Ste2p and Ste3p are well-characterized yeast pheromone G-protein Coupled Receptors (GPCR) those are involved in the signaling of mating responses that lead to cell fusion. Their signaling–associated interactions with G-protein/MAPK signal transduction machinery are well established, homologous to those in mammalian systems, and serve as a simplified model system in GPCR research. While the arrestin- mediated biased signaling mechanism of mammalian GPCR has not been discovered for the pheromone receptors, a recent demonstration of α-arrestins being involved in the internalization of the pheromone GPCR, Ste2p was reported. The present study was designed to reevaluate and extend the alternate functionality for pheromone receptors and to determine the role of yeast arrestins in the yeast mating. Specific residues in the TM6 of Ste2p exhibiting strong mating and constitutive MAPK signaling were combined and investigated in terms of their effect on MAPK signal transduction leading to cell cycle arrest as well as their impact on downstream mating projection formation and zygote formation events. Our findings indicate that Ste2p possess as specific residues that govern its relative bias for mediating MAPK signaling or mating events. Relative dose response experiments accounting for systemic and observation bias for these mutations yielded evidence of mutational-derived functional biases for Ste2p and further validated the alternate pheromone dependent functionalities for Ste2p. Further, arrestin knockout and knock-in studies showed that Art1 (Ldb19) is selectively involved in the regulation of zygote formation but not MAPK signal transduction following the binding of ligand to Ste2p receptors. In addition, ligand stimulated selective localization of Art1 (Ldb19) to the mating projection, implicating it in the regulation of downstream mating functionalities. Overall, while leaving the full mechanism of alternate/biased Ste2p signaling to be elucidated, these results highlight the possibility of continued relevance of the yeast pheromone-mating pathway as a simplified model for GPCR research in the context of arrestin-mediated biased GPCR signaling.

In this master thesis we investigate, from a topological point of view and without applying Thurston´s Theorem, why the mating of the so called basilica polynomial <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D(z)=z%5E%7B2%7D-1" /> and the dendrite <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7Bi%7D(z)=z%5E%7B2%7D+i" /> is shared with the mating of <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D" /> and the dendrite <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-i%7D(z)=z%5E%7B2%7D-i" />. Both these matings equal the rational map <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?R_%7B3%7D(z)=%5Cfrac%7B3%7D%7Bz%5E%7B2%7D+2z%7D" />. Defined in the thesis are for both matings homeomorphic changes of coordinates<img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?%5Cpsi_%7B-1%7D%5E%7B%5Cpm%7D" /> from the set <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?L=%5Coverset%7B%5Ccirc%7D%7BK%7D%5Cleft(f_%7B-1%7D%20%5Cright)%5Ccup%5Cleft(%5Ccup_%7Bn=0%7D%5E%7B%5Cinfty%7Df_%7B-1%7D%5E%7B%5Ccirc(-n)%7D(z_%7B%5Calpha%7D)%5Cright)" /> to the Fatou and Julia set of <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?R_%7B3%7D" />. Here <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?K%5Cleft(f_%7B-1%7D%20%5Cright)" /> is the filled Julia set of <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D" /> and <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?z_%7B%5Calpha%7D" /> is the <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?%5Calpha" />-fixed point of <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?K%5Cleft(f_%7B-1%7D%20%5Cright)" />. / I detta examensarbete undersöker vi, från en topologisk synvinkel och utan applicering av Thurstons teorem, varför matchningen av det så kallade basilikapolynomet <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D(z)=z%5E%7B2%7D-1" /> och dendriten <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7Bi%7D(z)=z%5E%7B2%7D+i" /> är delad med matchningen av <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D" /> och dendriten <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-i%7D(z)=z%5E%7B2%7D-i" />. Båda dessa matchningar är lika med den rationella avbildningen  <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?R_%7B3%7D(z)=%5Cfrac%7B3%7D%7Bz%5E%7B2%7D+2z%7D" />. Definierat i examensarbetet är för båda matchningarna homoemorfa koordinatbyten<img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?%5Cpsi_%7B-1%7D%5E%7B%5Cpm%7D" /> från mängden<img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?L=%5Coverset%7B%5Ccirc%7D%7BK%7D%5Cleft(f_%7B-1%7D%20%5Cright)%5Ccup%5Cleft(%5Ccup_%7Bn=0%7D%5E%7B%5Cinfty%7Df_%7B-1%7D%5E%7B%5Ccirc(-n)%7D(z_%7B%5Calpha%7D)%5Cright)" /> till Fatou- och Juliamängden av <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?R_%7B3%7D" />. Här är <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?K%5Cleft(f_%7B-1%7D%20%5Cright)" /> den ifyllda Juliamängden av avbildningen <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?f_%7B-1%7D" /> och <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?z_%7B%5Calpha%7D" /> är den <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?%5Calpha" />-fixerade punkten i <img src="http://www.diva-portal.org/cgi-bin/mimetex.cgi?K%5Cleft(f_%7B-1%7D%20%5Cright)" />.

Mate choice among males is relatively understudied, despite recent evidence supporting its ubiquity. Theory predicts male mate choice in response to variation in female quality, and male mating strategies that limit polygyny. However empirical research investigating these connections, particularly under natural conditions, is generally lacking. Using the Western black widow spider, Latrodectus hesperus, I investigated male mate choice, male investment, and the advantage of mating with females that differ in their potential to deliver fitness benefits to males via higher fecundity and/or reduced risk of sperm competition. Males were found to be preferentially attracted to larger, high-diet virgin females over females that were smaller due to a lower diet, or having been previously mated, or both. Through a three-year field study on females I found that males likely benefit from selectivity, as smaller, low-diet females often failed to deposit any egg sacs throughout the breeding season, likely due to their shortened web-site tenure and/or lack of bodily resources. I investigated the costs and benefits to male genital breakage in L. hesperus, a strategy typically assumed to result in male sterility while providing paternity protection. I found that genital mutilation in L. hesperus did not cause sterility and that males were capable of inseminating multiple females, likely because of the comparatively low amount of genital damage and the likelihood of the efficient placement of genital fragments. Evidence from double mating trials supported the efficacy of broken genital fragments as plugs, but first male sperm precedence was often maintained in cases where male genital breakage failed or fragments were positioned incorrectly, which occurred frequently. Together, these laboratory and field experiments contribute to a more complete view of mate choice.

Mate choice among males is relatively understudied, despite recent evidence supporting its ubiquity. Theory predicts male mate choice in response to variation in female quality, and male mating strategies that limit polygyny. However empirical research investigating these connections, particularly under natural conditions, is generally lacking. Using the Western black widow spider, Latrodectus hesperus, I investigated male mate choice, male investment, and the advantage of mating with females that differ in their potential to deliver fitness benefits to males via higher fecundity and/or reduced risk of sperm competition. Males were found to be preferentially attracted to larger, high-diet virgin females over females that were smaller due to a lower diet, or having been previously mated, or both. Through a three-year field study on females I found that males likely benefit from selectivity, as smaller, low-diet females often failed to deposit any egg sacs throughout the breeding season, likely due to their shortened web-site tenure and/or lack of bodily resources. I investigated the costs and benefits to male genital breakage in L. hesperus, a strategy typically assumed to result in male sterility while providing paternity protection. I found that genital mutilation in L. hesperus did not cause sterility and that males were capable of inseminating multiple females, likely because of the comparatively low amount of genital damage and the likelihood of the efficient placement of genital fragments. Evidence from double mating trials supported the efficacy of broken genital fragments as plugs, but first male sperm precedence was often maintained in cases where male genital breakage failed or fragments were positioned incorrectly, which occurred frequently. Together, these laboratory and field experiments contribute to a more complete view of mate choice.

No description available.

Many Australian marsupial species require active population management to ensure their survival in the wild. Such management should be based on a sound understanding of species biology. This thesis explores how knowledge of reproduction and genetics can be applied to the management of three Australian marsupial species faced with contrasting management scenarios. The ??vulnerable?? greater bilby is the sole remaining desert bandicoot in Australia. They are a secretive, solitary species whose mating system is unclear. This research examined temporal changes in genetic diversity within two captive breeding programs utilising different management strategies. Using seven microsatellite loci, this study found the regular translocation of new individuals into the population maintained genetic diversity. Parentage analysis revealed the bilby to have a promiscious mating system. Sires and non-sires could not be distinguished by morphological traits. The tammar wallaby is a polygynous, solitary species that is threatened on mainland Australia, but overabundant on some offshore islands. The population genetics of tammars from the Abrolhos Islands in Western Australia were examined using nine autosomal and four Y-linked microsatellite loci, and mitochondrial DNA. There was a relationship between island size, population size and genetic diversity. The Abrolhos populations have significantly lower genetic diversity and are more inbred than mainland tammars and all sampled populations were significantly differentiated. The Abrolhos and mainland populations should be treated as separate Management Units. The eastern grey kangaroo is a gregarious, polygynous species that is often locally overabundant. To determine traits influencing male reproductive success, behavioural, morphological, physiological and genetic data were examined and showed dominance status, body size and testosterone concentrations were important factors. Sires were also significantly more heterozygous and genetically dissimilar to females, than non-sires. As body condition influences individual fitness, and management decisions; five body condition indices (BCI) calculated from morphological data were validated using serum biochemistry and haematology in two kangaroo populations with contrasting body condition. Blood parameters were found to be more reliable indicators of condition, questioning the credibility of BCIs currently used in management. These studies demonstrate the importance of reproductive and genetic data in assisting wildlife management, regardless of a species conservation status.

This study investigates the reproductive strategies of the pill-box crab, Halicarcinus cookii on the Kaikoura Peninsula, New Zealand. Various aspects essential to understanding reproductive strategies were examined including growth, population dynamics, reproductive biology and mating behaviour. H. cookii exhibits obvious sexual dimorphism such that females develop wide abdomens forming brood chambers, and males tend to grow larger than females and have larger chelipeds in relation to body size. H. cookii allocates energy into growth and reproduction in separate phases of its life cycle where growth ceases as reproductive maturity begins due to a terminal/pubertal moult. Despite the presence of ovigerous females throughout the 15 month sampling period, the population was highly seasonal, with peaks in recruitment and growth occurring primarily during the winter months and peaks in numbers of mature individuals during the summer months. Reproductive output increased with body size in H. cookii, as larger females produced more eggs and larger males transferred more sperm than their smaller counterparts. Ovaries matured prior to the terminal/pubertal moult (anecdysis) and, in multiparous females, in synchrony with brood development, allowing females to produce broods in quick succession, maximising their reproductive output in their short life span (approximately 12-18 months, 6 months as an adult). Incubation duration of broods decreased as seawater temperature increased, suggesting that temperature is the primary cause of the seasonal population cycling. Sperm storage allowed females to produce at least 4 fertilised broods without re-mating. Some sperm mixing in the spermathecae appeared to occur and the ventral-type structure implies last male sperm precedence. Males therefore preferentially mated with females closest to laying a new brood and guarded them longer than other females to ensure their paternity. Guarding duration varied according to the sex ratio allowing males to maximise their reproductive output.

The reproductive strategies of four intertidal grapsid crabs, Hemigrapsus sexdentatus, H. crenulatus, Cyclograpsus lavauxi, and Helice cressa, were studied in the field and laboratory, with emphasis on mating behaviour, duration of female receptivity, and sperm competition. Mating occurred in all species during the intermoult on the days prior to oviposition, when the gonoporo opercula of females became temporarily mobile. Female Helice crassa mated up to three weeks after oviposition, but in all other species mating typically ceased at egg-laying. Male Hemigrapsus pp. used a female-centered competition strategy in which they searched for and defended receptive females until they laid eggs. In contrast, male C. lavauxi searched for and intercepted receptive females only for the duration of copulation and then pursued other receptive females (a mating system termed encounter rate competition with pure search and interception). Male Helice crassa searched for receptive females in their immediate neighbourhood and mated with them briefly on the substrate or in the burrow after which the female left (a mating system termed encounter rate competition with neighbourhoods of dominance). The mating season was short and highly synchronous for Hemigrapsus exdentatus and Cyclograpsus lavauxi and asynchronous for Hemigrapsus crenulatus and Helice crassa. In the laboratory, the mean duration of receptivity for females housed with three males varied between 4.1 and 12.4 days, and the copulation frequency of females varied before oviposition between 2.1 and 24.3 times (mean) depending on the species. Female Hemigrapsus spp. isolated from males stayed receptive significantly longer than females held continuously with males. This suggests that females are able to control the duration of their receptivity, and therefore the time available for mating, according to the absence or presence of males. The operational sex ratio (OSR) had no effect on the duration of female receptivity, but female Hemigrapsus crenulatus mated more often when several males were competing for access. Therefore, male-male competition increased the number of matings per female and hence sperm competition within the female spermathecae. Larger males mated significantly more often than smaller males in all species. However, male size did not affect ejaculate size, meaning that small and large males transferred similar-sized ejaculates, e.g., in Hemigrapsus spp. Males of the two Hemigrapsus species followed a different strategy of sperm allocation. Male H. crenulatus, which are typically confronted with a high mating frequency of the female and a long, asynchronous mating season, distributed similar-sized ejaculates, irrespective of female size. By contrast, male H. sexdentatus, which experience a comparatively lower risk of sperm competition during a short, synchronised mating season, invested larger ejaculates for larger females than for smaller females. In addition, the size of the first and second ejaculates transferred to a female by a male H. crenulatus were not significantly different, whereas the first was larger than the second for H. sexdentatus. A parasitological survey was undertaken of the four grapsid crabs and the presence, seasonal variation and relationship with host gender and size of parasites determined. Four internal parasites were discovered: Nectonema zealandica n. Sp. (Nematomorpha: Nectonematoidea), portunion sp. (Isopoda: Entoniscidae), Profilicollis novaezelandensis n. sp. and profilicollis antarcticus (Acanthocephala: P olymorphidae). Portunion sp. castrated its female hosts, but not the males thereby creating a more male-biased sex ratio. Males parasitised with portunion sp. were equally successful during male-male competition and the number of matings they achieved. The above findings are important for our current understanding of mating strategies in Grapsidae, which are more diverse than previously thought. Females with a restricted duration of sexual receptivity have some control over their receptive period and can therefore influence the OSR and the extent of male-male competition. As females mated multiple times during their receptive period, sperm competition is a common feature in Grapsidae. However, males employed different tactics in regards to sperm competition such as longer mating duration (e.g., C. lavauxi), high number of matings (Helice crassa), or post-copulatory mate guarding until oviposition (Hemigrapsus spp.).

The honey possum Tarsipes rostratus, a marsupial endemic to South-Western Australia, feeds exclusively upon nectar and pollen. It is one of the smallest marsupials, with adult females (8-12g) significantly larger than adult males (6-9g). Honey possum males have the longest sperm (356µm) recorded for any mammaland the testes represent 4.2% body weight, amongst the largest recorded formammal species. These features suggest that sperm competition is an importantpart of the mating system. This study used a combination of field based studies,DNA analysis and histological examination of the female reproductive tract toinvestigate the life history, multiple paternity and reproduction of the honey possum innatural populations in the Fitzgerald River National Park (FRNP), on the south coastof Western Australia. This study drew upon earlier work on the honey possum in the FRNP in order to describe its life-history. The honey possum is short-lived (1-2 years), and attains sexual maturity whilst still growing. All four teats are occupied after birth, but the litter is reduced to 2 or 3 young during pouch life. The young have a relatively slow rate of growth. Breeding occurs continuously throughout the year, but is affected by the flowering phenologies of its foodplants. The greatest proportion of females with pouch-young occurs in winter; there are fewest pouch-young in autumn, a time of year when there is a dearth of flowers. Honey possums are essentially solitary animals, with no structured social unit, and male and female home ranges overlap. In captivity they are largely tolerant of one another, but larger females are behaviorally dominant to smaller females and to males. The densities and structure of the honey possum populations in the FRNP were analyzed from trapping data collected over 19 years. Population densities fluctuated significantly from season to season throughout the year, with changes in the flowering food resources available. There were also year-to-year differences in the intensity of those fluctuations, and these were significantly associated with rainfall in the previous year, and probably mediated through a lag effect in the flowering of the honey possum’s foodplants. The greatest densities of animals occurred over winter. In years following high rainfall, mean winter densities reached 88 individuals per hectare. The lowest densities occurred in spring, and in years following low rainfall mean spring densities fell to 8 individuals per hectare. Even at these lowest densities, there is still the potential for interaction between males and females. A succession from high to low, then back to high densities was seen during the three years of the present study (2000-2002) and this shadowed a similar succession of changes in rainfall. The proportion of females with pouch-young was significantly affected by the season, and by rainfall in the previous year. Years following low rainfall had a lower proportion of females in a condition to breed. The autumn dip in breeding that occurred in all years was exacerbated following dry years. Of those females that did breed in 2001, a time of low resources, there was no difference in the size of the litter compared to 2000 and 2002, times of higher resource availability. The sex-ratio of pouch young was at parity, but there was a slight bias towards males among both juveniles (56%) and adults (58%). This was probably due to the greater movements shown by males. Sex ratios were not affected by changes in rainfall and density. Male-biased dispersal was detected using genetic data and the movement patterns of males showed that they moved greater distances than females during their normal activity. Analysis of four microsatellite loci revealed extremely high levels of variation, with 28 to 50 alleles per locus and a mean expected heterozygosity of 0.95. These are amongst the highest seen in any microsatellite study of vertebrates. There was multiple paternity in 86% litters, using a minimum number of sires per litter method, and in 95% litters, using an estimated number of sires method based upon the relatedness of litter males. This indicates that multiple mating is frequent in female honey possums and is evidence for sperm competition. The estimated number of sires in a litter was often three or four. In 41% of cases, the number of sires was less than the number of young in the litter, indicating that some males were more successful at siring offspring than others. Nevertheless, no more than two offspring in a litter were known to have been sired by the same male. Despite marked fluctuations in density from high in 2000, to low in 2001, then high again in 2002, the level of multiple paternity remained equally high in all years. Embryonic diapause and female reproduction was investigated in the honey possum. All adult females examined, both with and without pouch-young, were either close to oestrus, had ovulated or were carrying conceptuses. The honey possum has a postpartum oestrus and it was evident that this occurs approximately 2-4 days after birth. Cleavage and formation of the unilaminar blastocyst appears to occur rapidly over approximately 5 days. Embryonic diapause proceeded in a two phase manner similar to other small possum species. The unilaminar blastocyst expanded rapidly at first; and then, from about 18-20 days after birth, the diameter of the blastocyst remained constant at approximately 1.2-1.8mm. No growth or development beyond the unilaminar stage was observed during pouch-life. The first signs of reactivation occurred during lactation, after pouch exit, and expansion of the blastocyst only occurred in one post-lactational female. The development of the corpus luteum appeared different to patterns described for other marsupials, but its formation coincided with the formation of the unilaminar blastocyst. The diameter of the corpus luteum remained constant throughout diapause. The histology of the reproductive tract was generally similar to other marsupials. There were no sperm storage crypts in the female reproductive tract. The length of pouch-life in the honey possum was 55-65 days, and the interval between litters of the same size varied between 65 and 100 days. Embryonic diapause may reduce the time between production of successive litters in the honey possum, but lifetime reproductive potential is reasonably low. Females had up to four litters over the period that they were captured. Thus, each litter represents a substantial proportion (25%) of a female’s lifetime reproductive output. Reproductive amortization occurred, with 61% loss overall, due to overproduction of ova, loss of conceptuses and reduction of the litter during lactation. The behavioural dominance of females suggests that multiple mating is an active strategy, and this presumably allows the genetic quality of their offspring to be maximized. Males that succeed in sperm competition may be of better intrinsic quality. Overproduction of conceptuses by females presents the opportunity for them to select those fertilized by intrinsically viable males or genetically compatible males. Sexually active males are present all year round. Females were not synchronous in their sexual receptivity, and this would lead to a skewed operational sex ratio, with more reproductive males than oestrous females. Since adult males are significantly smaller than adult females and possess no ornaments or armaments, it is unlikely that males overtly fight for access to females. Rather, males appear to monitor the reproductive status of females through smell, and probably compete in their ability to locate oestrous females. The risk and intensity of sperm competition is high, sexual selection for a large investment in spermatogenesis is evident and competition after copulation is probably an important factor in the mating system. It is likely that males, as well as females mate multiply, and the mating system is promiscuous.

Many Australian marsupial species require active population management to ensure their survival in the wild. Such management should be based on a sound understanding of species biology. This thesis explores how knowledge of reproduction and genetics can be applied to the management of three Australian marsupial species faced with contrasting management scenarios. The ??vulnerable?? greater bilby is the sole remaining desert bandicoot in Australia. They are a secretive, solitary species whose mating system is unclear. This research examined temporal changes in genetic diversity within two captive breeding programs utilising different management strategies. Using seven microsatellite loci, this study found the regular translocation of new individuals into the population maintained genetic diversity. Parentage analysis revealed the bilby to have a promiscious mating system. Sires and non-sires could not be distinguished by morphological traits. The tammar wallaby is a polygynous, solitary species that is threatened on mainland Australia, but overabundant on some offshore islands. The population genetics of tammars from the Abrolhos Islands in Western Australia were examined using nine autosomal and four Y-linked microsatellite loci, and mitochondrial DNA. There was a relationship between island size, population size and genetic diversity. The Abrolhos populations have significantly lower genetic diversity and are more inbred than mainland tammars and all sampled populations were significantly differentiated. The Abrolhos and mainland populations should be treated as separate Management Units. The eastern grey kangaroo is a gregarious, polygynous species that is often locally overabundant. To determine traits influencing male reproductive success, behavioural, morphological, physiological and genetic data were examined and showed dominance status, body size and testosterone concentrations were important factors. Sires were also significantly more heterozygous and genetically dissimilar to females, than non-sires. As body condition influences individual fitness, and management decisions; five body condition indices (BCI) calculated from morphological data were validated using serum biochemistry and haematology in two kangaroo populations with contrasting body condition. Blood parameters were found to be more reliable indicators of condition, questioning the credibility of BCIs currently used in management. These studies demonstrate the importance of reproductive and genetic data in assisting wildlife management, regardless of a species conservation status.