Scent,sex, and the self-calibrating rat

We propose that one mechanism whereby male rats,Rattus norvegicus, might gauge the reproductive condition of a female is by calibrating a stable odor with an odor that fluctuates through her reproductive cycle. We provide behavioral and histological evidence in support of such a self-calibration model. Male rats sniffed frequently at various body zones of females, and the proportion of sniffs deployed to each zone varied with the females' reproductive condition and relatedness. The females' haunches received more sniffs than any other part of their bodies, irrespective of their relatedness or reproductive condition. Furthermore, males tended to sniff the haunch after sniffing the forequarters, as part of a sequence of sniffing along a female from forequarters to hindquarters. Histology of the skin sebaceous glands indicated that the secretory activity of glands in the haunch, but not those in the forequarters, changed during estrus. Therefore, male rats had the opportunity to judge a female's reproductive status by calibrating the odor of her haunch against that of her forequarters. Self-calibration could represent a means of accommodating differences in odor within individuals.     

Rates of urine excretion by house mouse (Mus domesticus): Differences by age,sex, social status,and reproductive condition

A series of five experiments was used to test hypotheses about factors affecting excreted urine output per day in male and female house mice (Mus domesticus). Urine was collected in metabolism cages over a 24-hr period. Male house mice excrete urine at a rate 1.5–2.0 times that of females. Daily average urine output increases with age for both sexes and for mice of the same age; urine output per day is correlated with body mass. Females in estrus produce more urine than females in diestrus. Urine output per day increases during the latter two thirds of pregnancy and remains high throughout lactation. Density does not influence urine output per day for either sex over the range of densities tested. Castration reduces urine output per day in male mice, but ovariectomy in females does not alter rates of urine production. Dominant males produce more urine than subordinate males, but there are no similar effects for female mice. The findings have potential implications for our understanding of the functional and ecological uses of urine by house mice.     

Morphology and chemistry of brown rat,Rattus norvegicus,preputial and clitoral glands

In contrast to previous results from laboratory rats, when sexual dimorphism occurred, clitoral glands from wild female rats were heavier and accounted for a greater proportion of body mass than preputial glands from wild males. Gland length and weight increased with body weight, but gland weight as a proportion of body weight did not. Preputial and clitoral glands showed seasonal differences in size: at one site glands were smaller in December than in April. Maturity, estrus, and pregnancy had no significant effect on clitoral gland size, but immature males had proportionally smaller preputial glands than mature males, and the absolute gland weight, but not gland weight as a proportion of body weight, increased with testes weight. There were no differences between the sexes in lipid production, which increased with preputial or clitoral gland weight. An homologous series of aliphatic acids and their methyl and ethyl esters, plus squalene, cholesterol and lanosterol, was revealed by GC-MS study of the trimethyl silyl derivatized lipid extract. Males and females could be distinguished by principal component analysis of chromatogram peaks.     

Stimulation of Estrus in Female Mice by Male Urinary Proteins

Stimulation of estrus in adult female mice was obtained with major urinary proteins (MUPs) with the natural volatile ligands bound. The MUP threshold concentration for this effect was about 1.8 mg/ml. MUPs without the ligands bound, as purified by organic extraction of hydrophobic compounds, stimulated estrus in mice only when dissolved in carrier urine of juvenile or castrated adult male mouse or ovariectomized female mouse. They did not stimulate estrus when dissolved in water. Mice that had the vomeronasal organ removed were insensitive to MUPs. It is concluded that MUPs are an integral part of the mouse male pheromones that stimulate hormonal activity in females and that the vomeronasal system is involved in the estrus-stimulating effect of the major urinary proteins.     

Foveal glands,source of sex pheromone production in the ixodid tickDermacentor andersoni stiles

The foveal glands of the Rocky Mountain wood tick,Dermacentor andersoni Stiles, are the sex pheromone glands from which the sex pheromone is released via the foveae dorsales. The sex pheromone, 2,6-dichlorophenol, was recovered from extracts of these glands by GLC. Other evidence of the role of these glands in sex pheromone production is described. A³⁶Cl-labelled volatile compound (or compounds) was (were) collected from partially engorged femaleD. andersoni fed in³⁶Cl-labelled hosts, but no labelled compounds were collected when the foveae dorsales were blocked. X-ray analysis revealed unusual concentrations of chlorine in the foveal glands compared to other tissues. Autoradiography also revealed significant accumulations of radiochlorine in the vicinity of these glands.Presumably, the foveal glands of the American dog tick,Dermacentor variabilis (Say), are the sex pheromone glands of that species also, since a³⁶Cl-labelled volatile was collected from female ticks fed on a³⁶Cl-labelled host. However, attempts to recover 2,6-dichlorophenol from gland extracts or volatile emissions fromD. variabilis were unsuccessful.Supported by grants AI 10,986 and AI 10,987 from the National Institute of Allergy and Infectious Diseases, U.S. Public Health Service, U.S. Department of Health, Education and Welfare, Bethesda, Maryland, 20014.     

Evidence for the Site of Female Sex Pheromone Production in Periplaneta americana

We used bioassays to determine the sex pheromone production site in the female American cockroach (Periplaneta americana). Bioassayed extracts from various body parts aroused sexual responses from unmated males. However, the extract of hindgut (including both colon and rectum) induced strong responses. Colon extract stimulated the strongest sexual response. To compare the sex pheromone contents of atrial glands, pygidium, rectum, and colon, we recorded the EAG responses of male antennae to these extracts. Among the four tissues, colon extract caused the strongest EAG response. From gas chromatographic (GC) analysis, the quantities of periplanone-A (PA) and periplanone-B (PB) (the main sex pheromone components in female American cockroach) were 0.34 and 8.31 ng, respectively, in the colon of a virgin female. The ratio of PA to PB was ca. 1:24. The present results are different from those reported in a previous study, in which it was concluded that atrial glands are the sites of sex pheromone production in P. americana, and from GC analysis the quantity of PB was estimated to be 60 ng/female. There was no difference in sexual behaviors induced by colon and atrial glands of calling virgin females. It is unclear whether the colon is the sole sex pheromone source in the female American cockroach.     

Male sternal pheromone glands in Acanthosomatid shield bugs from Britain

Pheromone glands are present in the abdominal sternites of the pregenital region in the male adults in all British species of Acanthosomatidae. Secretory units are most numerous in the hawthorn bugAcanthosoma haemorrhoidale and maternal bugElasmucha grisea, next most numerous in the birch bugElasmostethus interstinctus, and least numerous in the juniper bugCyphostethus tristriatus. The largest dorsal abdominal scent glands were found inCyphostetheus. Ordinary dermal glands are also present. They are most numerous along the lateral margins of the abdomen. It is possible that Pendergrast's organ is the source of a female-produced pheromone. Secretory units were numerous in this organ inC. tristriatus female adults.     

Patterns of deposition of urine containing chemosignals that affect puberty and reproduction by wild stock male and female house mice (Mus domesticus)

House mice release chemosignals in their urine that influence the timing of puberty and reproductive condition in conspecific females. These experiments tested the possibility that mice depositing urine containing chemosignals that affect puberty and reproduction do so differentially with respect to urine cues from conspecifics. Mice were tested in cages containing samples of urine or water on cotton in wire mesh capsules protruding from the cage floor. Their urine deposition patterns were recorded on squares of filter paper positioned below the cages. Males deposited more urine than females housed in groups, estrous females, diestrous females, or prepubertal females. All groups of mice deposited urine in a nonrandom fashion with regard to urine cues from conspecifics. Male mice deposited more urine near locations previously soiled by females than near water or other males. Grouped females deposited more urine near male urine cues and avoided depositing urine near urine from other group-housed females. Both estrous and diestrous female mice deposited more urine near males than near other urine cues or water, possibly to attract mates. Prepubertal females avoided depositing their urine near male urine and urinated more near urine from grouped females than near other urine types or water. Young females may be avoiding possible male mates until they have attained puberty. This avoidance behavior may enhance the long-term reproductive success of the females that otherwise might mature and mate at too young an age.     

The sex pheromone glands ofDermacentor variabilis (Say) andDermacentor andersoni stiles

Evidence is presented indicating the presence of the tick sex pheromone, 2,6-dichlorophenol in lipid droplets in the foveal glands ofDermacentor variabilis (Say) andDermacentor andersoni Stiles. The pheromone appears to be dissolved in the lipid droplets. The droplets consist of neutral lipids, mostly triacylglycerides and cholesterol esters. The esterified fatty acid profiles of foveal gland triacylgylcerides are different from those of other tissues examined or reported for other ticks. The percentage of shorter chain, mostly saturated fatty acids were decreased, while the longer chain polyunsaturated fatty acids were increased. The biological advantages of lipid solution for storage, translocation, and release of the sex pheromone from the female tick are discussed.Supported by grant AI 10986, from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, U.S. Public Health Service, Department of Health, Education and Welfare, Bethesda, Maryland.     

A comparison of the effectiveness of the odors of rabbits,Oryctolagus cuniculus,in enhancing territorial confidence

In 14 series, each of 100 tests, involving 350 individual animals it has been shown that odors derived from the anal, inguinal and chin glands and urine strengthen to different degrees the confidence of European wild rabbits,Oryctolagus cuniculus. In the tests, two rabbits were put together on otherwise neutral ground previously treated with the odor from one of the contestants. The confidence of the rabbits was determined from an assessment of the record of the time to first physical contact, the frequencies of approaches and aggression, the number of attempts to escape, and also by making a subjective assessment of their general deportment. The results confirm earlier conclusions on the territorial functions of the anal and chin glands, the role of inguinal glands in individual identification, and the behavioral role of urine. On otherwise neutral ground the rabbits' own chin secretions were found to be the most effective in stimulating the confidence of males and that of their female partners. The odor of the anal gland secretions of males stimulated a similar but slightly weaker effect. The anal gland secretions of females influenced their own confidence but not that of males. Urine seemed to play a more important role in affecting the confidence of females than of males. Inguinal gland secretions had least effect. Odor from this source generally did not influence the confidence of the experimental animals; secretion from the inguinal glands of males had a slight effect on the behavior of their female partners.     

Misidentification by wild rabbits,Oryctolagus cuniculus,of group members carrying the odor of foreign inguinal gland secretion

The results of experiments designed to clarify the behavioral function of the odor of the secretion from the inguinal glands of rabbits,Oryctolagus cuniculus, are presented. Members of groups of female rabbits were smeared with odoriferous materials, and the behavior of their penmates towards them was recorded. Eight sources of natural rabbit odors and a commercial perfume were used in 180 tests involving 120 adult female wild-type rabbits. A high proportion of the animals smeared with the inguinal gland secretions from unfamiliar females were attacked by their pen-mates. In contrast, treatment with inguinal gland secretions from unfamiliar male rabbits; macerated inguinal gland materials from unfamiliar males and females, respectively; urine from unfamiliar females; and macerated anal gland material from unfamiliar females resulted in only minor disruption to the normal social relationships between the group members which was not significantly different to that caused by treatment with a commercial perfume.     

Some communicatory functions of scent marking in the cotton-top tamarin (Saguinus oedipus oedipus)

The ability of cotton-top tamarins to discriminate between scents from conspecifics and those from other tamarin species, and between scents from conspecific individuals was tested. Cotton-tops scent mark with specialized skin glands in the circumgenital area. Females possess larger glands than males and show more scent-marking behavior. In the first experiment, subjects were presented with a glass rod scented with either material collected from the surface of the scent glands of a conspecific female, with scent material from a female of a related species, or with an unscented rod. Glass rods carrying scent from conspecifics were sniffed more frequently than rods carrying scent from related tamarin species or unscented rods. A second experiment offered a choice between two glass rods, one scent marked by aSaguinus o. oedipus female, the second one scent marked by aSaguinus fusdcollis female. Shelves carrying rods that had been scent marked by conspecifics were contacted more frequently than those carrying rods marked by heterospecific females. Scent marks from conspecific females were also sniffed more frequently. A third experiment compared the level of responses to rods carrying material collected from the scent glands of female individuals to which the subjects had been habituated with their responses to rods carrying scent from females to which they had not been habituated. Contacting and sniffing responses to the scents of novel females were higher than those to the scents of females to which the subjects had been habituated.     

Analysis of gular and paracloacal gland secretions of the american alligator (Alligator mississippiensis) by thin-layer chromatography Gland,sex, and individual differences in lipid components

Secretions from the gular and the paracloacal glands of adult male and female American alligators (Alligator mississippiensis) were extracted with chloroform, weighed, and analyzed by thin-layer chromatography. In both sexes, more lipoidal secretions were recovered from the paracloacal glands than from the gular glands. Females produce more lipoidal secretions in both sets of glands than do males. The thin-layer chromatograms of extracts from both glands exhibit bands consistent with aliphatic alcohols, sterols, free fatty acids, and steryl esters. Triglycerides, hydrocarbons, phosphatidylethanolamine, and lysophosphatidylcholine also are indicated in some chromatograms. Gland, sex, and individual differences are suggested in the thinlayer profiles.     

Modulation of exploratory behavior in female mice by protein-borne male urinary molecules

Male pheromones are believed to attract females and repel male mice in open field tests but, when tested in more complex environments, they can attract male mice in usually avoided areas. Females were tested in an apparatus with one dark and one light side, in the absence or presence of male urine or the major urinary proteins (MUPs) bearing the natural ligands. Diestrous females were slower in leaving from the dark area when male urine or MUPs were present in it. Estrogen-primed females showed the opposite behavior, with an increase in the same latency. The light-avoidance behavior of prepubertal females, or females reared without males was not influenced by the presence of male chemosignals. The results show that adult female mice can react to MUP-borne volatiles as to adult male urine and use them as cues of male mice, if they were previously exposed to male cues during infancy. MUP-borne molecules are, thus, the olfactory trace of males in the environment and modulate mice exploratory behavior.     

Chemical characterization of urinary volatile compounds ofPeromyscus californicus,a monogamous biparental rodent

The urinary profiles of adult female and male California mice were examined to determine the volatile compounds that may affect reproduction. The urinary volatiles identified by gas chromatography-mass spectrometry (GC-MS) include ketones, pyrazines, alkanes, nitrile, and aldehyde. None of volatile compounds was specific to males or females. The concentration of urinary volatiles in pregnant and lactating females was significantly higher than in virgin females. Male urinary volatile concentrations were similar to those of pregnant and lactating females. The GC profiles of both sexes were distinguished by a high percentage (36%) of pyrazine derivatives that were also in unusually high concentrations. These compounds may play a wide-ranging role in the control of reproduction in the California mouse.     

Evidence for a compound in Comstock-Kellog glands modulating premating behavior in male desert locust,Schistocerca gregaria

The Comstock-Kellog glands in adult females of certain acridid species, including the desert locust, Schistocerca gregaria have been implicated as a source of volatiles that play a role in mating behavior. A dichloromethane extract of the glands was analyzed for metabolites by gas chromatography, coupled gas chromatography–electroantennographic detection, and mass spectrometry. Coupled gas chromatography–electroantennographic detector (GC-EAD) analysis revealed a component that elicited an electroantennogram response from the antenna of adult male S. gregaria. The compound was identified by GC-MS as pentanoic acid. The levels of the compound in the gland extract varied with age of female locust; it was present in detectable amounts only in 14- to 16-day-old females. In bioassays, pentanoic acid significantly stimulated pre-mating behavior in male desert locust. These results are discussed in relation to the biology of the locust.     

Increased scent marking in male Mongolian gerbils by urinary polypeptides of female conspecifics

Male Mongolian gerbils scent mark during a standardized openfield test on an individually different but more-or-less constant level in the absence of females. The presence of females in the same room but without direct contact to the males increases the marking frequencies of the males to an individually different higher level. A similar increase in marking frequency is obtained by application of female urine directly to the nostrils of the males, but not by male gerbil urine or female laboratory mouse urine. This indicates that substances present in female gerbil urine are responsible for the increase in male marking behavior. Using this bioassay, various fractions of female urine were tested for their influence on marking behavior. Our results to date show that the component increasing male scent-marking behavior is hydrophilic and nonvolatile and is contained in the nondialyzable, high-molecular-weight polypeptide fraction of female gerbil urine.     

Castor sacs and anal glands of the north american beaver (Castor canadensis): their histology,development, and relationship to scent communication

Both sexes of beavers possess a pair of castor sacs and a pair of anal glands located in paired subcutaneous cavities between the pelvis and the base of the tail. The castor sacs are not glandular in the histological sense, hence references to these structures as preputial glands or castor glands are misnomers. The wall of the castor sacs is plicate and comprised of three distinct zones: an outer layer of vascular connective tissue, a two-to five-cell-thick layer of mitotic epithelial cells, and several densely packed layers of cornified epithelium which grade into more widely separated sheets toward the lumen. Monocultures of a gram-positive facultatively anaerobic bacterium were present in the lumen of all castor sac preparations. Differences in the frequency of castoreum deposition were not attributable to differences in the structure of the castor sacs. The anal glands of beavers are holocrine sebaceous glands. These glands develop more rapidly than the castor sacs. Anal gland tissue from embryos exhibited cellular characteristics associated with the production of sebum. Secretory activity was evident in all preparations. The relationship of castoreum and anal gland secretion to scent communication among beavers is discussed.     

Scent marks as social signals inGalago crassicaudatus I. Sex and reproductive status as factors in signals and responses

In simultaneous presentations of the scent marks of male and female conspecifics, thick-tailed galagos (Galago crassicaudatus) of both sexes responded most strongly to the female scent. Males differentiated between the scents most clearly in their sniffing, females in their scent marking in response. The scent of estrus females elicited increased licking of the scent by both sexes. This greater response to female scent and the contrasts in response patterns between the sexes are discussed and related to galago social and spatial relationships in the wild.     

On-line Thermal Desorption–Gas Chromatography of Intact Insects for Pheromone Analysis

By using a thermodesorption system (TDS) together with a programmable temperature vaporizer (PTV) injector, we confirmed the composition of the sex pheromone of Adoxophyes orana (Lepidoptera: Tortricidae) and Campylomma verbasci (Heteroptera: Miridae) from a single insect per analysis. Intact females and males or pheromone glands were placed in the oven part of the TDS, which was subsequently heated. The compounds released were transferred to the PTV, which was cooled to –150°C. Injection was on a dual-column GC by heating the PTV rapidly to 250°C. The major sex pheromone compounds of A. orana were found only in the pheromone gland of females. Male and female C. verbasci showed fingerprint-identical chromatograms, except for the two sex pheromone compounds, which were present only in females. No distinct differences were found in compounds released from female and male Lygocoris pabulinus (Heteroptera: Miridae). The advantages of this rapid method are the high sensitivity and the low degree of degradation and contamination. This technique was effective in analyzing small insects by gas chromatography–mass spectrometry (GC-MS) without prior manipulation, such as solvent extraction or distillation.