Cortical connections of the functional domain for climbing or running in posterior parietal cortex of galagos.

Previous studies in prosimian galagos (Otolemur garnetti) have demonstrated that posterior parietal cortex (PPC) is subdivided into several functionally distinct domains, each of which mediates a specific type of complex movements (e.g., reaching, grasping, hand-to-mouth) and has a different pattern of cortical connections. Here we identified a medially located domain in PPC where combined forelimb and hindlimb movements, as if climbing or running, were evoked by long-train intracortical microstimulation. We injected anatomical tracers in this climbing/running domain of PPC to reveal its cortical connections. Our results showed the PPC climbing domain had dense intrinsic connections within rostral PPC and reciprocal connections with forelimb and hindlimb region in primary motor cortex (M1) of the ipsilateral hemisphere. Fewer connections were with dorsal premotor cortex (PMd), supplementary motor (SMA), and cingulate motor (CMA) areas, as well as somatosensory cortex including areas 3a, 3b, and 1-2, secondary somatosensory (S2), parietal ventral (PV), and retroinsular (Ri) areas. The rostral portion of the climbing domain had more connections with primary somatosensory cortex than the caudal portion. Cortical projections were found in functionally matched domains in M1 and premotor cortex (PMC). Similar patterns of connections with fewer labeled neurons and terminals were seen in the contralateral hemisphere. These connection patterns are consistent with the proposed role of the climbing/running domain as part of a parietal-frontal network for combined use of the limbs in locomotion as in climbing and running. The cortical connections identify this action-specific domain in PPC as a more somatosensory driven domain.

A digital collection of rare and endangered lemurs and other primates from the Duke Lemur Center.

Scientific study of lemurs, a group of primates found only on Madagascar, is crucial for understanding primate evolution. Unfortunately, lemurs are among the most endangered animals in the world, so there is a strong impetus to maximize as much scientific data as possible from available physical specimens. MicroCT scanning efforts at Duke University have resulted in scans of more than 100 strepsirrhine cadavers representing 18 species from the Duke Lemur Center. An error study of the microCT scanner recovered less than 0.3% error at multiple resolution levels. Scans include specimen overviews and focused, high-resolution selections of complex anatomical regions (e.g., cranium, hands, feet). Scans have been uploaded to MorphoSource, an online digital repository for 3D data. As captive (but free ranging) individuals, these specimens have a wealth of associated information that is largely unavailable for wild populations, including detailed life history data. This digital collection maximizes the information obtained from rare and endangered animals with minimal degradation of the original specimens.

The Distribution of Ki-67 and Doublecortin-Immunopositive Cells in the Brains of Three Strepsirrhine Primates: Galago demidoff, Perodicticus potto, and Lemur catta.

This study investigated the pattern of adult neurogenesis throughout the brains of three prosimian primate species using immunohistochemical techniques for endogenous markers of this neural process. Two species, Galago demidoff and Perodicticus potto, were obtained from wild populations in the primary rainforest of central Africa, while one species, Lemur catta, was captive-bred. Two brains from each species, perfusion-fixed with 4% paraformaldehyde, were sectioned (50 µm section thickness) in sagittal and coronal planes. Using Ki-67 and doublecortin (DCX) antibodies, proliferating cells and immature neurons were identified in the two canonical neurogenic sites of mammals, the subventricular zone of the lateral ventricle (SVZ) giving rise to the rostral migratory stream (RMS), and the subgranular zone of the dentate gyrus of the hippocampus. In addition a temporal migratory stream (TMS), emerging from the temporal horn of the lateral ventricle to supply the piriform cortex and adjacent brain regions with new neurons, was also evident in the three prosimian species. While no Ki-67-immunoreactive cells were observed in the cerebellum, DCX-immunopositive cells were observed in the cerebellar cortex of all three species. These findings are discussed in a phylogenetic context.

Frontal eye field in prosimian galagos: Intracortical microstimulation and tracing studies.

The frontal eye field (FEF) in prosimian primates was identified as a small cortical region, above and anterior to the anterior frontal sulcus, from which saccadic eye movements were evoked with electrical stimulation. Tracer injections revealed FEF connections with cortical and subcortical structures participating in higher order visual processing. Ipsilateral cortical connections were the densest with adjoining parts of the dorsal premotor and prefrontal cortex (PFC). Label in a region corresponding to supplementary eye field (SEF) of other primates, suggests the existence of SEF in galagos. Other connections were with ventral premotor cortex (PMV), the caudal half of posterior parietal cortex, cingulate cortex, visual areas within the superior temporal sulcus, and inferotemporal cortex. Callosal connections were mostly with the region of the FEF of another hemisphere, SEF, PFC, and PMV. Most subcortical connections were ipsilateral, but some were bilateral. Dense bilateral connections were to caudate nuclei. Densest reciprocal ipsilateral connections were with the paralamellar portion of mediodorsal nucleus, intralaminar nuclei and magnocellular portion of ventral anterior nucleus. Other FEF connections were with the claustrum, reticular nucleus, zona incerta, lateral posterior and medial pulvinar nuclei, nucleus limitans, pretectal area, nucleus of Darkschewitsch, mesencephalic and pontine reticular formation and pontine nuclei. Surprisingly, the superior colliculus (SC) contained only sparse anterograde label. Although most FEF connections in galagos are similar to those in monkeys, the FEF-SC connections appear to be much less. This suggests that a major contribution of the FEF to visuomotor functions of SC emerged with the evolution of anthropoid primates.

A giant among dwarfs: a new species of galago (Primates: Galagidae) from Angola.

OBJECTIVES: Based on vocalization recordings of an unknown galago species, our main objectives were to compare morphology and call structure with known closely-related taxa and describe a new species of galago. MATERIALS AND METHODS: We conducted field surveys in three forest habitats along the escarpment region in western Angola (Kumbira Forest, Bimbe Area, and Northern Scarp Forest), and examined galago specimens from museums worldwide. We digitized and analyzed calls using Avisoft SASLab Pro software. We also compared museum specimens from Angola with other Galago and Galagoides specimens, and conducted comparative analyses (ANOVA and between group principle component analysis) based on a set of twelve linear measurements of skulls and teeth. RESULTS: We describe the new species to which we give the name Angolan dwarf galago, Galagoides kumbirensis sp. nov. The new species has a loud and characteristic crescendo call, used by other Galagoides spp. (sensu stricto) in West Africa to attract companions and repel rivals. However, this call shows species-typical differences from its closest relatives. Galagoides kumbirensis sp. nov. is also distinguished by differences in the skull morphology, pelage color and facial markings, as well as a larger body size, similar to that of Galago moholi, which is not known to be sympatric. CONCLUSION: This discovery points to the importance of Angolan forests as refuges for endemic biodiversity. These forests are under severe threat from overexploitation, and there is an urgent need to establish conservation measures and designate protected areas.

Cortical Connections of the Caudal Portion of Posterior Parietal Cortex in Prosimian Galagos.

Posterior parietal cortex (PPC) of prosimian galagos includes a rostral portion (PPCr) where electrical stimulation evokes different classes of complex movements from different subregions, and a caudal portion (PPCc) where such stimulation fails to evoke movements in anesthetized preparations ( Stepniewska, Fang et al. 2009). We placed tracer injections into PPCc to reveal patterns of its cortical connections. There were widespread connections within PPCc as well as connections with PPCr and extrastriate visual areas, including V2 and V3. Weaker connections were with dorsal premotor cortex, and the frontal eye field. The connections of different parts of PPCc with visual areas were roughly retinotopic such that injections to dorsal PPCc labeled more neurons in the dorsal portions of visual areas, representing lower visual quadrant, and injections to ventral PPCc labeled more neurons in ventral portions of these visual areas, representing the upper visual quadrant. We conclude that much of the PPCc contains a crude representation of the contralateral visual hemifield, with inputs largely, but not exclusively, from higher-order visual areas that are considered part of the dorsal visuomotor processing stream. As in galagos, the caudal half of PPC was likely visual in early primates, with the rostral PPC half mediating sensorimotor functions.

Organization of cholinergic, catecholaminergic, serotonergic and orexinergic nuclei in three strepsirrhine primates: Galago demidoff, Perodicticus potto and Lemur catta.

The nuclear organization of the cholinergic, catecholaminergic, serotonergic and orexinergic systems in the brains of three species of strepsirrhine primates is presented. We aimed to investigate the nuclear complement of these neural systems in comparison to those of simian primates, megachiropterans and other mammalian species. The brains were coronally sectioned and immunohistochemically stained with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. The nuclei identified were identical among the strepsirrhine species investigated and identical to previous reports in simian primates. Moreover, a general similarity to other mammals was found, but specific differences in the nuclear complement highlighted potential phylogenetic interrelationships. The central feature of interest was the structure of the locus coeruleus complex in the primates, where a central compactly packed core (A6c) of tyrosine hydroxylase immunopositive neurons was surrounded by a shell of less densely packed (A6d) tyrosine hydroxylase immunopositive neurons. This combination of compact and diffuse divisions of the locus coeruleus complex is only found in primates and megachiropterans of all the mammalian species studied to date. This neural character, along with variances in a range of other neural characters, supports the phylogenetic grouping of primates with megachiropterans as a sister group.

The hustle and bustle of city life: monitoring the effects of urbanisation in the African lesser bushbaby.

Urbanisation has become a severe threat to pristine natural areas, causing habitat loss and affecting indigenous animals. Species occurring within an urban fragmented landscape must cope with changes in vegetation type as well as high degrees of anthropogenic disturbance, both of which are possible key mechanisms contributing to behavioural changes and perceived stressors. We attempted to elucidate the effects of urbanisation on the African lesser bushbaby, Galago moholi, by (1) recording activity budgets and body condition (body mass index, BMI) of individuals of urban and rural populations and (2) further determining adrenocortical activity in both populations as a measure of stress via faecal glucocorticoid metabolite (fGCM) levels, following successful validation of an appropriate enzyme immunoassay test system (adrenocorticotropic hormone (ACTH) challenge test). We found that both sexes of the urban population had significantly higher BMIs than their rural counterparts, while urban females had significantly higher fGCM concentrations than rural females. While individuals in the urban population fed mainly on provisioned anthropogenic food sources and spent comparatively more time resting and engaging in aggressive interactions, rural individuals fed almost exclusively on tree exudates and spent more time moving between food sources. Although interactions with humans are likely to be lower in nocturnal than in diurnal species, our findings show that the impact of urbanisation on nocturnal species is still considerable, affecting a range of ecological and physiological aspects.

Uterine gland microstructure in the pregnant and the non-pregnant lesser galago (galago senegalensis) / Microestructura de la glándula uterina en la gálago menor (galago senegalensis) preñada y no preñada

The histology and the ultrastructure of the uterine glands of the lesser bushbaby (Galago senegalensis) were studied in six specimens (5 pregnant and one non pregnant) which were fixed partly with bouin's fixative and part with 2.5% glutaraldehyde in 0.1 M cacodylate buffer. An overview of the main results revealed uterine glands in the non-pregnant uterus are rudimentary and scarce within the mucosa. In early pregnancy (first trimester) the uterine glands profiles appear in clusters. In late stage pregnancy (third trimester) the uterine gland profiles appear opposite chorionic vesicles. In the later stages of gestation maternal glandular epithelium consisted mainly of simple columnar epithelium. The cells had abundant flattened cisternae of granular endoplasmic reticulum usually with an apical-basal orientation. Their nuclei had abundant euchromatin relative to the amount of heterochromatin. They also had a prominent Golgi apparatus quite characteristic of protein synthesizing cells. The basal plasmalemma was thrown into infoldings that have the effect of increasing the surface area across which nutrients could pass from the maternal circulation and are secreted by the cell as histiotrophe. Both physiologic hypertrophy and hyperplasia of the uterine glands are observed to occur with advancement of pregnancy.

La histología y ultraestructura de las glándulas uterinas de la gálago menor (Galago senegalensis) fueron estudiadas en seis ejemplares (5 preñadas y 1 no preñada). Una parte de las glándula se fijó con Bouin y otra con glutaraldehído al 2,5% en tampón cacodilato 0,1 M. Una visión general de los principales resultados reveló que las glándulas uterinas en el útero no gestante son rudimentarias y escasas dentro de la mucosa. Al principio de la preñez (primer trimestre) las glándulas uterinas aparecen en racimos. En la última etapa de la preñez (tercer trimestre) las glándulas uterinas aparecen opuestas a las vesículas coriónicas. En las últimas etapas de gestación el epitelio glandular materno consiste principalmente en epitelio cilíndrico simple. Las células tenían abundantes cisternas aplanadas en el retículo endoplásmico rugoso, por lo general con una orientación apico-basal. Sus núcleos tenían abundante eucromatina en relación con la cantidad de heterocromatina. También tenían un aparato de Golgi prominente bastante característico de células que sintetizan proteínas. El plasmalema basal fue rechazado en repliegues que psoeen el efecto de aumentar el área de superficie a través del cual los nutrientes podrían pasar desde la circulación materna y son secretadas por las célula como histiotrofo. Hipertrofia fisiológica e hiperplasia de las glándulas uterinas se observaron con el avance de la preñez.

The estimated mechanical advantage of the prosimian ankle joint musculature, and implications for locomotor adaptation.

In this study we compared the power arm lengths and mechanical advantages attributed to 12 lower leg muscles across three prosimian species. The origins and insertions of the lower leg muscles in Garnett's galago, the ring-tailed lemur, and the slow loris were quantified and correlated with positional behaviour. The ankle joint of the galago has a speed-oriented mechanical system, in contrast to that of the slow loris, which exhibits more power-oriented mechanics. The lemur ankle joint exhibited intermediate power arm lengths and an intermediate mechanical advantage relative to the other primates. This result suggests that the mechanical differences in the ankle between the galago and the lemur, taxa that exhibit similar locomotory repertoires, reflect a difference in the kinematics and kinetics of leaping (i.e. generalised vs. specialised leapers). In contrast to leaping primates, lorises have developed a more power-oriented mechanical system as a foot adaptation for positional behaviours such as bridging or cantilevering in their arboreal habitat.

Projections of the superior colliculus to the pulvinar in prosimian galagos (Otolemur garnettii) and VGLUT2 staining of the visual pulvinar.

An understanding of the organization of the pulvinar complex in prosimian primates has been somewhat elusive due to the lack of clear architectonic divisions. In the current study we reveal features of the organization of the pulvinar complex in galagos by examining superior colliculus (SC) projections to this structure and comparing them with staining patterns of the vesicular glutamate transporter, VGLUT2. Cholera toxin subunit ß (CTB), Fluoro-ruby (FR), and wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) were placed in topographically different locations within the SC. Our results showed multiple topographically organized patterns of projections from the SC to several divisions of the pulvinar complex. At least two topographically distributed projections were found within the lateral region of the pulvinar complex, and two less obvious topographical projection patterns were found within the caudomedial region, in zones that stain darkly for VGLUT2. The results, considered in relation to recent observations in tree shrews and squirrels, suggest that parts of the organizational scheme of the pulvinar complex in primates are present in rodents and other mammals.

Morphological and neurochemical comparisons between pulvinar and V1 projections to V2.

The flow of visual information is clear at the earliest stages: the retina provides the driving (main signature) activity for the lateral geniculate nucleus (LGN), which in turn drives the primary visual cortex (V1). These driving pathways can be distinguished anatomically from other modulatory pathways that innervate LGN and V1. The path of visual information after V1, however, is less clear. There are two primary feedforward projections to the secondary visual cortex (V2), one from the lateral/inferior pulvinar and the other from V1. Because both lateral/inferior pulvinar and V2 cannot be driven visually following V1 removal, either or both of these inputs to V2 could be drivers. Retinogeniculate and geniculocortical projections are privileged over modulatory projections by their layer of termination, their bouton size, and the presence of vesicular glutamate transporter 2 (Vglut2) or parvalbumin (PV). It has been suggested that such properties might also distinguish drivers from modulators in extrastriate cortex. We tested this hypothesis by comparing lateral pulvinar to V2 and V1 to V2 projections with LGN to V1 projections. We found that V1 and lateral pulvinar projections to V2 are similar in that they target the same layers and lack PV. Projections from pulvinar to V2, however, bear a greater similarity to projections from LGN to V1 because of their larger boutons (measured at the same location in V2) and positive staining for Vglut2. These data lend support to the hypothesis that the pulvinar could act as a driver for V2.

Intrinsic signal optical imaging evidence for dorsal V3 in the prosimian galago (Otolemur garnettii).

Currently, we lack consensus regarding the organization along the anterior border of dorsomedial V2 in primates. Previous studies suggest that this region could be either the dorsomedial area, characterized by both an upper and a lower visual field representation, or the dorsal aspect of area V3, which only contains a lower visual field representation. We examined these proposals by using optical imaging of intrinsic signals to investigate this region in the prosimian galago (Otolemur garnettii). Galagos represent the prosimian radiation of surviving primates; cortical areas that bear strong resemblances across members of primates provide a strong argument for their early origin and conserved existence. Based on our mapping of horizontal and vertical meridian representations, visuotopy, and orientation preference, we find a clear lower field representation anterior to dorsal V2 but no evidence of any upper field representation. We also show statistical differences in orientation preference patches between V2 and V3. We additionally supplement our imaging results with electrode array data that reveal differences in the average spatial frequency preference, average temporal frequency preference, and sizes of the receptive fields between V1, V2, and V3. The lack of upper visual field representation along with the differences between the neighboring visual areas clearly distinguish the region anterior to dorsal V2 from earlier visual areas and argue against a DM that lies along the dorsomedial border of V2. We submit that the region of the cortex in question is the dorsal aspect of V3, thus strengthening the possibility that V3 is conserved among primates.

What is dental ecology?

Teeth have long been used as indicators of primate ecology. Early work focused on the links between dental morphology, diet, and behavior, with more recent years emphasizing dental wear, microstructure, development, and biogeochemistry, to understand primate ecology. Our study of Lemur catta at the Beza Mahafaly Special Reserve, Madagascar, has revealed an unusual pattern of severe tooth wear and frequent tooth loss, primarily the result of consuming a fallback food for which these primates are not dentally adapted. Interpreting these data was only possible by combining our areas of expertise (dental anatomy [FC] and primate ecology [MS]). By integrating theoretical, methodological, and applied aspects of both areas of research, we adopted the term "dental ecology"-defined as the broad study of how teeth respond to the environment. Specifically, we view dental ecology as an interpretive framework using teeth as a vehicle for understanding an organism's ecology, which builds upon earlier work, but creates a new synthesis of anatomy and ecology that is only possible with detailed knowledge of living primates. This framework includes (1) identifying patterns of dental pathology and tooth use-wear, within the context of feeding ecology, behavior, habitat variation, and anthropogenic change, (2) assessing ways in which dental development and biogeochemical signals can reflect habitat, environmental change and/or stress, and (3) how dental microstructure and macro-morphology are adapted to, and reflect feeding ecology. Here we define dental ecology, provide a short summary of the development of this perspective, and place our new work into this context.

Growth and the development of sexual size dimorphism in lorises and galagos.

Three fundamental ontogenetic pathways lead to the development of size differences between males and females. Males and females may grow at the same rate for different durations (bimaturism), grow for the same duration at different rates, or grow at a mix of rate and duration differences. While patterns of growth and the development of adult body size are well established for many haplorhines, the extent to which rate and duration differences affect strepsirrhine growth trajectories remains unclear. Here, we present iterative piecewise regression models that describe the ontogeny of adult body mass for males and females of five lorisoid species (i.e., lorises and galagos) from the Duke Lemur Center. We test the hypotheses that, like most haplorhines, sexual size dimorphism (SSD) is a result of bimaturism, and males and females of monomorphic species grow at the same rate for a similar duration. We confirm that the galagos in this sample (Galago moholi and Otolemur garnettii) show significant SSD that is achieved through bimaturism. Unlike monomorphic lemurids, the lorises in this sample show a diversity of ontogenetic patterns. Loris tardigradus does follow a lemur-like trajectory to monomorphism but Nycticebuscoucang and Nycticebus pygmaeus achieve larger adult female body sizes through a mixture of rate and duration differences. We show that contrary to previous assumptions, there are patterns of both similarity and difference in growth trajectories of comparably sized lorises and galagos. Furthermore, when ontogenetic profiles of lorisoid and lemurid growth are compared, it is evident that lorisoids grow faster for a shorter period of time.

Cortical projections to the superior colliculus in prosimian galagos (Otolemur garnetti).

The superior colliculus (SC) is a key structure within the extrageniculate pathway of visual information to cortex and is highly involved in visuomotor functions. Previous studies in anthropoid primates have shown that superficial layers of the SC receive direct inputs from various visual cortical areas such as V1, V2, and middle temporal (MT), while deeper layers receive direct inputs from visuomotor cortical areas within the posterior parietal cortex and the frontal eye fields. Very little is known, however, about the corticotectal projections in prosimian primates. In the current study we investigated the sources of cortical inputs to the SC in prosimian galagos (Otolemur garnetti) using retrograde anatomical tracers placed into the SC. The superficial layers of the SC in galagos received the majority of their inputs from early visual areas and visual areas within the MT complex. Yet, surprisingly, MT itself had relatively few corticotectal projections. Deeper layers of the SC received direct projections from visuomotor areas including the posterior parietal cortex and premotor cortex. However, relatively few corticotectal projections originated within the frontal eye fields. While prosimian galagos resemble other primates in having early visual areas project to the superficial layers of the SC, with higher visuomotor regions projecting to deeper layers, the results suggest that MT and frontal eye field projections to the SC were sparse in early primates, remained sparse in present-day prosimian primates, and became more pronounced in anthropoid primates.

Comparative morphological configuration of the cardiac nervous system in lorises and galagos (infraorder Lorisiformes, Strepsirrhini, primates) with evolutionary perspective.

The detailed systematic morphology and comparative anatomy of the autonomic cardiac nervous system (ACNS) with its surrounding structures were examined in 14 sides of 7 Lorisiformes species (lorises and galagos) to consider its evolutionary differences and similarities to others in the primate lineage. Our detailed descriptions clarified that (1) a family-dependent morphology of the ACNS does not exist although several variations are recognized as common variation in Lorisiformes, (2) the morphology of the ACNS in Lorisiformes is almost consistent regardless of the number of presacral vertebrae and cardiac positions, and (3) the ACNS morphology in Lorisiformes is extremely different from that in New and Old World monkeys, gibbons, and human, specifically in a narrower contribution of the superior cervical ganglion to the spinal cervical nerves, the caudal position and organization of the cervicothoracic ganglion, and the narrower origins of the sympathetic cardiac nerves, although all parasympathetic vagal cardiac branches are consistently present among the primates. Based on these detailed findings, the systematic and evolutionary morphologies of the ACNS are discussed and compared with our previous works. These results suggest the primate ACNS preserves its evolutionary history without modification and specialization by its functional adaptation as seen in the somatic system.

Cell-poor septa separate representations of digits in the ventroposterior nucleus of the thalamus in monkeys and prosimian galagos.

The architectonic features of the ventroposterior nucleus (VP) were visualized in coronal brain sections from two macaque monkeys, two owl monkeys, two squirrel monkeys, and three galagos that were processed for cytochrome oxidase, Nissl bodies, or the vesicular glutamate transporter 2 (vGluT2). The traditional ventroposterior medial (VPM) and ventroposterior lateral (VPL) subnuclei were easily identified, as well as the forelimb and hindlimb compartments of VPL, as they were separated by poorly staining, cell-poor septa. Septa also separated other cell groups within VPM and VPL, specifically in the medial compartment of VPL representing the hand (hand VPL). In one squirrel monkey and one galago we demonstrated that these five groups of cells represent digits 1-5 in a mediolateral sequence by injecting tracers into the cortical representation of single digits, defined by microelectrode recordings, and relating concentrations of labeled neurons to specific cell groups in hand VPL. The results establish the existence of septa that isolate the representation of the five digits in VPL of primates and demonstrate that the isolated cell groups represent digits 1-5 in a mediolateral sequence. The present results show that the septa are especially prominent in brain sections processed for vGluT2, which is expressed in the synaptic terminals of excitatory neurons in most nuclei of the brainstem and thalamus. As vGluT2 is expressed in the synaptic terminations from dorsal columns and trigeminal brainstem nuclei, the effectiveness of vGluT2 preparations in revealing septa in VP likely reflects a lack of synapses using glutamate in the septa.