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Interhemispheric connections in neonatal owl monkeys (Aotus trivirgatus) and galagos (Galago crassicaudatus).

Interhemispheric connections were studied by injecting a mixture of horseradish peroxidase (HRP) and wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP) into multiple sites in dorsolateral occipital and parietal cortex of one cerebral hemisphere of three galagos (Galago crassicaudatus) and two owl monkeys (Aotus trivirgatus) within seven days of birth. Cortex was either separated from the rest of the brain, flattened and cut parallel to the surface to aid reconstructing surface-view patterns of labeled neurons and processes, or cut in standard coronal or parasagittal planes to better reveal laminar patterns of connections. In both primate species, the surface-view pattern of callosal connections in infants was remarkably adult-like. In infant owl monkeys, callosal connections were concentrated along the margin of area 18 with area 17, and only a few labeled cells were found within area 17. Other visual areas including the second visual area, V-II, and the middle temporal visual area, MT, had patchy distributions of labeled neurons that extended over large parts of the visual field representations. Primary motor, auditory, and somatosensory fields also had patchy distributions of labeled neurons, with regions of areas 3b and adjoining somatosensory fields having few callosal connections in portions that appeared to correspond with representations of the hand and foot. Results were very similar in galagos, except that newborn galagos, as in adults, had a patchy distribution of callosally projecting neurons that extended well within area 17. Furthermore, the labeled neurons were concentrated in patches that aligned with the cytochrome oxidase blobs of area 17. Finally, callosal connections were concentrated in cytochrome oxidase poor regions of area 3b.

Animals↗

Reproduction in the Mexican leaf frog, Pachymedusa dacnicolor. II. The male.

The Mexican leaf frog, Pachymedusa dacnicolor, an inhabitant of the semiarid, subtropical Mexican lowlands, displays a well-defined seasonal testicular cycle. Testis weight seems to be a reliable index of the reproductive status of the animal and plasma levels of androgens (testosterone, T; 5 alpha-dihydrotestosterone, DHT; androstenedione, A) correlate not only with testicular growth, but with callosity development, reproductive behavior, and breeding. During the fall and winter, testis weight reaches its minimum as do plasma concentrations of T, DHT, A, and estradiol-17 beta (E). Plasma levels of progesterone (P) are maintained at a very low level throughout the year. During the fall and winter, spermatogenesis is almost entirely absent and the callosities are white and smooth. No signs of reproductive behavior (calling and amplexus) are evident. Late spring marks the initiation of spermatogenesis, testis weight increase, darkening of callosities, and increase in plasma levels of androgens. The magnitude of callosity development and the onset of calling and amplectant behavior are correlated with a great rise in plasma androgen levels. Although plasma T concentrations were higher than plasma levels of DHT and A, this androgen failed to stimulate the development of callosities and calling behavior in successfully castrated males.

Androgens↗

Developmental changes in the heavy subunit of neurofilaments in the corpus callosum of the cat.

In the corpus callosum of the cat, the heavy subunit of neurofilaments (NFH) can be demonstrated with the monoclonal antibody NE14, as early as P11, not at P3, and only in a few axons. At P18-19 and more markedly at P29, many more callosal axons have become positive to NE14 and this is similar to what is found in the adult. In contrast, callosal axons become positive to the neurofilament antibody SMI-32 only between P29 and P39 and remain positive in the adult. Treatment with alkaline phosphatase prevents axonal staining with NE14, but results in SMI-32 staining of a few callosal axons as early as P11, but not at P3. Between P11 and P19 the number of axons stained with SMI-32 after alkaline phosphatase treatment increases, in parallel with that of axons stained with NE14. Thus NE14 appears to recognize a phosphorylated form of NFH, while SMI-32 appears to recognize an epitope of NFH which is either masked by phosphate or inaccessible until between P29 and P39, unless the tissue is treated with alkaline phosphatase. These two forms of NFH appear towards the end of the period of massive developmental elimination of callosal axons. They are also synchronous with changes in the spacing of neurofilaments quantified in a separate ultrastructural study. These cytoskeletal changes may terminate the juvenile-labile state of callosal axons and allow further axial growth of the axon.

Aging↗

Developmental interactions between the corpus callosum and the visual system in cats.

The neonatal corpus callosum is involved in the development of pathways or mechanisms that coordinate the inputs from the two eyes. Several related visual functions are permanently altered by the absence of the callosum during early development. As determined behaviorally and by visual evoked potentials, the normal amount of 90 degrees of the visual field in which both eyes respond to stimulation is almost completely eliminated. Also, there is a reduced number of binocular cells in striate cortical regions representing most of the visual field. In addition, behaviorally measured visual acuity is reduced. Changes in acuity and striate binocularity only result when a callosal section occurs during a critical period of the first postnatal month, and the earlier the surgery, the greater the changes. The lack of myelination during the first postnatal month indicates that the conduction properties of the callosum are poorly developed during its critical period. The pattern of callosal connectivity is probably significant for its role in development, but not all callosal fibers are necessary for normal visual development. Developmental plasticity of callosal connections has been demonstrated for striate cortex, but now it has also been demonstrated for the claustrum. Thus, the callosal role in regions representing both central and peripheral visual field, in neocortical and non-neocortical brain areas should be reassessed.

Aging↗

Cellular organisation in meiotic and early post-meiotic rice anthers.

We have used fluorescent, confocal laser and transmission electron microscopy (TEM) to examine cellular organisations, including callose (1,3-beta-glucan) behaviour, in meiotic and early post-meiotic rice anthers. These features are critical for pollen formation and provide information to better understand pollen sterility caused by abiotic stress in rice and other monocotyledonous species. Among organelles during meiosis, abundant plastids, mitochondria and nuclei of the anther cells show distinctive features. Chloroplasts in the endothecium store starch and indicate a potential for photosynthetic activity. During meiosis, the middle layer cells are markedly compressed and at the tetrad stage are either vacuolated or filled with degenerating electron-opaque organelles. Viable mitochondria, stained with Rhodamine 123, are seen in the endothecium and tapetum, but the mitochondria in the middle layer are not stained during meiosis. The radial walls of the tapetum are disorganised and degenerating, indicating the formation of a syncytium; pro-orbicules are located at the locular walls at the tetrad stage. Immunohistochemical studies show that the sporogenous cells are entirely enveloped by a thick callosic layer at early meiosis. Cell plate callose was assembled in a plane between the dyad cells. In the tetrads, however, callose formed only at the centre, showing that the tetrad microspores are not enveloped but separated by callose walls. Thick, undulating electron-opaque walls around the tetrads indicate the beginning of exinous microspore wall differentiation.

Aniline Compounds↗

Relationship between attentional performance and corpus callosum morphometry in patients with Alzheimer's disease.

There has been considerable interest in cognitive deficits associated with Alzheimer's disease (AD) and relationships between these impairments and specific cortical atrophies. Two previous studies [Neuropsychologia 28 (1990) 1197; Dementia 3 (1992) 350] have found that AD patients exhibit significant impairments in the attentional ID/ED set-shifting tasks of the CANTAB battery which involved attentional shifting abilities. But, at present, no study has examined the neural bases of these abilities in AD patients. In the present study, the relationship between performances on this attentional test and morphometry of the anterior and posterior corpus callosum is examined in AD patients in the mild to moderate stages of the disease (n=30, mean age=74.1+/-4.9 years, mean MMSE score=23.9+/-2.6). A control group is constituted (n=20, mean age=73.15+/-5.5 years) for comparison of cerebral measurements. The stepwise multiple regression analysis indicates that the relative contribution for the total callosal and the anterior CC areas of the simple discrimination subtest is significantly positive whereas for the posterior callosal areas the relative contribution of the more complex subtest is significantly positive. AD patients from the subgroup "low", who failed to do the nine subtests of the attentional set-shifting tasks, exhibit smaller callosal areas than control subjects. There is no significant difference for all callosal measurements between AD patients from the subgroup "high", who completely succeeded the test, and control subjects. Our findings suggest that the anterior corpus callosum would be related to attentional shifting abilities in AD patients. Moreover, these patients with probable AD seem heterogeneous for performances in the attentional test of the CANTAB and for callosal atrophies.

Aged↗

The size of corpus callosum and functional connectivities of cortical regions in finger and shoulder movements.

The correlations between the size of corpus callosum and the inter- and intra-hemispheric EEG coherence and the spatial EEG synchronization during finger and shoulder movements were analyzed in nine right-handed men. The cross-sectional surface areas of corpus callosum (CC) and of seven callosal regions were measured from the mid-sagittal slice of the anatomical MRI. Movement-related coherence between pairs of EEG electrodes overlying the central and parietal regions of both hemispheres was computed after spatially filtering EEG data by the Laplacian operator method. The spatial EEG synchronization was evaluated using omega-complexity, a novel measure which quantifies the number of independent sources of spontaneous EEG oscillations. The amplitude of coherence between the left and right S1/M1 areas after movement onset in the lower alpha band (7.8-9.8 Hz) correlated with the size of the callosal body in both types of movement. The size of the callosal body also correlated with the C3-Cz coherence in the 15.6-19.5 Hz band in finger movement, and in the 15.6-23.5 Hz band in shoulder movements. The size of the rostral, anterior intermediate and posterior intermediate truncus of CC correlated with omega-complexity in both types of movements indicating more foci of synchronized EEG oscillations in subjects with a large callosal truncus. The results suggest that the size of callosal truncus which is known to connect the primary sensorimotor and the supplementary motor areas of both hemispheres contributes to the coupling of EEG oscillations during voluntary finger and shoulder movements.

Adult↗

Isolation of aluminum-tolerant cell lines of tobacco in a simple calcium medium and their responses to aluminum.

Aluminum (Al)-tolerant cell lines (ALT301 and ALT401) of tobacco were isolated in a simple calcium (Ca) solution from ethyl methane sulfonate (EMS)-treated suspension cultured tobacco cells (Nicotiana tabacum L. cv. Samsun, a cell line SL) at the logarithmic phase of growth. A highly tolerant cell line ALT301 exhibited the accumulation of Al and the deposition of callose to the same extent as the parental SL cells during the exposure to Al. However, the Al-treated ALT301 cells grew much better than the Al-treated SL cells after transfer to Al-free growth medium. Compared to SL cells, ALT301 cells were more tolerant to toxicity of copper and iron, but not to that of lanthanum. These results suggest that ALT301 cells have an internal tolerance mechanism, which makes cells grow normally in spite of Al accumulation and Al-induced lesion represented by the deposition of callose. This tolerance mechanism seems also to be effective against copper and iron toxicity. A slightly tolerant cell line ALT401 also accumulated Al to the same degree as SL cells, but deposited significantly less callose than did SL cells (43% of SL). The growth of ALT401 cells after Al treatment was only slightly better than that of SL cells. Thus, it seems that ALT401 cells have a mechanism to protect cells only from the Al-induced deposition of callose, which is not enough to overcome the Al-induced inhibition of growth. The different phenotypes exhibited by these Al-tolerant cell lines suggest that the deposition of callose is not directly related to the inhibition of growth in Al-treated cells.

Journal Article↗

Arabidopsis mutants with increased sensitivity to aluminum.

Al-sensitive (als) mutants of Arabidopsis were isolated and characterized with the aim of defining mechanisms of Al toxicity and resistance. Most als mutants selected on the basis of root growth sensitivity to Al were recessive, and together the mutants constituted eight complementation groups. Also, in most als mutants, Al sensitivity appeared to be specific for Al relative to La (another trivalent cation), except als2, which was more sensitive to La than wild type. The tendency of roots on mutant seedlings to accumulate Al was examined by staining with morin and hematoxylin, dyes used to indicate the presence of Al. A significant increase in morin staining was observed in als5, consistent with its increased sensitivity to Al. Unexpectedly, als7 and als4 showed less morin staining, suggesting that the roots on these mutants accumulate less Al than wild type seedlings after exposure to Al-containing solutions. Roots of wild-type seedlings produce callose in response to AlCl3 concentrations that inhibit root growth. Only als5 accumulated more callose than wild type in response to low levels (25 mu M) of AICI3 However, als4 and als7 did not accumulate callose at this AlCl3 concentration even though root growth was significantly inhibited. The lack of callose accumulation in als4 and als7 suggests that there is not an obligatory relationship between callose deposition and Al-induced inhibition of root growth.

Aluminum↗

beta-Furfuryl-beta-Glucoside: An Endogenous Activator of Higher Plant UDP-Glucose:(1-3)-beta-Glucan Synthase : Biological Activity, Distribution, and in Vitro Synthesis.

In a recent paper (P Ohana, DP Delmer, JC Steffens, DE Matthews, R Mayer, M Benziman [1991] J Biol Chem 266: 13472-13475), we described the purification and structural characterization of beta-furfuryl-beta-glucoside (FG), an endogenous activator of plant UDP-glucose:(1-->3)-beta-glucan (callose) synthase. In the present report, we provide evidence that FG specifically stimulates callose synthase. The effects of FG on the kinetic properties of callose synthase were studied, and we ascertained that FG, or at least a very similar compound, is present in other plant systems. Chemically synthesized alpha-furfuryl-beta-glucoside also stimulates callose synthase, exhibiting a slightly higher K(a) of 80 micromolar, compared with 50 micromolar for FG. In addition, we have identified and partially characterized an enzyme that catalyzes the synthesis of FG using beta-furfuryl alcohol and UDP-glucose as substrates. A model for the regulation of callose synthesis in vivo, involving changes in intracellular compartmentation of FG and Ca(2+), is proposed.

Journal Article↗

Responses of cortical neurons to stimulation of corpus callosum in vitro.

1. An in vitro slice preparation of rat cingulate cortex was used to analyze the responses of layer V neurons to electrical stimulation of the corpus callosum (CC). In addition, synaptic termination of callosal afferents with layer V neurons was evaluated electron microscopically to provide a structural basis for interpreting some of the observed response sequences. 2. Layer V neurons had a resting membrane potential (RMP) of 60 +/- 0.68 (SE) mV, an input resistance of 47 +/- 4.74 M omega, a membrane time constant of 4.37 +/- 0.51 ms, an electrotonic length constant of 1.38 +/- 0.25, and produced spontaneous action potentials that were 50 +/- 0.3 mV in amplitude. Intracellular depolarizing current pulses evoked spikes that were sometimes associated with low-amplitude (2-5 mV) depolarizing (5-10 ms in duration) and hyperpolarizing (10-20 ms in duration) afterpotentials. 3. A single stimulus of increasing intensities to the CC produced one of the following response sequences: a) antidromic spike and an excitatory postsynaptic potential (EPSP), which initiated one or more spikes; b) antidromic spike, EPSP-evoked action potentials, and a hyperpolarization, which may have represented an intrinsic cell property or inhibitory synaptic activity; c) EPSP and evoked spikes only; d) high-amplitude EPSP with an all-or-none burst of action potentials. 4. Antidromically activated (AA) neurons always produced EPSPs in response to CC stimulation. When compared with nonantidromically activated neurons, AA cells had a more negative RMP, greater electrotonic length constant (LN), higher ratio of dendritic to somatic conductance (rho), and formed shorter duration, callosal-evoked EPSPs. 5. Neurons in anterior cingulate cortex produced EPSPs of longer duration than did those in posterior cortex (50 +/- 3.57 versus 26 +/- 1.56 ms, respectively). EPSPs in anterior neurons also had a higher maximum amplitude (20.5 +/- 1.0 versus 11.5 +/- 0.79 mV) and longer time to peak (11.6 +/- 2.2 versus 8.2 +/- 0.8 ms). 6. Electron microscopy of Golgi-impregnated neurons following contralateral lesions demonstrated that both pyramidal and nonpyramidal neurons received direct callosal afferents. Synaptic termination of callosal axons with the apical dendritic trees of anterior pyramidal cells was 6 times greater than it was with posterior pyramidal neurons. 7. EPSP shape differences in anterior and posterior neurons may be partially accounted for by the density and distribution of callosal afferents to these two cortices.

Animals↗

Species differences and similarities in the fine structure of the mammalian corpus callosum.

A cross-species ultrastructural study of the corpus callosum was performed in six domestic species: the rat, the rabbit, the cat, the dog, the horse and the cow. The results indicate cross-species conservatism in callosal fiber composition with a good interspecies relation between fiber number and brain size. Across species, increases in both brain size and callosal area indicate more callosal fibers, although less than expected from the estimated increase in cortical cell number. Within each species, the correlation between fiber number and brain weight tends to disappear, although in most cases a larger callosum implies a larger number of callosal fibers. The median fiber diameter was conservative across species (0.11-0.2 microm), indicating the maintenance of conduction velocity of most callosal fibers regardless of interhemispheric distance. Nevertheless, the maximal fiber diameters tended to be higher in species with larger brains. Therefore, there is a population of coarse-diameter fibers that tend to increase their diameter and conduction velocity with increasing brain size. However, allometric calculations suggest that the associated increase in velocity in these large fibers may not be sufficient to maintain a constant interhemispheric transmission time in different species.

Animals↗

Magnetic resonance imaging of the corpus callosum: predictors of size in normal adults.

Eighty normal adults were studied with MRI to investigate the relationship between regional morphology of the corpus callosum and characteristics such as age, gender, education, and cranial size. The variability coefficient was 20% in total callosal area and from 19% to 40% in regional callosal area. Increasing age and smaller cranial area were both associated with smaller total and regional callosal areas; there were no effects of gender and education. The relative effects of age and cranial size varied across regions and were most prominent for anterior subdivisions. However, age and cranial size together explained less than half the variance in regional callosal size. Further study is needed to identify additional correlates of regional callosal anatomy.

Adult↗

Sonographic biometry of the frontal lobe in normal and growth-restricted neonates.

Assessing the impact of restricted intrauterine growth on neonatal frontal lobe (FL) dimensions is important. We aimed to create a sonographic nomogram of FL dimensions in neonates at different gestational ages (GA) and evaluate the impact of small head circumference (HC) on FL dimensions. We conducted sonographic biometry of the FL at birth. We included 218 newborn infants born at GA of 24-43 wk: appropriate for GA and normal HC (n = 178), and small for GA and small HC (n = 23). Infants with a 5-min Apgar score <7, severe congenital malformations, or chromosomal abnormalities were excluded. Through a coronal ultrasound scan via the anterior fontanelle at the level where the most lateral point of the left Sylvian fissure was best demonstrated, we drew a triangle connecting the most lateral point of the Sylvian fissure, the corpus callosum, and the subcalvarian point of the interhemispheric fissure. We measured the three sides of the triangle, Sylvian-fontanellar distance, Sylvian-callosal distance, and fontanellar-callosal distance, and calculated the frontal triangular area. All four FL dimensions increased significantly between 24 and 43 wk of gestation in both appropriate for GA-normal HC and small for GA-small HC neonates, and were strongly correlated with HC and birth weight. Regression lines of GA against Sylvian-fontanellar distance, Sylvian-callosal distance, fontanellar-callosal distance, and frontal triangular area in the appropriate for GA-normal HC group differed significantly from those of the small for GA-small HC group (p < 0.05). Male neonates had significantly larger Sylvian-fontanellar and Sylvian-callosal distances than females (p < 0.01 and p < 0.015, respectively). In conclusion, FL measures increased significantly between 24 and 43 wk of gestation, and were strongly correlated with HC. We speculate that a sonographically small fetal HC implies growth restriction of the fetal FL.

Age Factors↗

The Wnt receptor Ryk is required for Wnt5a-mediated axon guidance on the contralateral side of the corpus callosum.

Ryk (receptor related to tyrosine kinase) has been shown to be a novel Wnt receptor in both Caenorhabditis elegans and Drosophila melanogaster. Recently, Ryk-Wnt interactions were shown to guide corticospinal axons down the embryonic mouse spinal cord. Here we show that, in Ryk-deficient mice, cortical axons project aberrantly across the major forebrain commissure, the corpus callosum. Many mouse mutants have been described in which loss-of-function mutations result in the inability of callosal axons to cross the midline, thereby forming Probst bundles on the ipsilateral side. In contrast, loss of Ryk does not interfere with the ability of callosal axons to cross the midline but impedes their escape from the midline into the contralateral side. Therefore, Ryk(-/-) mice display a novel callosal guidance phenotype. We also show that Wnt5a acts as a chemorepulsive ligand for Ryk, driving callosal axons toward the contralateral hemisphere after crossing the midline. In addition, whereas callosal axons do cross the midline in Ryk(-/-) embryos, they are defasciculated on the ipsilateral side, indicating that Ryk also promotes fasciculation of axons before midline crossing. In summary, this study expands the emerging role for Wnts in axon guidance and identifies Ryk as a key guidance receptor in the establishment of the corpus callosum. Our analysis of Ryk function further advances our understanding of the molecular mechanisms underlying the formation of this important commissure.

Animals↗

Fetal magnetic resonance imaging in the evaluation of fetuses referred for sonographically suspected abnormalities of the corpus callosum.

OBJECTIVE: Fetal magnetic resonance imaging (MRI) has been shown to be useful in assessing the developing central nervous system. However, its utility in specific brain disorders has not been well investigated. We hypothesized that fetal MRI can better assess the integrity of the brain in cases with sonographically suspected callosal abnormalities. METHODS: We retrospectively reviewed fetal MRI and prenatal sonographic studies of 10 fetuses referred for MRI for sonographically suspected callosal abnormalities. RESULTS: An abnormal corpus callosum was identified on fetal MRI in 80% of cases. The type of callosal abnormality (complete or partial agenesis) was similar on both prenatal sonography and fetal MRI in all cases. All sonographically identified additional brain abnormalities were detected on fetal MRI, with the exception of choroid plexus cysts. Furthermore, in 63% (5 of 8) of cases with a callosal abnormality on both sonography and fetal MRI, additional brain abnormalities were detected on fetal MRI that were not apparent on sonography. These sonographically occult findings were confirmed on postnatal MRI or autopsy in 3 of 5 patients. CONCLUSIONS: Fetal MRI is an important adjunct to sonography in assessing the corpus callosum and other aspects of brain development when agenesis of the corpus callosum is suspected. It can identify frequent additional findings that are not visible on sonography such as abnormal sulcation. In light of the association between additional brain abnormalities and worse neurodevelopmental outcome, the potential of fetal MRI as an important adjunctive prognostic imaging test in fetuses with callosal agenesis can now be tested.

Agenesis of Corpus Callosum↗

[Agenesis of the corpus callosum. Neuropathologic study and physiopathologic hypotheses].

The neuropathological study of corpus callosum agenesis requires a two-phase approach: first it should analyze the putative causal factors, i.e. absence of callosal neurons, commissuration inability or synapse remodelling defect; secondly it has to detect any morphogenetic effects stemming from the absence of commissure such as nonregression of archicortical structures, ventricular enlargement or possible invasion of the remaining telencephaplic commissure by callosal neurons. Absence of callosal neurons due to abnormal corticogenesis gives rise to corpus callosum agenesis without callosal axon, that is without Probst's bundles. Conversely, corpus callosum agenesis occurring secondary to a commissuration default is associated with the presence of callosal axons which travel along the midline instead of crossing, that leads to the formation of Probst's bundles. This inability to cross the midline could be secondary to an obstacle, such as lipoma or as interhemispheric cysts, or primitive due to axonal guidance disturbance. In the latter situation, the commissural defect could affect the other cerebral commissures i.e. anterior or hippocampal commissures, or could become integrated into a more diffuse midline pathology involving both cerebral and extracerebral structures. Finally, it could be assumed that a synapse remodelling defect could lead to atrophy or hypertrophy of the commissure, that occurs in the absence of white matter pathology.

Agenesis of Corpus Callosum↗

Interhemispheric connections of visual cortex in the owl monkey, Aotus trivirgatus, and the bushbaby, Galago senegalensis.

Anatomical techniques have been used to map within visual cortex th pattern of degenerating axonal terminals produced by surgical section of the splenium of the corpus callosum in the owl monkey, Aotus trivirgatus, and the bushbaby, Galago senegalensis. Previous studies in other species have shown that callosal inputs terminate preferentially in regions where the vertical meridian of the visual field is represented. Such a correspondence can serve as a useful aid for locating the boundaries of visual areas. The goals of this study have been (1) to assess the degree of correspondence between callosal inputs and previously identified vertical meridian representations in the owl monkey and bushbaby, and (2) to gain information from the pattern of callosal inputs concerning the existence and organization of as yet unidentified extrastriate visual areas. In both the owl monkey and the bushbaby, a discrete band of degenerating axonal terminals corresponds precisely to the vertical meridian representation at the V1-V2 border, and a less precise increase in the density of degenerating axonal terminals corresponds to the vertical meridian representation of extrastriate area MT. A well-defined band of degeneration on the ventral surface of the owl monkey's cerebral hemisphere corresponds to a previously unknown vertical meridian representation which is shared by two newly identified extrastriate visual areas. Elsewhere in visual cortex the pattern of callosal connections is more complex. Although this pattern may still reflect visual topography, it is not immediately useful for distinguishing areal boundaries.

Animals↗