Night and day: the comparative study of strepsirrhine primates reveals socioecological and phylogenetic patterns in olfactory signals.

Studies of chemical signals in vertebrates typically target single species; however, a broader understanding of olfactory communication may derive from comparative studies. We collected urine from 12 species representing most families of strepsirrhine primates--an excellent model clade because of variation in scent marking and socioecology. Using SPDE/GC-MS, we identified the volatile chemical composition of male and female urine from six 'urine marking' species and six glandular or 'non-urine marking' species. We found no sex differences, but as predicted, urine markers expressed the most chemically complex and distinctive urine. More distantly related species had more dissimilar urinary profiles, suggesting gradual signal evolution. Reconstructing ancestral chemical profiles revealed different evolutionary trajectories for urine and non-urine markers. We suggest that urine marking is an ancestral behaviour related to solitary, nocturnal living and that parallel evolutionary shifts towards greater reliance on derived glandular marking occurred in a family (Lemuridae) characterized by diurnality and sociality.

Honest olfactory ornamentation in a female-dominant primate.

Sexual selection theory predicts that potential mates or competitors signal their quality to conspecifics. Whereas evidence of honest visual or vocal signals in males abounds, evidence of honest signalling via scent or by females is scarce. We previously showed that scent marks in male lemurs seasonally encode information about individual heterozygosity - a reliable predictor of immunocompetence and survivorship. As female lemurs dominate males, compete over resources, and produce sexually differentiated scent marks that likely evolved via direct selection, here we tested whether females also advertise genetic quality via olfactory cues. During the breeding season specifically, individual heterozygosity correlated negatively with the diversity of fatty acids (FAs) expressed in labial secretions and positively with the diversity of heavy FA esters. As odour-gene relationships predictive of health and survivorship emerged during a period critical to mate choice and female competition, we posit that genital scent marks function as honest olfactory ornaments in females.

Dietary vitamins A and E influence retinyl ester composition and content of the retinal pigment epithelium.

Experiments were conducted to determine the influence of dietary levels of vitamin A and alpha-tocopherol on the amounts and composition of retinyl esters in the retinal pigment epithelium of light-adapted albino rats. Groups of rats were fed diets containing alpha-tocopherol and either no retinyl palmitate, adequate retinyl palmitate, or excessive retinyl palmitate. Other groups of rats received diets lacking alpha-tocopherol and containing the same three levels of retinyl palmitate. Retinoic acid was added to diets lacking retinyl palmitate. After 27 weeks, the animals were light-adapted to achieve essentially total visual pigment bleaches, and the neural retinas and retinal pigment epithelium-eyecups were then dissected from each eye for vitamin A ester determinations. Almost all of the retinyl esters were found in the retinal pigment epithelium-eyecup portions of the eyes, mainly as retinyl palmitate and retinyl stearate. Maintaining rats on a vitamin A-deficient, retinoic acid-containing diet led to significant reductions in retinal pigment epithelial retinyl ester levels in rats fed both the vitamin E-supplemented and vitamin E-deficient diets; contrary to expectations, the effect of dietary vitamin A deficiency was more pronounced in the vitamin E-supplemented rats. Vitamin A deficiency in retinoic acid-maintained animals also led to significant reductions in retinyl palmitate-to-stearate ester ratios in the retinal pigment epithelia of both vitamin E-supplemented and vitamin E-deficient rats. Excessive dietary intake of vitamin A had little, if any, effect on retinal pigment epithelial retinyl ester content or composition. Vitamin E deficiency resulted in significant increases in retinal pigment epithelial retinyl palmitate content and in palmitate-to-stearate ester ratios in rats fed all three levels of vitamin A, but had little effect on retinal pigment epithelial retinyl stearate content. In other tissues, vitamin E deficiency has been shown to lower vitamin A levels, and it is widely accepted that this effect is due to autoxidative destruction of vitamin A. The increase in retinal pigment epithelial vitamin A ester levels in response to vitamin E deficiency indicates that vitamin E does not regulate vitamin A levels in this tissue primarily by acting as an antioxidant, but rather may act as an inhibitor of vitamin A uptake and/or storage. The effect of vitamin E on pigment epithelial vitamin A levels may be mediated by the vitamin E-induced change in retinyl palmitate-to-stearate ratios.

Relationship between dietary retinol and lipofuscin in the retinal pigment epithelium.

A variety of evidence suggests that autoxidation of cellular components probably plays a significant role in the age-related accumulation of lipofuscin, or age-pigment, in the mammalian retinal pigment epithelium (RPE). Among the likely candidates for conversion into RPE lipofuscin fluorophores via autoxidative mechanisms are vitamin A compounds, which are present in the retina and RPE in high concentrations. Vitamin E, an important lipid antioxidant, is likely to inhibit vitamin A autoxidation. Experiments were conducted to evaluate the significance of vitamin A autoxidation in the deposition of lipofuscin in the RPE. Albino rats were fed diets either supplemented with or lacking vitamin E. Each of these two groups of animals was further subdivided into three groups which were fed different levels of vitamin A palmitate: none, 14.0 mumol/kg diet, and 80.5 mumol/kg diet. After 26 weeks, the animals were killed and the RPE lipofuscin contents were determined by both fluorescence measurements and quantitative ultrastructural morphometry. Vitamin A palmitate deficiency led to significant reductions in RPE lipofuscin deposition, relative to the amounts of this pigment present in the groups receiving vitamin A palmitate in their diets. The relative magnitude of the vitamin A effect was greater in the vitamin E-supplemented groups than in the groups fed the diets deficient in vitamin E. This finding suggests that vitamin E interacts with vitamin A ester metabolites in vivo in a more complex manner than simply acting as an antioxidant protectant. Rats fed the diets containing the higher level of vitamin A palmitate failed to display elevated RPE lipofuscin contents relative to those in the rats fed 14.0 mumol of vitamin A palmitate/kg diet. Failure of high vitamin A intake to enhance RPE lipofuscin deposition may have been due to the fact that intake of vitamin A above normal levels did not lead to an elevation in vitamin A content of the retinal tissue. Establishing an effect of vitamin A deficiency on RPE lipofuscin deposition and characterization of the interactions between vitamins E and A are important steps toward defining precisely the molecular and cellular mechanisms underlying age-pigment accumulation in the RPE.

Exposure to naturally circulating androgens during foetal life incurs direct reproductive costs in female spotted hyenas, but is prerequisite for male mating.

Among all extant mammals, only the female spotted hyena (Crocuta crocuta) mates and gives birth through the tip of a peniform clitoris. Clitoral morphology is modulated by foetal exposure to endogenous, maternal androgens. First births through this organ are prolonged and remarkably difficult, often causing death in neonates. Additionally, mating poses a mechanical challenge for males, as they must reach an anterior position on the female's abdomen and then achieve entry at the site of the retracted clitoris. Here, we report that interfering with the actions of androgens prenatally permanently modifies hyena urogenital anatomy, facilitating subsequent parturition in nulliparous females who, thereby, produce live cubs. By contrast, comparable, permanent anatomical changes in males probably preclude reproduction, as exposure to prenatal anti-androgens produces a penis that is too short and has the wrong shape necessary for insertion during copulation. These data demonstrate that the reproductive costs of clitoral delivery result from exposure of the female foetus to naturally circulating androgens. Moreover, the same androgens that render an extremely unusual and laborious process even more reproductively costly in the female are apparently essential to the male's physical ability to reproduce with a normally masculinized female.

Neonatal testosterone and handedness in yearling rhesus monkeys (Macaca mulatta).

This study investigated the relationship between neonatal testosterone (T) and hand bias in young rhesus monkeys (Macaca mulatta). Subjects (n = 8 per group) included: neonatally androgen-suppressed males, using a Nal-Lys gonadotropin releasing hormone (GnRH) antagonist (Antide); androgen-suppressed males receiving T replacement by a long-acting T preparation (CDB); control males; and control females. Antide suppressed T to the female range, whereas CDB replacement produced supranormal levels. Visually guided reaching, in a social context, showed a population-level left-hand bias. Males with elevated T did not show a stronger left-hand bias than males with normal T, but did show a stronger bias for the preferred hand whether left or right. Males with Antide-suppressed T showed an intermediate degree of hand bias. Results suggest that high neonatal T levels affect laterality and raise the possibility that GnRH analogues influence brain development. These data suggest a broad influence of the CNS-pituitary-testicular axis on brain asymmetries and provide support for an early neonatal period of T-influenced brain differentiation.

Status, age, and sex effects on performance of discrimination tasks in group-tested rhesus monkeys (Macaca mulatta).

To assess the relation between performance and social or demographic variables, this study group tested a captive monkey colony on visual and manual discrimination problems. Animals could choose between differently colored, sand-filled boxes, where hue signaled the initial probability of finding buried food items. Dominant animals and subadults were most successful in locating and retrieving incentives, but sex did not affect performance. Rank effects occurred without overt aggression, suggesting deference by subordinates as a mediating mechanism. Age effects may reflect changing attention patterns only evident in complex arenas where cue salience becomes diluted. Because these findings differ from studies of singly tested animals, they show that, in a social context, an individual's rank and age may define opportunities to gain or efficiently use information.

Asymmetrical hand use in rhesus monkeys (Macaca mulatta) in tactually and visually regulated tasks.

Asymmetrical hand use by rhesus monkeys (Macaca mulatta) was investigated in a series of tactually and visually guided tasks. The 1st experiment recorded manual preferences of 29 monkeys for solving a haptic discrimination task in a hanging posture. There was a left-hand population bias: 21 monkeys had a left-hand bias, 4 a right-hand bias, and 4 no bias. The 2nd experiment, 4 tasks with 23 to 51 monkeys, investigated the critical components of the 1st experiment by varying the posture (hanging, sitting, or tripedal) and the sensory requirements (tactile or visual). Posture influenced hand bias, with a population-level left-hand bias in hanging and sitting postures, but an almost symmetrical distribution in the tripedal posture. A left-hand bias was found for both sensory modalities, but the bias was stronger in the tactual tasks. Results suggest a possible right-hemisphere specialization in the rhesus for tactile, visual, or spatial processing.

Social context affects how rhesus monkeys explore their environment.

This study reports on social modulation of exploratory behavior and response to novelty by members of a captive rhesus monkey colony. The group was trained to split in half, with one subgroup composed of dominant members only, the other of subordinates. The animals were then presented the same initially novel stimuli (i.e., sand-filled metal boxes containing hidden food items) in two social contexts differing in hierarchical composition. In a combined context, all group members (i.e., both subgroups together) were simultaneously presented the stimuli. In a split context, only members of the top or bottom half of the group (i.e., each subgroup in turn) was independently presented the stimuli. Subordinates responded similarly to dominant animals in the combined context but differently in the split context, where they were far more hesitant. Rank-related differences were evident in the way animals used their home compound and in their approach and responsiveness toward the stimuli. These findings show that social context influences how animals explore novel situations, possibly reflecting different social roles or status effects on the perception of social structure. Also, despite the complexity of primate social relationships, the separation technique produced no permanent or adverse effects on the social integrity of the group. This study shows that manipulating the social environment through separation training can be a powerful tool for assessing contextual influences on behavior.

Low-status monkeys "play dumb" when learning in mixed social groups.

Many primates, including humans, live in complex hierarchical societies where social context and status affect daily life. Nevertheless, primate learning studies typically test single animals in limited laboratory settings where the important effects of social interactions and relationships cannot be studied. To investigate the impact of sociality on associative learning, we compared the individual performances of group-tested rhesus monkeys (Macaca mulatta) across various social contexts. We used a traditional discrimination paradigm that measures an animal's ability to form associations between cues and the obtaining of food in choice situations; but we adapted the task for group testing. After training a 55-member colony to separate on command into two subgroups, composed of either high- or low-status families, we exposed animals to two color discrimination problems, one with all monkeys present (combined condition), the other in their "dominant" and "subordinate" cohorts (split condition). Next, we manipulated learning history by testing animals on the same problems, but with the social contexts reversed. Monkeys from dominant families excelled in all conditions, but subordinates performed well in the split condition only, regardless of learning history. Subordinate animals had learned the associations, but expressed their knowledge only when segregated from higher-ranking animals. Because aggressive behavior was rare, performance deficits probably reflected voluntary inhibition. This experimental evidence of rank-related, social modulation of performance calls for greater consideration of social factors when assessing learning and may also have relevance for the evaluation of human scholastic achievement.

Age-related alterations in vitamin A metabolism in the rat retina.

Vitamin A plays a central role in visual transduction and in maintaining the structural integrity of the retina. It is possible that age-related alterations in vitamin A metabolism in the eye could contribute to the impairment of visual function that occurs during senescence. Therefore, investigations were conducted to determine whether the metabolism of this vitamin in the rat retina was altered during aging. Pigmented rats aged 12-, 22-, and 32 months were dark-adapted, and one eye from each animal was enucleated under dim red light. The neural retinas were separated from the retinal pigment epithelium (RPE)-choroid-scleral complexes, and the amounts and forms of vitamin A in both tissues were determined. The animals were then fully light-adapted, and the same measurements were performed on the tissues from the remaining eye of each rat. A number of age-related alterations in the vitamin A composition and content of the retina and RPE were observed. The most pronounced of these changes were significant increases in the ratios of retinyl palmitate to retinyl stearate with advancing age in both the neural retina and RPE. The total vitamin A ester contents of the RPEs increased during senescence in the dark-adapted state, but not in the light-adapted state. Retinyl ester levels in the neural retinas, on the other hand, did not differ significantly between 12- and 32-month-old animals in either the light-adapted or dark-adapted states. The amounts of all-trans retinol in the neural retinas decreased during aging, mainly in the dark-adapted state, whereas aging had no influence on RPE all-trans retinol content. The age-related alterations in metabolism of vitamin A that these observations reflect may be related to certain changes in visual function that occur during senescence.

Influence of early photoreceptor degeneration on lipofuscin in the retinal pigment epithelium.

Experiments were conducted to evaluate the role played by photoreceptor cells in the accumulation of age pigment, or lipofuscin, in the retinal pigment epithelium (RPE). The age-related accumulation of RPE lipofuscin was compared between rats with hereditary photoreceptor degeneration (RDY) and congenic rats with normal retinas. In the RDY animals, the age-related increase in RPE lipofuscin content was substantially less than in normal controls. This suggests that the photoreceptor cells play a significant role in RPE lipofuscin deposition, although they may not be the sole contributors to RPE lipofuscin formation. Evidence that outer-segment components may be converted into lipofuscin fluorophores was provided by the discovery that in young RDY rats, fragments of outer segments from degenerating photoreceptor cells had fluorescence properties similar to those of RPE lipofuscin. Chloroform-methanol extraction of retina-RPE tissue from young normal and dystrophic rats, and analysis of the chloroform fractions by thin-layer chromatography, revealed three distinct fluorescent components associated with the lipofuscin-like fluorescence of the outer-segment fragments in the RDY rats.

Polyol and vacuole formation in cultured canine lens epithelial cells.

Polyol accumulation and myo-inositol depletion were accompanied by extensive vacuole formation in cultured canine lens epithelial cells that were incubated for up to 96 hr in growth medium supplemented with 30 mM D-galactose or 30 mM D-glucose. These changes did not occur in cells incubated in a hypergalactosemic or hyperglycemic medium which also contained an aldose reductase inhibitor (20 microM sorbinil). In addition, these changes were not observed in lens cells incubated in growth medium supplemented with either 30 mM mannitol, which is known to enter cells only slowly, or in 30 mM L-galactose, which is not a substrate for aldose reductase. The vacuoles were visible at the ultrastructural level after 6 hr of incubation in 30 mM D-galactose and increased in both number and size with time. These vacuoles had a unique fine structure. They did not result from swelling of mitochondria or other cell organelles. As demonstrated cytochemically, they did not represent either lysosomes or Golgi saccules. The proliferation pattern of cells incubated with 30 mM D-galactose was clearly different from that of control cells, but approached normal when an aldose reductase inhibitor was added to the incubation medium. Together these findings suggest that vacuole formation and altered cell proliferation were caused by polyol accumulation and/or myo-inositol loss, both of which result from aldose reductase activity.

Androgens and masculinization of genitalia in the spotted hyaena (Crocuta crocuta). 3. Effects of juvenile gonadectomy.

Studies involving the administration of anti-androgens to spotted hyaenas during fetal development have raised questions concerning the precise contributions of steroids to phallic growth in these animals. If gonadal androgens promote postnatal penile growth in males, the following would be expected: (a) a period of accelerated growth accompanying achievement of puberty, and (b) a marked reduction in adult penile size and density of penile spines after gonadectomy. If a similar androgenic pubertal process stimulates clitoral growth in these highly 'masculinized' hyaenas, parallel observations in females would be expected; however, the role of oestrogens in accounting for female-typical clitoral development would also have to be considered. The results of the present study suggest a limited role, if any for androgenic stimulation of phallic growth. That is, penile growth was greater during the 10 month period preceding puberty, than during an 18-month period that included the traditional increase in pubertal androgens. In addition, pre-pubertal castration had minimal effects on penile length, diameter, or the presence of penile spines. In females, most clitoral growth also occurred before puberty, although pre-pubertal ovariectomy produced significant reductions in clitoral diameter and the elasticity of the urogenital meatus. These feminine characteristics which normally distinguish the female from the male phallus in this species, were partially restored by a brief period of oestrogen administration. Both sexes displayed erections many years after pre-pubertal castration. The results of the present study suggest that postnatal phallic growth is largely independent of gonadal steroids, with oestrogenic facilitation of female-typical clitoral characteristics in spotted hyaenas.

Androgens and masculinization of genitalia in the spotted hyaena (Crocuta crocuta). 2. Effects of prenatal anti-androgens.

Pregnant spotted hyaenas were treated with anti-androgens to interfere with the unusually masculine 'phallic' development that characterizes females of this species. The effects on genital morphology and plasma androgen concentrations of infants were studied during the first 6 months of life. Although there were consistent 'feminizing' effects of prenatal anti-androgen treatment on genital morphology in both sexes, such exposure did not produce males with extreme hypospadia, as it does in other species, nor did it produce females with a 'typical' mammalian clitoris and external vagina. 'Feminization' of males resulted in a penis with the morphological features of the hyaena clitoris, and 'feminization' of females exaggerated the sex differences that are typical of this species. The effects of treatment were present at birth and persisted for at least 6 months. Treatment of pregnant females with flutamide and finasteride also markedly reduced circulating concentrations of testosterone and dihydrotestosterone in maternal plasma during pregnancy. Plasma delta 4-androstenedione was reduced in the female, but not the male, infants of treated mothers, consistent with an epigenetic hypothesis previously advanced to explain hormonal 'masculinization' of females. The present 'feminizing' effects of prenatal anti-androgen treatment are consistent with contemporary understanding of sexual differentiation, which accounts for morphological variation between the sexes in terms of steroids. However, current theory does not account for the basic genital structure of females and the present data suggest that development of the male penis and scrotum, and the female clitoris and pseudoscrotum, in spotted hyaenas may involve both androgen-dependent and androgen-independent components.