Sulcal Patterns Research Articles

Molecular genetics, neuroimaging outcomes, and structural analyses of novel and recurrent variants of WDR62 gene in two consanguineous Pakistani families with autosomal recessive primary microcephaly.

Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental and genetically heterogeneous disorder, characterized by small cranium size (> -3 SD below mean) and often results in varying degree of intellectual disability. Thirty genes have been identified for the etiology of this disorder due to its clinical and genetic heterogeneity. Here, we report two consanguineous Pakistani families affected with MCPH exhibiting mutation in WDR62 gene. The investigation approach involved Next Generation Sequencing (NGS) gene panel sequencing coupled with linkage analysis followed by validation of identified variants through automated Sanger sequencing and Barcode-Tagged (BT) sequencing. The molecular genetic analysis revealed one novel splice site variant (NM_001083961.2(WDR62):c.1372-1del) in Family A and one known exonic variant NM_001083961.2(WDR62):c.3936dup (p.Val1313Argfs*18) in Family B. Magnetic Resonance Imaging (MRI) scans were also employed to gain insights into the structural architecture of affected individuals. Neurological assessments showed the reduced gyral and sulcal patterns along with normal corpus callosum in affected individuals harboring novel variant. In silico assessments of the identified variants were conducted using different tools to confirm the pathogenicity of these variants. Through In silico analyses, both variants were identified as disease causing and protein modeling of exonic variant indicates subtle conformational alterations in prophesied protein structure. This study identifies a novel variant (c.1372-1del) and a recurrent pathogenic variant c.3936dup (p.Val1313Argfs*18) in the WDR62 gene among the Pakistani population, expanding the mutation spectrum for MCPH. These findings emphasize the importance of genetic counseling and awareness to reduce consanguinity and address the burden of this disorder.

Meta-regression of sulcal patterns, clinical and environmental factors on neurodevelopmental outcomes in participants with multiple CHD types.

Congenital heart disease affects 1% of infants and is associated with impaired neurodevelopment. Right- or left-sided sulcal features correlate with executive function among people with Tetralogy of Fallot or single ventricle congenital heart disease. Studies of multiple congenital heart disease types are needed to understand regional differences. Further, sulcal pattern has not been studied in people with d-transposition of the great arteries. Therefore, we assessed the relationship between sulcal pattern and executive function, general memory, and processing speed in a meta-regression of 247 participants with three congenital heart disease types (114 single ventricle, 92 d-transposition of the great arteries, and 41 Tetralogy of Fallot) and 94 participants without congenital heart disease. Higher right hemisphere sulcal pattern similarity was associated with improved executive function (Pearson r = 0.19, false discovery rate-adjusted P = 0.005), general memory (r = 0.15, false discovery rate P = 0.02), and processing speed (r = 0.17, false discovery rate P = 0.01) scores. These positive associations remained significant in for the d-transposition of the great arteries and Tetralogy of Fallot cohorts only in multivariable linear regression (estimated change β = 0.7, false discovery rate P = 0.004; β = 4.1, false discovery rate P = 0.03; and β = 5.4, false discovery rate P = 0.003, respectively). Duration of deep hypothermic circulatory arrest was also associated with outcomes in the multivariate model and regression tree analysis. This suggests that sulcal pattern may provide an early biomarker for prediction of later neurocognitive challenges among people with congenital heart disease.

The role of cortical structural variance in deep learning-based prediction of fetal brain age.

Deep-learning-based brain age estimation using magnetic resonance imaging data has been proposed to identify abnormalities in brain development and the risk of adverse developmental outcomes in the fetal brain. Although saliency and attention activation maps have been used to understand the contribution of different brain regions in determining brain age, there has been no attempt to explain the influence of shape-related cortical structural features on the variance of predicted fetal brain age. We examined the association between the predicted brain age difference (PAD: predicted brain age-chronological age) from our convolution neural networks-based model and global and regional cortical structural measures, such as cortical volume, surface area, curvature, gyrification index, and folding depth, using regression analysis. Our results showed that global brain volume and surface area were positively correlated with PAD. Additionally, higher cortical surface curvature and folding depth led to a significant increase in PAD in specific regions, including the perisylvian areas, where dramatic agerelated changes in folding structures were observed in the late second trimester. Furthermore, PAD decreased with disorganized sulcal area patterns, suggesting that the interrelated arrangement and areal patterning of the sulcal folds also significantly affected the prediction of fetal brain age. These results allow us to better understand the variance in deep learning-based fetal brain age and provide insight into the mechanism of the fetal brain age prediction model.

Machine learning in small sample neuroimaging studies: Novel measures for schizophrenia analysis.

Novel features derived from imaging and artificial intelligence systems are commonly coupled to construct computer-aided diagnosis (CAD) systems that are intended as clinical support tools or for investigation of complex biological patterns. This study used sulcal patterns from structural images of the brain as the basis for classifying patients with schizophrenia from unaffected controls. Statistical, machine learning and deep learning techniques were sequentially applied as a demonstration of how a CAD system might be comprehensively evaluated in the absence of prior empirical work or extant literature to guide development, and the availability of only small sample datasets. Sulcal features of the entire cerebral cortex were derived from 58 schizophrenia patients and 56 healthy controls. No similar CAD systems has been reported that uses sulcal features from the entire cortex. We considered all the stages in a CAD system workflow: preprocessing, feature selection and extraction, and classification. The explainable AI techniques Local Interpretable Model-agnostic Explanations and SHapley Additive exPlanations were applied to detect the relevance of features to classification. At each stage, alternatives were compared in terms of their performance in the context of a small sample. Differentiating sulcal patterns were located in temporal and precentral areas, as well as the collateral fissure. We also verified the benefits of applying dimensionality reduction techniques and validation methods, such as resubstitution with upper bound correction, to optimize performance.

Are numerical abilities determined at early age? A brain morphology study in children and adolescents with and without developmental dyscalculia

The intraparietal sulcus (IPS) has been associated with numerical processing. A recent study reported that the IPS sulcal pattern was associated with arithmetic and symbolic number abilities in children and adults. In the present study, we evaluated the link between numerical abilities and the IPS sulcal pattern in children with Developmental Dyscalculia (DD) and typically developing children (TD), extending previous analyses considering other sulcal features and the postcentral sulcus (PoCS). First, we confirm the longitudinal sulcal pattern stability of the IPS and the PoCS. Second, we found a lower proportion of left sectioned IPS and a higher proportion of a double-horizontal IPS shape bilaterally in DD compared to TD. Third, our analyses revealed that arithmetic is the only aspect of numerical processing that is significantly related to the IPS sulcal pattern (sectioned vs not sectioned), and that this relationship is specific to the left hemisphere. And last, correlation analyses of age and arithmetic in children without a sectioned left IPS indicate that although they may have an inherent disadvantage in numerical abilities, these may improve with age. Thus, our results indicate that only the left IPS sulcal pattern is related to numerical abilities and that other factors co-determine numerical abilities.

An MRI Study of Morphology, Asymmetry, and Sex Differences of Inferior Precentral Sulcus

Numerous studies utilizing magnetic resonance imaging (MRI) have observed sex and interhemispheric disparities in sulcal morphology, which could potentially underpin certain functional disparities in the human brain. Most of the existing research examines the precentral sulcus comprehensively, with a rare focus on its subsections. To explore the morphology, asymmetry, and sex disparities within the inferior precentral sulcus (IPCS), we acquired 3.0T magnetic resonance images from 92 right-handed Chinese adolescents. Brainvisa was used to reconstruct the IPCS structure and calculate its mean depth (MD). Based on the morphological patterns of IPCS, it was categorized into five distinct types. Additionally, we analyzed four different types of spatial relationships between IPCS and inferior frontal sulcus (IFS). There was a statistically significant sex disparity in the MD of IPCS, primarily observed in the right hemisphere. Females exhibited significantly greater asymmetry in the MD of IPCS compared to males. No statistically significant sex or hemispheric variations were identified in sulcal patterns. Our findings expand the comprehension of inconsistencies in sulcal structure, while also delivering an anatomical foundation for the study of related regions’ function.

ApoB‐related Sulcal depth Patterns Contributed to Executive Function and Plasma Biomarkers of Alzheimer’s disease

AbstractBackgroundAs the cerebral cortex is an important indicator of the pathological changes of AD, ApoB, ApoA1, ApoB/ApoA1 and blood Alzheimer’s biomarkers have been proved to be important markers of AD.MethodWe analyzed data from 21 patients with SCD enrolled in the 500 nm light therapy cohort who filled a battery of neuropsychological tests and had apolipoproteins, plasma Alzheimer’s biomarkers, and gene levels assessed at the time of their baseline 3T MRI scans. The sulcal depth and cortical thickness were obtained using the vertex‐based approach with the CAT12 toolbox (in SPM12).ResultAt corrected thresholds (p < 0.05, FWE‐corrected), there were significant positive correlations between ApoB and sulcal depth in the lateral occipital cortex, and between ApoB/ApoA1 and sulcal depth in the left lateral occipital cortex. In addition, we found a significant negative correlation between shape trails test part B (STT‐B) time and sulcal depth in the left lateral occipital cortex (β = ‐0.502, p = 0.040). Moreover, the plasma Aβ42 was positively related to sulcal depth (β = 0.493, p = 0.044) and plasma p‐tau181 was negatively related to the sulcal depth (β = ‐0.592, p = 0.012) in the left lateral occipital cortex. There was no relationship between cortical thickness and apolipoproteins or any of the biomarkers.ConclusionApoB and ApoB/ApoA1 influence sulcal depth in the left lateral occipital cortex where the sulcal depth may relate to the executive function and plasma Alzheimer’s biomarkers in SCD.

Association of genetic and sulcal traits with executive function in congenital heart disease.

Persons with congenital heart disease (CHD) are at increased risk of neurodevelopmental disabilities, including impairments to executive function. Sulcal pattern features correlate with executive function in adolescents with single-ventricle heart disease and tetralogy of Fallot. However, the interaction of sulcal pattern features with genetic and participant factors in predicting executive dysfunction is unknown. We studied sulcal pattern features, participant factors, and genetic risk for executive function impairment in a cohort with multiple CHD types using stepwise linear regression and machine learning. Genetic factors, including predicted damaging de novo or rare inherited variants in neurodevelopmental disabilitiesrisk genes, apolipoprotein E genotype, and principal components of sulcal pattern features were associated with executive function measures after adjusting for age at testing, sex, mother's education, and biventricular versus single-ventricle CHD in a linear regression model. Using regression trees and bootstrap validation, younger participant age and larger alterations in sulcal pattern features were consistently identified as important predictors of decreased cognitive flexibility with left hemisphere graph topology often selected as the most important predictor. Inclusion of both sulcal pattern and genetic factors improved model fit compared to either alone. We conclude that sulcal measures remain important predictors of cognitive flexibility, and the model predicting executive outcomes is improved by inclusion of potential genetic sources of neurodevelopmental risk. If confirmed, measures of sulcal patterning may serve as early imaging biomarkers to identify those at heightened risk for future neurodevelopmental disabilities.

Association between placental oxygen transport and fetal brain cortical development: a study in monochorionic diamniotic twins.

Normal cortical growth and the resulting folding patterns are crucial for normal brain function. Although cortical development is largely influenced by genetic factors, environmental factors in fetal life can modify the gene expression associated with brain development. As the placenta plays a vital role in shaping the fetal environment, affecting fetal growth through the exchange of oxygen and nutrients, placental oxygen transport might be one of the environmental factors that also affect early human cortical growth. In this study, we aimed to assess the placental oxygen transport during maternal hyperoxia and its impact on fetal brain development using MRI in identical twins to control for genetic and maternal factors. We enrolled 9 pregnant subjects with monochorionic diamniotic twins (30.03 ± 2.39 gestational weeks [mean ± SD]). We observed that the fetuses with slower placental oxygen delivery had reduced volumetric and surface growth of the cerebral cortex. Moreover, when the difference between placenta oxygen delivery increased between the twin pairs, sulcal folding patterns were more divergent. Thus, there is a significant relationship between placental oxygen transport and fetal brain cortical growth and folding in monochorionic twins.

Orbitofrontal sulcal patterns in catatonia.

Catatonia is a psychomotor syndrome frequently observed in disorders with neurodevelopmental impairments, including psychiatric disorders such as schizophrenia. The orbitofrontal cortex (OFC) has been repeatedly associated with catatonia. It presents with an important interindividual morphological variability, with three distinct H-shaped sulcal patterns, types I, II, and III, based on the continuity of the medial and lateral orbital sulci. Types II and III have been identified as neurodevelopmental risk factors for schizophrenia. The sulcal pattern of the OFC has never been investigated in catatonia despite the role of the OFC in the pathophysiology and the neurodevelopmental component of catatonia. In this context, we performed a retrospective analysis of the OFC sulcal pattern in carefully selected homogeneous and matched subgroups of schizophrenia patients with catatonia (N=58) or without catatonia (N=65), and healthy controls (N=82). Logistic regression analyses revealed a group effect on OFC sulcal pattern in the left (χ2=18.1; p<.001) and right (χ2=28.3; p<.001) hemispheres. Catatonia patients were found to have more type III and less type I in both hemispheres compared to healthy controls and more type III on the left hemisphere compared to schizophrenia patients without catatonia. Because the sulcal patterns are indirect markers of early brain development, our findings support a neurodevelopmental origin of catatonia and may shed light on the pathophysiology of this syndrome.

A revised perspective on the evolution of the lateral frontal cortex in primates.

Detailed neuroscientific data from macaque monkeys have been essential in advancing understanding of human frontal cortex function, particularly for regions of frontal cortex without homologs in other model species. However, precise transfer of this knowledge for direct use in human applications requires an understanding of monkey to hominid homologies, particularly whether and how sulci and cytoarchitectonic regions in the frontal cortex of macaques relate to those in hominids. We combine sulcal pattern analysis with resting-state functional magnetic resonance imaging and cytoarchitectonic analysis to show that old-world monkey brains have the same principles of organization as hominid brains, with the notable exception of sulci in the frontopolar cortex. This essential comparative framework provides insights into primate brain evolution and a key tool to drive translation from invasive research in monkeys to human applications.

Development of cortical folds in the human brain: An attempt to review biological hypotheses, early neuroimaging investigations and functional correlates

The folding of the human brain mostly takes place in utero, making it challenging to study. After a few pioneer studies looking into it in post-mortem foetal specimen, modern approaches based on neuroimaging have allowed the community to investigate the folding process in vivo, its normal progression, its early disturbances, and its relationship to later functional outcomes. In this review article, we aimed to first give an overview of the current hypotheses on the mechanisms governing cortical folding. After describing the methodological difficulties raised by its study in fetuses, neonates and infants with magnetic resonance imaging (MRI), we reported our current understanding of sulcal pattern emergence in the developing brain. We then highlighted the functional relevance of early sulcal development, through recent insights about hemispheric asymmetries and early factors influencing this dynamic such as prematurity. Finally, we outlined how longitudinal studies have started to relate early folding markers and the child’s sensorimotor and cognitive outcome. Through this review, we hope to raise awareness on the potential of studying early sulcal patterns both from a fundamental and clinical perspective, as a window into early neurodevelopment and plasticity in relation to growth in utero and postnatal environment of the child.

The Australian gulf snapping turtle Elseya lavarackorum (Testudines: Chelidae) revisited—Is the late Pleistocene fossil species extant?

Disagreement exists on the taxonomic identity of the extant populations of the AustralianElseyareferred to in 1992 as the gulfElseya(= Elseya sp. aff. dentata [Nicholson]). The extant form has since 1997 been considered conspecific with the late Pleistocene fossilElseya lavarackorum(White and Archer, 1994). Recently it has been considered a new species,Elseya oneirosJoseph-Ouni et al., 2020, conspecific with another fossil found in the same site and stratum asElseya lavarackorum. Here we re-examine the fossil material and reassess the characters used by previous authors in an attempt to decide the issue. We find that the anterior bridge suture with the carapace of the fossilElseya lavarackorumis associated with extensive and prominent plastral elements, which has led to misinterpretation of characters associated with this structure. We furthermore show that interindividual variation in sulci patterns is so great as to render them of little taxonomic value. On the basis of (a) deviation of the anterior shape of the carapace from ovoid such that, in aged individuals, the most anterior point of the carapace occurs at marginal scutes M2 (a resultant nuchal bay occurs in such individuals); (b) the typical absence of a cervical scute; (c) no evidence of a medial constriction in the anterior bridge strut suture; and (d) absence of evidence of any other informative variation of taxonomic value; we conclude that the decision to consider the late Pleistocene (ca 23 kyr old) fossil and the extant Elseya sp. aff. dentata [Nicholson] asElseya lavarackorum(White and Archer, 1994) as conspecific should stand.

Exploring the emergence of morphological asymmetries around the brain's Sylvian fissure: a longitudinal study of shape variability in preterm infants.

Brain folding patterns vary within the human species, but some folding properties are common across individuals, including the Sylvian fissure's inter-hemispheric asymmetry. Contrarily to the other brain folds (sulci), the Sylvian fissure develops through the process of opercularization, with the frontal, parietal, and temporal lobes growing over the insular lobe. Its asymmetry may be related to the leftward functional lateralization for language processing, but the time course of these asymmetries' development is still poorly understood. In this study, we investigated refined shape features of the Sylvian fissure and their longitudinal development in 71 infants born extremely preterm (mean gestational age at birth: 26.5weeks) and imaged once before and once at term-equivalent age (TEA). We additionally assessed asymmetrical sulcal patterns at TEA in the perisylvian and inferior frontal regions, neighbor to the Sylvian fissure. While reproducing renowned strong asymmetries in the Sylvian fissure, we captured an early encoding of its main asymmetrical shape features, and we observed global asymmetrical shape features representative of a more pronounced opercularization in the left hemisphere, contrasting with the previously reported right hemisphere advance in sulcation around birth. This added novel insights about the processes governing early-life brain folding mechanisms, potentially linked to the development of language-related capacities.

Comprehensive quantitative analyses of fetal magnetic resonance imaging in isolated cerebral ventriculomegaly

Isolated cerebral ventriculomegaly (IVM) is the most common prenatally diagnosed brain anomaly occurs in 0.2–1 % of pregnancies. However, knowledge of fetal brain development in IVM is limited. There is no prenatal predictor for IVM to estimate individual risk of neurodevelopmental disability occurs in 10 % of children. To characterize brain development in fetuses with IVM and delineate their individual neuroanatomical variances, we performed comprehensive post-acquisition quantitative analysis of fetal magnetic resonance imaging (MRI). In volumetric analysis, brain MRI of fetuses with IVM (n = 20, 27.0 ± 4.6 weeks of gestation, mean ± SD) had revealed significantly increased volume in the whole brain, cortical plate, subcortical parenchyma, and cerebrum compared to the typically developing fetuses (controls, n = 28, 26.3 ± 5.0). In the cerebral sulcal developmental pattern analysis, fetuses with IVM had altered sulcal positional (both hemispheres) development and combined features of sulcal positional, depth, basin area, in both hemispheres compared to the controls. When comparing distribution of similarity index of individual fetuses, IVM group had shifted toward to lower values compared to the control. About 30 % of fetuses with IVM had no overlap with the distribution of control fetuses. This proof-of-concept study shows that quantitative analysis of fetal MRI can detect emerging subtle neuroanatomical abnormalities in fetuses with IVM and their individual variations.

Morphology of Anterior Cingulate Cortex and Its Relation to Schizophrenia.

Cortical folding of the anterior cingulate cortex (ACC), particularly the cingulate (CS) and the paracingulate (PCS) sulci, represents a neurodevelopmental marker. Deviations in in utero development in schizophrenia can be traced using CS and PCS morphometry. In the present study, we measured the length of CS, PCS, and their segments on T1 MRI scans in 93 patients with first- episode schizophrenia and 42 healthy controls. Besides the length, the frequency and the left-right asymmetry of CS/PCS were compared in patients and controls. Distribution of the CS and PCS morphotypes in patients was different from controls. Parcellated sulcal pattern CS3a in the left hemisphere was longer in patients (53.8 ± 25.7 mm vs. 32.7 ± 19.4 mm in controls, p < 0.05), while in CS3c it was reversed—longer in controls (52.5 ± 22.5 mm as opposed to 36.2 ± 12.9 mm, n.s. in patients). Non parcellated PCS in the right hemisphere were longer in patients compared to controls (19.4 ± 10.2 mm vs. 12.1 ± 12.4 mm, p < 0.001). Therefore, concurrent presence of PCS1 and CS1 in the left hemisphere and to some extent in the right hemisphere may be suggestive of a higher probability of schizophrenia.

Cingulate sulcus morphology and paracingulate sulcus variations: Anatomical and radiological studies.

The sulci and gyri found across the cerebrum differ in morphology between individuals. The cingulate sulcus is an important landmark for deciding the surgical approach for neighboring pathological lesions. Identifying the anatomical variations of anterior cingulate cortex morphology would help to determine the safe-entry route through neighboring lesions. In this study, magnetic resonance imaging data acquired from 149 healthy volunteers were investigated retrospectively for anatomical variations of the paracingulate sulcus. Also, human cadaveric brain hemispheres were investigated for cingulate and paracingulate sulcus anatomy. All participants had cingulate sulci in both hemispheres (n=149, 100%). Three types of paracingulate sulcus patterns were identified: "prominent," "present," and "absent." Hemispheric comparisons indicated that the paracingulate sulcus is commonly "prominent" in the left hemisphere (n=48, 32.21%) and more commonly "absent" in the right hemisphere (n=73, 48.99%). Ten (6.71%) people had a prominent paracingulate sulcus in both the right and left hemispheres. Seven (4.70%) of them were male, and 3 (2.01%) of them were female. Paracingulate sulci were present in both hemispheres in 19 people (12.75%), of which 9 (6.04%) were male and 10 (6.71%) were female. There were 35 (23.49%) participants without paracingulate sulci in both hemispheres. Eleven (7.38%) were male and 24 (16.11%) were female. There were 73 (48.99%) participants without right paracingulate sulcus and 57 (38.26%) participants without left paracingulate sulcus (p=0.019). In the examinations of the cadaver hemispheres, the paracingulate sulcus was present and prominent in 25%, and the intralimbic sulcus was present in 15%. It has been observed that the paracingulate sulcus is more prominent in the normal male brain compared to females. In females, there were more participants without paracingulate sulcus. This study shows that there are both hemispheric and sex differences in the anatomy of the paracingulate sulcus. Understanding the cingulate sulcus anatomy and considering the variations in the anterior cingulate cortex morphology during surgery will help surgeons to orient this elegant and complex area.

The brain of Homo habilis: Three decades of paleoneurology

In 1987, Phillip Tobias published a comprehensive anatomical analysis of the endocasts attributed to Homo habilis, discussing issues dealing with brain size, sulcal patterns, and vascular traces. He suggested that the neuroanatomy of this species evidenced a clear change toward many cerebral traits associated with our genus, mostly when concerning the morphology of the frontal and parietal cortex. After more than 30 years, the fossil record associated with this taxon has not grown that much, but we have much more information on cranial and brain biology, and we are using a larger array of digital methods to investigate the paleoneurological variation observed in the human genus. Brain volume, the size of the frontal lobe, or the gross hemispheric asymmetries are still relevant issues, but they are considered to be less central than before. More attention is instead being paid to the cortical organization, the relationships with the cranial architecture, and the influence of molecular or ecological factors. Although the field of paleoneurology can currently count on a larger range of tools and principles, there is still a general lack of anatomical information on many endocranial traits. This aspect is probably crucial for the agenda of paleoneurology. More importantly, the whole science is undergoing a delicate change, because of the growing influence of the social environment. In this sense, the disciplines working with fossils (and, in particular, with brain evolution) should take particular care to maintain a healthy professional situation, avoiding an excess of speculation and overstatement.

The relationship between reading abilities and the left occipitotemporal sulcus: A dual perspective study

Reading activates a region within the left lateral occipitotemporal sulcus (OTS) known as the ‘visual word form area’ (VWFA). While several studies have investigated the impact of reading on brain structure through neuroplastic mechanisms, it has been recently suggested that individual differences in the pattern of the posterior OTS may predict reading skills in adults. In the present study, we first examined whether the structure and morphology and the anatomical connectivity of the left OTS are associated to reading ability. Second, we explored whether reading skills are predicted by the pattern of the left OTS. We found that reading skills were positively associated with increased connectivity between the left OTS and a network of reading-related regions in the left hemisphere. On the other hand, we did not observe an association between the pattern of the left OTS and reading skills. Finally, we found evidence that the morphology and the connectivity of the left OTS are correlated to its sulcal pattern.

Cytoarchitecture, intersubject variability, and 3D mapping of four new areas of the human anterior prefrontal cortex.

The dorsolateral prefrontal cortex (DLPFC) plays a key role in cognitive control and executive functions, including working memory, attention, value encoding, decision making, monitoring, and controlling behavioral strategies. However, the relationships between this variety of functions and the underlying cortical areas, which specifically contribute to these functions, are not yet well-understood. Existing microstructural maps differ in the number, localization, and extent of areas of the DLPFC. Moreover, there is a considerable intersubject variability both in the sulcal pattern and in the microstructure of this region, which impedes comparison with functional neuroimaging studies. The aim of this study was to provide microstructural, cytoarchitectonic maps of the human anterior DLPFC in 3D space. Therefore, we analyzed 10 human post-mortem brains and mapped their borders using a well-established approach based on statistical image analysis. Four new areas (i.e., SFS1, SFS2, MFG1, and MFG2) were identified in serial, cell-body stained brain sections that occupy the anterior superior frontal sulcus and middle frontal gyrus, i.e., a region corresponding to parts of Brodmann areas 9 and 46. Differences between areas in cytoarchitecture were captured using gray level index profiles, reflecting changes in the volume fraction of cell bodies from the surface of the brain to the cortex-white matter border. A hierarchical cluster analysis of these profiles indicated that areas of the anterior DLPFC displayed higher cytoarchitectonic similarity between each other than to areas of the neighboring frontal pole (areas Fp1 and Fp2), Broca's region (areas 44 and 45) of the ventral prefrontal cortex, and posterior DLPFC areas (8d1, 8d2, 8v1, and 8v2). Area-specific, cytoarchitectonic differences were found between the brains of males and females. The individual areas were 3D-reconstructed, and probability maps were created in the MNI Colin27 and ICBM152casym reference spaces to take the variability of areas in stereotaxic space into account. The new maps contribute to Julich-Brain and are publicly available as a resource for studying neuroimaging data, helping to clarify the functional and organizational principles of the human prefrontal cortex.