMGN), as well as the terminal articular branches of the nerves to the vastus medialis (NVM), intermedius (NVI), and lateralis (NVL). 2) To assess the ability of this technique to reduce chronic knee pain. Case series of consecutive patients with six or more months of refractory knee pain who underwent genicular nerve RFA according to the novel protocol described. Seven discrete RFA lesions were placed to target the SMGN, NVM, NVI, NVL, SLGN, and IGMN. Eleven patients underwent RFA, nine with knee osteoarthritis and two postarthroplasty. At one month, 91% (95% CI = 59-100%), 82% (95% CI = 48-98%), and 9% (95% CI = 2-41%), of patients reported ≥50%, ≥80%, and 100% improvement in knee pain on the numeric rating scale, respectively. These results were sustained at six months. There were no complications. These preliminary data suggest the feasibility and possible effectiveness of genicular nerve RFA using the described novel protocol including a three-tined electrode. Larger-scale studies with comparative groups are warranted.

Chemical Sectioning Fluorescence Tomography: High-Throughput, High-Contrast, Multicolor, Whole-Brain Imaging at Subcellular Resolution

A thorough neuroanatomical study of the brain architecture is crucial for our understanding of its cellular compositions, connections and working mechanisms. However, the fine and multiscale features of neuron structures make it challenging for the microscopic imaging, as one cannot have both high-contrast, and high-throughput at the same time in such imaging. Here, we proposed the concept of chemical sectioning fluorescence tomography (CSFT) to solve this problem. Based on the switching characteristics of the fluorescent proteins (FPs), we introduced the chemical sectioning (CS) effect into the block-surface microscopy, turn off all FP molecules by chemical methods, and only light those in the top section as thin as submicron for imaging, so as to achieve high-throughput, high-contrast imaging without background. We developed a three-color wide-field fluorescence micro-optical sectioning tomography (fMOST) system. We proved that CSFT method was compatible with resin-embedded multicolor FPs and could be feasibly used in whole-brain imaging on diverse genetically or virally labeled long-projecting neurons and interneurons to the systematic reconstruction degree. We demonstrated mouse whole-brain imaging at the subcellular resolution, as well as the power for quantitative acquisition of synaptic connection-related pyramidal dendritic spines and axon boutons on the brain-wide scale.at the complete single-neuron level. We believe that CSFT method would greatly facilitate our understanding of the brain-wide neuron networks.

Spontaneous age-related changes of attention in unmedicated boys with attention deficit hyperactivity disorder

Objective Neuropsychological, neuroanatomical, and electrophysiological studies have reported a steady increase in the different attention types until the age of 10 years. Moreover, differences between healthy control (HC) boys and those with attention deficit hyperactivity disorder (ADHD) become nonsignificant in late childhood. This cross-sectional study aimed to perform a comparative analysis of attentional processing in boys with ADHD and HC in the 6:00-10:11 years age range. Methods: Age-related changes in attentional processing were compared between Caucasian Turkic boys (72-131 months of age) with ADHD (n = 144) and HC (n = 112). Selective, focused, and inhibitory attention were measured using the Stroop Test (5 scores); sustained attention was measured using the Cancellation Test (3 scores); and attention span was measured using the Visual Aural Digit Span Test-Revised (6 scores). Results: At the age of 6 years, the ADHD group had a significantly lower performance for all attention types. By the age of 10 years, there were no significant between-group differences. However, the component structure of the neuropsychological test scores in the ADHD group differed from that in the HC group and previous studies. Conclusions: Attentional processing in boys with ADHD changes within the age-range of 6:00-10:00 years where it finally becomes similar to that in HC boys. This delayed maturation is consistent with the maturational lag model of ADHD. However, there was a between-group difference in the component structure of attentional processing, which is consistent with the maturational deviance model of ADHD.

The effects of early-treated phenylketonuria on volumetric measures of the cerebellum

Past murine studies of phenylketonuria (PKU) have documented significant effects on cerebellum at both the gross and cellular levels. The profile of neurocognitive and motor difficulties associated with early-treated PKU (ETPKU) is also consistent with potential cerebellar involvement. Previous neuroanatomical studies of cerebellum in patients with PKU, however, have yielded mixed results. The objective of the present study was to further examine potential differences in cerebellar morphometry between individuals with and without ETPKU. To this end, we analyzed high resolution T1-weighted MR images from a sample of 20 individuals with ETPKU and an age-matched comparison group of 20 healthy individuals without PKU. Measurements of whole brain volume, whole cerebellum volume, cerebellar gray matter volume, and cerebellar white matter volume were collected by means of semiautomatic volumetric analysis. Data analysis revealed no significant group differences in whole brain volume, whole cerebellar volume, or cerebellar white matter volume. A significant reduction in cerebellar gray matter volume, however, was observed for the ETPKU group compared to the non-PKU comparison group. These findings expand on previous animal work suggesting that cerebellar gray matter is impacted by PKU. It is also consistent with the hypothesis that the cognitive difficulties experienced by individuals with ETPKU may be related to disruptions in gray matter. Additional studies are needed to fully elucidate the timing and extent of the impact of ETPKU on cerebellum and the associated neurocognitive consequences.

Effects of MP2RAGE B1+ sensitivity on inter-site T1 reproducibility and hippocampal morphometry at 7T

Most neuroanatomical studies are based on T1-weighted MR images, whose intensity profiles are not solely determined by the tissue's longitudinal relaxation times (T1), but also affected by varying non-T1 contributions, hampering data reproducibility. In contrast, quantitative imaging using the MP2RAGE sequence, for example, allows direct characterization of the brain based on the tissue property of interest. Combined with 7 Tesla (7T) MRI, this offers unique opportunities to obtain robust high-resolution brain data characterized by a high reproducibility, sensitivity and specificity. However, specific MP2RAGE parameter choices – e.g., to emphasize intracortical myelin-dependent contrast variations – can substantially impact image quality and cortical analyses through remnants of B1+-related intensity variations, as illustrated in our previous work. To follow up on this: we (1) validate this protocol effect using a dataset acquired with a particularly B1+ insensitive set of MP2RAGE parameters combined with parallel transmission excitation; and (2) extend our analyses to evaluate the effects on hippocampal morphometry. The latter remained unexplored initially, but can provide important insights related to generalizability and reproducibility of neurodegenerative research using 7T MRI. We confirm that B1+ inhomogeneities have a considerably variable effect on cortical T1 estimates, as well as on hippocampal morphometry depending on the MP2RAGE setup. While T1 differed substantially across datasets initially, we show the inter-site T1 comparability improves after correcting for the spatially varying B1+ field using a separately acquired Sa2RAGE B1+ map. Finally, removal of B1+ residuals affects hippocampal volumetry and boundary definitions, particularly near structures characterized by strong intensity changes (e.g. cerebral spinal fluid). Taken together, we show that the choice of MP2RAGE parameters can impact T1 comparability across sites and present evidence that hippocampal segmentation results are modulated by B1+ inhomogeneities. This calls for careful (1) consideration of sequence parameters when setting acquisition protocols, as well as (2) acquisition of a B1+ map to correct MP2RAGE data for potential B1+ variations to allow comparison across datasets.

Cyto- and myeloarchitectural brain atlas of the pale spear-nosed bat (Phyllostomus discolor) in CT Aided Stereotaxic Coordinates

The pale spear-nosed bat Phyllostomus discolor, a microchiropteran bat, is well established as an animal model for research on the auditory system, echolocation and social communication of species-specific vocalizations. We have created a brain atlas of Phyllostomus discolor that provides high-quality histological material for identification of brain structures in reliable stereotaxic coordinates to strengthen neurobiological studies of this key species. The new atlas combines high-resolution images of frontal sections alternately stained for cell bodies (Nissl) and myelinated fibers (Gallyas) at 49 rostrocaudal levels, at intervals of 350 µm. To facilitate comparisons with other species, brain structures were named according to the widely accepted Paxinos nomenclature and previous neuroanatomical studies of other bat species. Outlines of auditory cortical fields, as defined in earlier studies, were mapped onto atlas sections and onto the brain surface, together with the architectonic subdivisions of the neocortex. X-ray computerized tomography (CT) of the bat’s head was used to establish the relationship between coordinates of brain structures and the skull. We used profile lines and the occipital crest as skull landmarks to line up skull and brain in standard atlas coordinates. An easily reproducible protocol allows sectioning of experimental brains in the standard frontal plane of the atlas. An electronic version of the atlas plates and supplementary material is available from https://doi.org/10.12751/g-node.8bbcxy

Volumetric analysis and morphological assessment of the ascending olfactory pathway in an elasmobranch and a teleost using diceCT.

The size (volume or mass) of the olfactory bulbs in relation to the whole brain has been used as a neuroanatomical proxy for olfactory capability in a range of vertebrates, including fishes. Here, we use diffusible iodine-based contrast-enhanced computed tomography (diceCT) to test the value of this novel bioimaging technique for generating accurate measurements of the relative volume of the main olfactory brain areas (olfactory bulbs, peduncles, and telencephalon) and to describe the morphological organisation of the ascending olfactory pathway in model fish species from two taxa, the brownbanded bamboo shark Chiloscyllium punctatum and the common goldfish Carassius auratus. We also describe the arrangement of primary projections to the olfactory bulb and secondary projections to the telencephalon in both species. Our results identified substantially larger olfactory bulbs and telencephalon in C. punctatum compared to C. auratus (comprising approximately 5.2% vs. 1.8%, and 51.8% vs. 11.8% of the total brain volume, respectively), reflecting differences between taxa, but also possibly in the role of olfaction in the sensory ecology of these species. We identified segregated primary projections to the bulbs, associated with a compartmentalised olfactory bulb in C. punctatum, which supports previous findings in elasmobranch fishes. DiceCT imaging has been crucial for visualising differences in the morphological organisation of the olfactory system of both model species. We consider comparative neuroanatomical studies between representative species of both elasmobranch and teleost fish groups are fundamental to further our understanding of the evolution of the olfactory system in early vertebrates and the neural basis of olfactory abilities.

Anterograde Neuronal Circuit Tracers Derived from Herpes Simplex Virus 1: Development, Application, and Perspectives.

Herpes simplex virus type 1 (HSV-1) has great potential to be applied as a viral tool for gene delivery or oncolysis. The broad infection tropism of HSV-1 makes it a suitable tool for targeting many different cell types, and its 150 kb double-stranded DNA genome provides great capacity for exogenous genes. Moreover, the features of neuron infection and neuron-to-neuron spread also offer special value to neuroscience. HSV-1 strain H129, with its predominant anterograde transneuronal transmission, represents one of the most promising anterograde neuronal circuit tracers to map output neuronal pathways. Decades of development have greatly expanded the H129-derived anterograde tracing toolbox, including polysynaptic and monosynaptic tracers with various fluorescent protein labeling. These tracers have been applied to neuroanatomical studies, and have contributed to revealing multiple important neuronal circuits. However, current H129-derived tracers retain intrinsic drawbacks that limit their broad application, such as yet-to-be improved labeling intensity, potential nonspecific retrograde labeling, and high toxicity. The biological complexity of HSV-1 and its insufficiently characterized virological properties have caused difficulties in its improvement and optimization as a viral tool. In this review, we focus on the current H129-derived viral tracers and highlight strategies in which future technological development can advance its use as a tool.

Measurements of neuronal morphological variation across the rat neocortex

Neuron morphology is highly variable across the mammalian brain. It is thought that these attributes of neuronal cell shape, such as soma surface area and branching frequency, are determined by biological function and information processing. In this study, a large data set of neurons across the rat neocortex were clustered by their anatomical characters for evidence of distinctiveness among neocortical regions and the somatosensory layers. This data set of neuronal morphologies was compiled from 31 different lab sources with a validation procedure so that data records are potentially comparable across research studies. With this large set of heterogeneous data and by clustering analysis, this study shows that neuronal morphological traits overlap among neocortical and somatosensory regions. In the context of past neuroanatomical studies, this result is not congruent with tissue level analysis and strongly suggests further sampling of neuronal data to lessen the effect of confounding factors, such as the influence of different methodologies from use of heterogeneous samples of neuronal data.

Dorsal prefrontal and premotor cortex of the ferret as defined by distinctive patterns of thalamo-cortical projections

Recent studies of the neurobiology of the dorsal frontal cortex (FC) of the ferret have illuminated its key role in the attention network, top-down cognitive control of sensory processing, and goal directed behavior. To elucidate the neuroanatomical regions of the dorsal FC, and delineate the boundary between premotor cortex (PMC) and dorsal prefrontal cortex (dPFC), we placed retrograde tracers in adult ferret dorsal FC anterior to primary motor cortex and analyzed thalamo-cortical connectivity. Cyto- and myeloarchitectural differences across dorsal FC and the distinctive projection patterns from thalamic nuclei, especially from the subnuclei of the medial dorsal (MD) nucleus and the ventral thalamic nuclear group, make it possible to clearly differentiate three separate dorsal FC fields anterior to primary motor cortex: polar dPFC (dPFCpol), dPFC, and PMC. Based on the thalamic connectivity, there is a striking similarity of the ferret’s dorsal FC fields with other species. This possible homology opens up new questions for future comparative neuroanatomical and functional studies.