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.

Female reproductive activity and its endocrine correlates in the African lesser bushbaby, Galago moholi.

Steroid hormones play an important role in female reproductive physiology and behaviour and are often used to monitor important female reproductive events. However, such studies are often attempted on captive populations alone, delivering limited data. One such example is the African lesser bushbaby, Galago moholi, for which contradicting observational data exist between captive and free-ranging populations, while hormonal analyses have only been obtained from a single captive population. To extend and rectify the limited information, we monitored faecal progestagen and oestrogen metabolite levels across various important life history stages of both captive and free-ranging G. moholi. We additionally recorded changes in vaginal state as well as the occurrence of reproductive and aggressive behaviour throughout the study. Data from our captive population revealed an ovarian cycle length of 33.44 ± 0.59 days (mean ± SD), with follicular and luteal phases of 14.2 ± 1.0 and 19.1 ± 1.5 days, respectively, and an average pregnancy length of 128 ± 3.3 days. The initiation of female reproductive activity was closely linked to an oestrus-related increase in faecal oestrogen metabolite levels. Four of the seven captive females monitored in our study conceived during the May mating period, with one additional female fertilised in September, supporting the idea that the September mating period functions as a back-up for female G. moholi. Identified benchmark faecal progestagen metabolite levels (non-pregnant: >1 µg/g dry weight (DW), pregnant: >9 µg/g DW) should help researchers to determine pregnancy status of randomly wild-caught females in even a cross-sectional study setup.

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.

Identification of a new genomic hot spot of evolutionary diversification of protein function.

Establishment of phylogenetic relationships remains a challenging task because it is based on computational analysis of genomic hot spots that display species-specific sequence variations. Here, we identify a species-specific thymine-to-guanine sequence variation in the Glrb gene which gives rise to species-specific splice donor sites in the Glrb genes of mouse and bushbaby. The resulting splice insert in the receptor for the inhibitory neurotransmitter glycine (GlyR) conveys synaptic receptor clustering and specific association with a particular synaptic plasticity-related splice variant of the postsynaptic scaffold protein gephyrin. This study identifies a new genomic hot spot which contributes to phylogenetic diversification of protein function and advances our understanding of phylogenetic relationships.

Differential expression patterns of striate cortex-enriched genes among Old World, New World, and prosimian primates.

A group of 5 genes, OCC1, testican-1, testican-2, 5-HT1B, and 5-HT2A, are selectively expressed in layer 4 (4C of Brodmann) of striate cortex (visual area V1) of both Old World macaques and New World marmoset monkeys. The expression of these genes is activity dependent, as expression is reduced after blocking retinal activity. Surprisingly, the pronounced expression pattern has not been found in rodents or carnivores. Thus, these genes may be highly expressed in V1 of some but perhaps not all primates. Here, we compared the gene expression in members of 3 major branches of primate evolution: prosimians, New World monkeys, and Old World monkeys. Although the expression pattern of 5-HT1B was well conserved, those of the other genes varied from the least distinct in prosimian galagos to successively more in New World owl monkeys, marmosets, squirrel monkeys, and Old World macaque monkeys. In owl monkeys, the expression of 5-HT2A was significantly reduced by monocular tetrodotoxin injection, while those of OCC1 and 5-HT1B were not. Thus, we propose that early primates had low levels of expression and higher levels emerged with anthropoid primates and became further enhanced in the Old World catarrhine monkeys that are more closely related to humans.

The development of small primate models for aging research.

Nonhuman primate (NHP) aging research has traditionally relied mainly on the rhesus macaque. But the long lifespan, low reproductive rate, and relatively large body size of macaques and related Old World monkeys make them less than ideal models for aging research. Manifold advantages would attend the use of smaller, more rapidly developing, shorter-lived NHP species in aging studies, not the least of which are lower cost and the ability to do shorter research projects. Arbitrarily defining "small" primates as those weighing less than 500 g, we assess small, relatively short-lived species among the prosimians and callitrichids for suitability as models for human aging research. Using the criteria of availability, knowledge about (and ease of) maintenance, the possibility of genetic manipulation (a hallmark of 21st century biology), and similarities to humans in the physiology of age-related changes, we suggest three species--two prosimians (Microcebus murinus and Galago senegalensis) and one New World monkey (Callithrix jacchus)--that deserve scrutiny for development as major NHP models for aging studies. We discuss one other New World monkey group, Cebus spp., that might also be an effective NHP model of aging as these species are longer-lived for their body size than any primate except humans.

Distribution of calcium-binding proteins within the parallel visual pathways of a primate (Galago crassicaudatus).

Bush babies possess three distinct parallel pathways to striate cortex (V1 or area 17). The calcium-binding proteins parvalbumin (PV) and calbindin (CB) typically show complementary regional distributions in the brain, often associated with specific aspects of functionally related groups of cells. We asked whether PV+ and CB+ immunoreactivity differentiate central visual parallel pathways in this species. Results show that PV and CB cell and neuropil staining is strongly complementary in the lateral geniculate nucleus (LGN) and is associated with separate parallel pathways. CB+ immunoreactivity is dense, but cytochrome oxidase (CO) staining is light in the paired koniocellular layers. PV+ and CO+ immunoreactivity is most dense in the parvocellular and magnocellular layers. Combined analyses of cell size, retrograde labeling, and double labeling have confirmed that all PV+ and CB+ LGN cells are geniculocortical relay cells; none was found to be gamma-aminobutyric acid (GABA)ergic. In V1, dense PV+ neuropil closely matches the expression of CO in layer 4 and in the blobs of layer 3. CB+ staining is most dense in layers 2 and 3A and is not strongly expressed within the CO interblobs. Finally, PV and CB are not found in related parallel pathway components in the LGN and V1 (e.g., in V1, CO blobs exhibit dense PV+ neuropil, yet they are targets of the small K geniculocortical relay cells that are CB+ in the LGN). Our findings support the view that three functionally distinct visual pathways project to V1 from the LGN. However, the differences in the patterns of localization of PV and CB in the LGN and in V1 suggest that these proteins may be utilized in different ways in these two visual areas.

Hematology, serum chemistry values, and rectal temperatures of adult greater galagos (Galago garnetti and G. crassicaudatus).

Hematological and serum chemistry values, as well as rectal temperatures, were obtained from greater galagos (Galago garnettii and G. crassicaudatus), in order to establish normative values. No species or sex differences were found for four hematological parameters and 15 serum chemistry parameters. Species differences were seen in phosphate, magnesium, cholesterol, alkaline phosphate, G-glutamyl transferase, mean corpuscular volume and leucocyte, neutrophil, and lymphocyte number. Significant sex differences were observed in glucose, hemoglobin, and hematocrit values. Species and sex differences were seen in chloride and erythrocyte number.

Organization of cytochrome oxidase staining in the visual cortex of nocturnal primates (Galago crassicaudatus and Galago senegalensis): I. Adult patterns.

The distribution and differential staining patterns of cytochrome oxidase (CO) activity in visual cortical areas have provided useful anatomical markers for the modular organization of area 17 (striate cortex) and area 18 in primates. In macaque and squirrel monkeys, previous studies have shown that the majority of cells that lie in areas of high CO activity are color selective, are nonoriented, and project to adjacent zones of high CO activity in area 17 and to stripes of high CO activity in area 18. By contrast, most cells in zones with weak CO activity in area 17 have relatively narrow orientation tuning and are not color selective (Livingstone and Hubel: J. Neurosci. 4:309-356, 2830-2835, '84; 7:3371-3377, '87). The periodic organization of CO activity in area 17, the "blobs," and the stripe-like organization in area 18 thus seem to define visual cortical processing modules and/or channels in primates. We have investigated the organization of CO activity in areas 17 and 18 in two species of nocturnal prosimian primates [Galago crassicaudatus (GCC) and Galago senegalensis (GSS)] in order to evaluate CO staining patterns in primates that have been reported to possess almost exclusively rod retinae and no color vision. In area 17 of both species, our results show that, as in diurnal and nocturnal simian primates, the darkest CO staining occurs in layers III and IV, with clear periodicity in layer III (i.e., CO blobs) and homogeneous staining in layer IV beta, the cortical recipient sublayer of the geniculate parvocellular layers. In GCC, individual blobs in layer III appear to be larger and less frequent than has been reported for the macaque monkey. Unlike simian primates, both galago species exhibit clear CO periodicities within layer IV alpha, the cortical recipient sublayer of the magnocellular geniculate layers. In addition, faint CO periodicities are apparent in layer VI and scattered large darkly CO stained pyramidal cells are visible throughout layer V. Quantitative analysis suggests that CO periodicities are more frequent in GSS than in GCC, suggesting that there may be evolutionary pressure to maintain the same number of CO modules within the smaller striate cortex of the lesser galago, although this is not the trend found across distantly related species. CO activity in area 18 is less well-developed than reported in other primates. In fact, we could not reliably identify discontinuities in CO staining in area 18 of GSS.(ABSTRACT TRUNCATED AT 400 WORDS)

Scaling of growth and life history traits relative to body size, brain size, and metabolic rate in lorises and galagos (Lorisidae, primates).

A broad range of variation in body size, brain size, and metabolic rate occurs within the primate family Lorisidae, thus providing an opportunity to examine the relationship of these three parameters to variation in growth and life history traits. Data on adult body weight, gestation length, lactation length, age at first estrus, litter size, and growth parameters were collected from a captive colony of four lorisid species, Loris tardigradus, Nycticebus coucang, Galago crassicaudatus, and G. senegalensis. The data presented here constitute the most complete life history information available for these poorly understood prosimian species. Correlation and allometric analyses were performed to determine the relationships between variables. Among the lorisids studied, adult body weight, adult cranial capacity, and relative cranial capacity did not predict variation in life history traits. Adult basal metabolic rate predicted most of the variability in gestation length, lactation length, and growth parameters. Lorisines differ from similarly sized galagines in having lower basal metabolic rates, slower growth rates, slower developmental rates, and smaller litter sizes, resulting in reduced reproductive potential. This may be a consequence of lorisine adaptation to a diet of toxic insects. Metabolic rate and diet may be among the most important parameters to examine in any study of life history evolution.

Glutamic acid decarboxylase-immunoreactive neurons and horseradish peroxidase-labeled projection neurons in the ventral posterior nucleus of the cat and Galago senegalensis.

Immunocytochemical methods were used to identify neurons in the ventral posterior nucleus of the cat and Galago senegalensis that contain glutamic acid decarboxylase (GAD), the synthetic enzyme for the inhibitory neurotransmitter, GABA. In both species GAD-immunoreactive neurons make up about 30% of the total neurons in the ventral posterior nucleus and form a distinct class of small cells. After cortical injections of horseradish peroxidase (HRP), GAD-immunoreactive cells are not labeled with HRP and may, therefore, be GABAergic local circuit neurons. Comparison of the dendritic morphology of GAD-immunoreactive neurons with that of HRP-filled projection neurons reveals that the morphology of the GAD-containing neurons is distinct and, in particular, that the GAD-immunoreactive neurons display fewer primary dendrites. The relay neurons, in turn, can be divided into classes based on dendritic morphology and cell body size.

Progestagen-concentrating cells in the brain, uterus, vagina and mammary glands of the galago (Galago senegalensis).

Progestagen-concentrating cells were localized in the oestrogen-primed ovariectomized galago by radioautography after injection of [3H]promegestone (R5020). In the brain, radioactivity was concentrated in the nuclei of neurones in the preoptic region and in the mediobasal hypothalamus. Labelled cells were also observed in the anterior pituitary. In the uterus (uterine horns and cervix), the muscle and stromal cells showed greater labelling than did the glandular and luminal epithelia. Labelled cells were present in the different cell layers of the vagina. The majority of glandular epithelial cells of the mammary glands exhibited a high degree of labelling. Pretreatment with an excess of unlabelled promegestone but not with an excess of nonradioactive testosterone reduced the nuclear concentration of radioactivity in these target tissues. These results show that there are no major differences in the distribution of progestagen-concentrating cells in rodents and galago.

Immunofluorescence study of LRH-producing neurons in prosimians (Tupaia and Galago).

Reactive LRH neurons were characterized in prosimians (Tupaia and Galago) by immunofluorescence using rabbit immunesera against unconjugated synthetic LRH, or LRH conjugated with bovine serum albumin. These neurons, which vary individually in number in one species, are mainly concentrated in the rostral hypothalamus (medial preoptic area and anterior hypothalamic area) and in the lamina terminalis. In contrast to the simians and man, immunoreactive perikarya were not routinely found in the mediobasal hypothalamus of the prosimians investigated in the present study. Reactive axons of the hypothalamo-hypophyseal tract are more numerous and conspicuous in the retrochiasmatic area and in the postinfundibular eminence. They give rise to radiating collaterals ending mainly around the capillaries of the primary portal plexus of the median eminence and of the infundibular stem (where they are generally more numerous). Reactive axons of the preoptico-terminal tract, originating from the perikarya of the lamina terminalis, end around the capillaries of the vascular organ or below and between the ependymal cells lining its ventricular side. In Galago a small but very distinct tract of reactive axons runs under the optic chiasma, between the lamina terminalis and the ventral labium of the infundibulum. Very fine reactive extrahypothalamic axons were observed in the posterior part of the habenular ganglia, along the preamygdaloid portion of the stria terminalis and along the blood vessels of the parolfactory area.