Plasma Esterase (ES) Polymorphism in the Tammar Wallaby, Macropus eugenii

The major plasma esterase in the tammar wallabywas identified as a carboxylesterase by inhibitionstudies and polymorphism with six variants was observedby isoelectric focusing (pH 4.2-4.9), followed by staining for esterase activity. Familystudies demonstrated an inheritance of six codominantalleles, ES^A,B,C,D,E,F, and populationstudies revealed marked differences in the allelefrequencies in five Australian populations of tammar wallabies.     

Milk Composition during Lactation Suggests a Mechanism for Male Biased Allocation of Maternal Resources in the Tammar Wallaby (Macropus eugenii)

Recent research has found empirical evidence in support of the Trivers-Willard Hypothesis that offspring sex allocation is correlated with maternal investment. Tammar wallabies birthing sons have higher investment ability; however a mechanism for sex specific differential allocation of maternal resources in wallabies remains elusive. In metatherians the majority of maternal investment is during lactation. To examine if differential allocation occurs during lactation, we measured total milk protein, lipid and carbohydrates, from mothers with male and female pouch young, during phase 2B (100–215 days post partum) and phase 3 (215–360 days post partum) of lactation. Mothers of sons allocated significantly higher levels of protein than mothers of daughters during phase 2B of lactation, however no sex specific difference in maternal allocation was found for lipids, carbohydrates, or any milk component during phase 3 of lactation. We were unable to measure milk production to establish any differences in the amount of milk allocated. However, with the production of more milk comes a dilution effect on milk components. Given that we find no apparent dilution of milk components may suggest equality in milk production. Offspring body weight at 14 months of age was related to protein allocation during phase 2B of lactation, providing a maternal mechanism for differential allocation with fitness consequences. We believe collection of earlier phase 2A (0–100 days post partum) milk may yield important results given that differential investment in metatherians may be most apparent early in lactation, prior to any significant maternal investment, when a decision on termination of investment can be made with very little energetic loss to the mother. Interestingly, small mothers did not birth sons and better maternal condition was associated with raising sons. These data are in support of TWH and demonstrate a potential mechanism through which condition dependent and sex specific maternal investment may occur.     

From Embryo to Adult: The Complete Development and Unusual Replacement of the Dentition of the Tammar Wallaby (Macropus eugenii)

Unlike their reptile-like ancestors with continuous tooth replacement, mammals have evolved to replace each tooth either only once, or not at all. In previous large-scale comparative studies, it has been suggested that this tooth replacement only occurs from a successional dental lamina produced lingually to the primary tooth. This study aims to document the complete tooth development and replacement pattern of the tammar wallaby (Macropus eugenii). The tammar wallaby is a diprotodont marsupial, a group defined by their two procumbent lower incisors. To provide a comprehensive documentation of the spatio-temporal pattern of tooth development, we used Lugol’s Iodine staining and microCT scanning (diceCT) of embryos and pouch young into adulthood, resulting in high resolution 3D models for both soft and mineralised stages of development for all tooth positions. Our results reveal that the eponymous lower incisors are the successional generation at the third incisor locus, where the primary dentition initiates but never erupts. Furthermore, we track the development of the only replacement tooth, the permanent third premolar (P3), from initiation to eruption, and found it develops from the primary dental lamina, mesial to the dP3. This is contrary to the conventional view of lingual replacement from successional lamina in mammals. Our findings indicate that no functional tooth replacement occurs in the tammar wallaby, and expands the diversity of tooth replacement patterns found in mammals. We also conclude that since almost all marsupial and placental mammals produce replacement teeth from the distalmost deciduous premolar, this tooth should be considered homologous in these two groups.

     

The marsupial MHC: The Tammar Wallaby,Macropus eugenii,contains an expressed DNA-like gene on chromosome 1

Isolates of cauliflower mosaic virus (CaMV) differ in host range and symptomatology. Knowledge of their sequence relationships should assist in identifying nucleotide sequences responsible for isolate-specific characters. Complete nucleotide sequences of the DNAs of eight isolates of CaMV were aligned and the aligned sequences were used to analyze phylogenetic relationships by maximum likelihood, bootstrapped parsimony, and distance methods. Isolates found in North America clustered separately from those isolated from other parts of the world. Additional isolates, for which partial sequences were available, were incorporated into phylogenetic analysis of the sequences of genome segments corresponding to individual protein coding regions or the large intergenic region of CaMV DNA. The analysis revealed several instances where the position of an isolate on a tree for one coding region did not agree with the position of the isolate on the tree for the complete genome or with its position on trees for other coding regions. Examination of the distribution of shared residue types of phylogenetically informative positions in anomalous regions suggested that most of the anomalies were due to recombination events during the evolution of the isolates. Application of an algorithm that searches for segments of significant length that are identical between pairs of isolates or contain a significantly high concentration of polymorphisms suggested two additional recombination events between progenitors of the isolates studied and an event between the XinJing isolate and a CaMV not represented in the data set. An earlier phylogenetic origin for CaMV than for carnation etched ring virus, the caulimovirus used as outgroup in these analyses, was deduced from the position of the outgroup with North American isolates in some trees, but with non-North American isolates in other trees. Correspondence to: U. Melcher     

An immunomodulator used to protect young in the pouch of the Tammar wallaby, Macropus eugenii

Eugenin [pGluGlnAspTyr(SO(3))ValPheMetHisProPhe-NH(2)] has been isolated from the pouches of female Tammar wallabies (Macropus eugenii) carrying young in the early lactation period. The sequence of eugenin has been determined using a combination of positive and negative ion electrospray mass spectrometry. This compound bears some structural resemblance to the mammalian neuropeptide cholecystokinin 8 [AspTyr(SO(3))MetGlyTrpMetAspPhe-NH(2)] and to the amphibian caerulein peptides [caerulein: pGluGlnAspTyr(SO(3))ThrGlyTrpMetAspPhe-NH(2)]. Eugenin has been synthesized by a route which causes only minor hydrolysis of the sulfate group when the peptide is removed from the resin support. Biological activity tests with eugenin indicate that it contracts smooth muscle at a concentration of 10(-9) M, and enhances the proliferation of splenocytes at 10(-7) M, probably via activation of CCK(2) receptors. The activity of eugenin on splenocytes suggests that it is an immunomodulator peptide which plays a role in the protection of pouch young.     

Pinning down a polymorphic parasite: new genetic and morphological descriptions of Eimeria macropodis from the Tammar wallaby (Macropus eugenii)

Identification of the protozoan parasite, Eimeria has traditionally relied on oocyst morphology, host range and life-cycle attributes. However, it is increasingly recognized that Eimeria species can vary in size and shape across their host range, an attribute known as 'polymorphism' that presents a unique challenge for identification. Advances in molecular tools hold promise for characterising Eimeria that may otherwise be misclassified based on morphology. Our study used morphologic and molecular traits of the oocyst life stage to identify a polymorphic parasite, Eimeria macropodis in a captive Tammar wallaby (Macropus eugenii) population in Australia. Molecular characterization highlighted the need to use multiple genetic markers (18S SSU and cytochrome c oxidase subunit I) to accurately identify E. macropodis owing to heterozygous alleles at the 18S SSU locus. This study provided an opportunity to assess the utility and shortcomings of morphologic and molecular techniques for 'pinning down' a polymorphic species. Moreover, our study was able to place E. macropodis in an evolutionary context and enhance resolution of the under-studied marsupial clade.     

Component D of chicken hemoglobin and the hemoglobin of the embryonic Tammar wallaby (Macropus eugenii) self-associate upon deoxygenation: Effect on oxygen binding

Extensive measurements of oxygen binding by some vertebrate hemoglobins (Hbs) have suggested an unusually high degree of cooperativity with reported Hill coefficients, n(H), greater than 4.0. We have reexamined this possibility of "super-cooperativity" with chicken Hb components A (alpha(A) (2)beta(2)) and D (alpha(D) (2)beta(2)). Prior studies have shown that component D but not A self-associates to dimers of tetramers upon deoxygenation. This self-association is reflected in the oxygen equilibrium of Hb D which shows a maximal n(H), greater than 4.0 at approximately 4 mM heme concentration. In contrast, component A has maximal n(H) value below 3. The value of the maximal n(H) for Hb D increases linearly with the fraction of octamer present in the deoxy Hb. We anticipate that deoxygenation-dependent self-association will be shown to be a general property of Hb D from birds and reptiles. Neither oxygen equilibria nor sedimentation measurements show any evidence that components A and D interact to form a complex when deoxygenated. We have also reexamined the oxygen equilibria of Hbs of an embryonic marsupial, the wallaby. The equilibria in red cells have been reported to have Hill coefficients as high as 5-6. Although our oxygen equilibrium measurements of solutions of unfractionated wallaby Hb at a concentration of approximately 1 mM show no n(H) values greater than approximately 3.0, sedimentation velocity measurements provide clear evidence for deoxygenation-dependent self-association.     

Control of jaw movements in two species of macropodines (Macropus eugenii and Macropus rufus)

The masticatory motor patterns of three tammar wallabies and two red kangaroos were determined by analyzing the pattern of electromyographic (EMG) activity of the jaw adductors and correlating it with lower jaw movements, as recorded by digital video and videoradiography. Transverse jaw movements were limited by the width of the upper incisal arcade. Molars engaged in food breakdown during two distinct occlusal phases characterized by abrupt changes in the direction of working-side hemimandible movement. Separate orthal (Phase I) and transverse (Phase II) trajectories were observed. The working-side lower jaw initially was drawn laterally by the balancing-side medial pterygoid and then orthally by overlapping activity in the balancing- and working-side temporalis and the balancing-side superficial masseter and medial pterygoid. Transverse movement occurred principally via the working-side medial pterygoid and superficial masseter. This pattern contrasted to that of placental herbivores, which are known to break down food when they move the working-side lower jaw transversely along a relatively longer linear path without changing direction during the power stroke. The placental trajectory results from overlapping activity in the working- and balancing-side adductor muscles, suggesting that macropods and placental herbivores have modified the primitive masticatory motor pattern in different ways.     

Cranial osteogenesis in Monodelphis domestica (Didelphidae) and Macropus eugenii (Macropodidae)

The pattern of onset and general rate of cranial ossification are compared in two marsupials, Monodelphis domestica (Didelphidae) and Macropus eugenii (Macropodidae). In both species a similar suite of bones is present at birth, specifically those surrounding the oral cavity and the exoccipital, and in both postnatal events follow a similar course. The facial skeleton matures more rapidly than the neurocranium, which is characterized by an extended period of ossification. Most dermal bones begin ossification before most endochondral bones. Endochondral bones of the neurocranium are particularly extended in both the period of onset of ossification and the rate of ossification. These data confirm suggestions that morphology at birth is conservative in marsupials and we hypothesize that the pattern of cranial osteogenesis is related to two distinct demands. Bones that are accelerated in marsupials are correlated with a number of functional adaptations including head movements during migration, attachment to the teat, and suckling. However, the very slow osteogenesis of the neurocranium is probably correlated with the very extended period of neurogenesis. Marsupials appear to be derived relative to both monotreme and placental mammals in the precocious ossification of the bones surrounding the oral cavity, but share with monotremes an extended period of neurocranial osteogenesis. © 1993 Wiley-Liss, Inc.     

Testosterone Control of Male-Type Sexual Behavior in the Tammar Wallaby (Macropus eugenii)

In the tammar wallaby,Macropus eugenii,the expression of male-type sexual behavior is apparently determined by the activating effects of testicular hormones in adulthood. The incidence of male-type copulatory behavior and sexual checking behavior was compared in intact (control) males, control females, testosterone-treated females, and three groups of males castrated either postnatally (24–26 days of age), prepubertally (14.5 months of age), or in adulthood. All three groups of castrated male wallabies showed a very low incidence of male sexual behavior in adult life, comparable to that shown by the untreated females. Adult female wallabies with 100-mg testosterone implants showed a high incidence of male sexual behavior which was indistinguishable from that shown by intact males. The results suggest that sex differences in male-type behavior in the tammar wallaby are due to short-term inductive effects of testosterone acting on a sexually indifferent brain. There is no evidence of any long-term organizational effects of testosterone acting in fetal or neonatal life on the neural pathways controlling male-type sex behavior in this marsupial mammal.     

Embryo implantation and proteinase activities in a marsupial (Macropus eugenii)

Summary Embryo implantation remains superficial (epithelio-chorial type) in most marsupials including the Macropodidae, but does involve formation of specialized contact zones of the trophoblast with the uterine epithelium. Since in eutherian mammals proteinases appear to play a central role in implantation-initiation mechanisms, a systematic histochemical investigation of proteinase patterns as related to implantation was performed in the tammar wallaby, Macropus eugenii (Macropodidae).Tammar uteri with embryos were collected at diapause and at days 7, 17, 18, 19, 20, 21 and 26 of the 27-day gestational period. Proteinase patterns were studied using a sensitive histochemical gelatin-substrate-film test previously optimized for the detection of trophoblast-dependent proteinase (blastolemmase) in the rabbit. Proteinase patterns were correlated with light-microscopical morphology of the processes of shedding of the extracellular embryo coverings (shell membrane) and attachment of the trophoblast to the uterine epithelium.At acid pH values an intracellular proteinase is detected in yolk sac endoderm and trophoblast as well as in endometrial glands and certain stromal cells. This enzyme is proposed to be a cathepsin indicating high catabolic activity connected particularly with protein transport from the endometrium into the yolk sac. Peak activity is found in the avascular (bilaminar) yolk sac at the phase when contact with the endometrium is being established.A particularly interesting proteinase active at alkaline pH values is detected in the trophoblast-endoderm complex. This enzyme appears to be extruded into the interface between trophoblast and uterine epithelium where it shows maximal activity for only approximately one day, around day (18-)19, exclusively in the bilaminar (avascular) yolk sac. The activity is correlated with the process of shedding of the extracellular embryo coverings (shell membrane) and of subsequent attachment of the trophoblast to the uterine epithelium, in the bilaminar but not the trilaminar (vascular) yolk-sac region. This is the first report on an extracellular (alkaline) proteinase activity possibly serving a specific function in embryo implantation in a marsupial.Abreviations BYS bilaminar (avascular) yolk sac membrane = bilaminar omphalopleure - dp.c. days post coitum - d RPY days after removal of pouch young - TYS trilaminar (vascular) yolk sac membrane = trilaminar omphalopleure Preliminary reports on portions of these investigations were presented at the 14th Annual Meeting of the Society for the Study of Reproduction 1981 (Biol Reprod 24 Suppl 1, p 78 A, 1981) and at the 3. Arbeitstagung der Anatomischen Gesellschaft 1982 (Anat Anz 153, 268, 1983)     

Structure of the pars distalis in the adult tammar wallaby (Macropus eugenii)

An immunohistochemical, light- and electron-microscopial study was made of the pars distalis in adult tammar wallabies (Macropus eugenii). The pars distalis of this marsupial mammal was divided into three regions, based on the distribution of cell types within the gland. Somatotropic, mammotropic, luteotropic, folliculotropic, corticotropic and thyrotropic cells were identified on the basis of their immunohistochemistry, cytology and ultrastructure. Non-granulated (folliculo-stellate) cells, identified in electron micrographs, were found throughout the pars distalis. Somatotropic cells were predominant in the posterior pars distalis in all animals examined. In the single male specimen and in the non-lactating females examined, small numbers of apparently inactive mammotropic cells were scattered throughout the pars distalis; the same cell type was apparently active and present in considerable numbers in lactating females. Only one morphological type of gonadotropic cell was evident; these cells were scattered throughout the pars distalis, but in largest numbers in the median region. Small numbers of thyrotropic cells were found, most commonly in the anterior pars distalis. Corticotrops were also observed in moderate numbers, predominantly in the anterior regions of the pars distalis.     

Cyclic AMP-dependent tyrosine phosphorylation in tammar wallaby (Macropus eugenii) spermatozoa

Despite considerable advances in our understanding of the molecular mechanisms regulating eutherian sperm function, there is a paucity of such knowledge for the Metatheria. In eutherian spermatozoa, the attainment of functional competence is associated with a redox-regulated, cAMP-mediated tyrosine phosphorylation cascade, activated during capacitation. In this report we investigate whether tammar wallaby (Macropus eugenii) spermatozoa possess a similar signal transduction pathway. Western blot analysis of phosphotyrosine expression in caudal and ejaculated populations of tammar spermatozoa revealed that elevation of intracellular cAMP levels, but not exposure to oxidants or NADPH, induced a dramatic increase in the overall level of tyrosine phosphorylation. Washed, ejaculated spermatozoa exhibited more pronounced increases in tyrosine phosphorylation than unwashed sperm populations. Localisation of tyrosine phosphorylation by immunocytochemistry showed that phosphotyrosine residues were principally located along the tammar sperm flagellum, and occasionally at a small region of the sperm head, adjacent to the acrosome. Associated with the tyrosine phosphorylation of tammar spermatozoa, was a change in sperm head conformation to a T-shaped orientation, further implying the importance of these pathways to normal tammar sperm function. Redox activity, as detected by lucigenin-dependent chemiluminescence, was stimulated by NADPH in caudal sperm preparations but not ejaculated spermatozoa. However, neither sperm population responded to treatment with NADPH with changes in intracellular cAMP or tyrosine phosphorylation. In conclusion, tammar spermatozoa possess the same cAMP-mediated, tyrosine phosphorylation-dependent signal transduction cascade that has been associated with capacitation in eutherian spermatozoa. However in Metatherian spermatozoa we could find no evidence that this pathway was redox regulated.     

Peace-Making in Marsupials: The First Study in the Red-Necked Wallaby (Macropus rufogriseus)

The issue of reconciliation has been widely investigated in many eutherian mammal species. Nevertheless, no data are available for marsupial mammals. Indeed, the majority of reports focus on group dynamics from an ecological and reproductive perspective, but no study has investigated them from a social point of view. We observed the red-necked wallaby colony (Macropus rufogriseus) hosted at the Tierparc Zoo Berlin (Germany) and collected data on aggressive and post-conflict interactions between group members. We found that the phenomenon of reconciliation is present in the study species (mean group CCT 27.40% ±8.89% SE). Therefore, we demonstrated, for the first time, the occurrence of reconciliation in a gregarious marsupial mammal. Post-conflict reunion was not affected by the relationship quality between individuals (friendship or kinship) but it was fine-tuned according to the aggression intensity. For example, low intensity conflicts were reconciled whereas high intensity ones were not. Reconciliation reduced anxiety-related scratching in both of the former opponents and limited further attacks towards the victim during the post-conflict period. These findings suggest that the red-necked wallaby, like many eutherian species, can evaluate the costs of reconciliation and engage in peace-making behavior in the right contexts, in order to maximize its pay-offs.     

Characterization and expression of Peroxiredoxin 1 in the neonatal tammar wallaby (Macropus eugenii)

Peroxiredoxin 1 (PRDX1) is a ubiquitously expressed antioxidant with vital roles in basal metabolic functions. In addition PRDX1 is involved in cell differentiation and proliferation, apoptosis and innate immunity. In this study, we have characterized PRDX1 from the tammar wallaby (Macropus eugenii). Tammar PRDX1 has high conservation of functional residues and motifs, and demonstrates a close homology with eutherian and vertebrate orthologues. Stimulation of adult tammar leukocytes with lipopolysaccharide and lipoteichoic acid suggests a role for PRDX1 in innate immune defences. PRDX1 expression in the organs of tammar pouch young was mildly elevated early in life possibly reflecting its role in basal metabolic processes. Later increases in PRDX1 expression correlated with functional maturation of several immune organs or with preparation for increased oxidative stress of emergence. The findings of the study are reflections of the complex integrated roles that PRDX1 has in regulation of oxidative stress, apoptosis, cell differentiation and proliferation, and innate immunity.     

Control of jaw movements in two species of macropodines (Macropus eugenii and Macropus rufus).

The masticatory motor patterns of three tammar wallabies and two red kangaroos were determined by analyzing the pattern of electromyographic (EMG) activity of the jaw adductors and correlating it with lower jaw movements, as recorded by digital video and videoradiography. Transverse jaw movements were limited by the width of the upper incisal arcade. Molars engaged in food breakdown during two distinct occlusal phases characterized by abrupt changes in the direction of working-side hemimandible movement. Separate orthal (Phase I) and transverse (Phase II) trajectories were observed. The working-side lower jaw initially was drawn laterally by the balancing-side medial pterygoid and then orthally by overlapping activity in the balancing- and working-side temporalis and the balancing-side superficial masseter and medial pterygoid. Transverse movement occurred principally via the working-side medial pterygoid and superficial masseter. This pattern contrasted to that of placental herbivores, which are known to break down food when they move the working-side lower jaw transversely along a relatively longer linear path without changing direction during the power stroke. The placental trajectory results from overlapping activity in the working- and balancing-side adductor muscles, suggesting that macropods and placental herbivores have modified the primitive masticatory motor pattern in different ways.