Two stages of increased IgA transfer during lactation in the marsupial, trichosurus vulpecula (Brushtail possum)

The polymeric Ig receptor (pIgR) and J chain molecules are involved in the transfer of IgA across the mammary gland epithelia into milk. The J chain binds two IgA molecules to form dimeric IgA, and the pIgR transports this complex through epithelial cells. We report here the cloning of the first marsupial homologues for the pIgR and J chain from the brushtail possum. Marsupial young are born after a short gestation and are less developed than eutherian newborn. The pouch young is completely dependent on milk as its sole source of nutrition during early lactation and this phase can be considered to be equivalent to an external gestation. Two periods of increased expression of pIgR, J chain, and IgA heavy chain mRNAs were observed in the mammary gland during lactation. The first occurs for a brief period after birth of the pouch young and is likely to reflect IgA transfer via the colostrum. The second period of increased expression, which is unique to marsupials, occurs after the early lactation period and just before young exit the pouch. We propose that this represents a second colostral-like phase at the end of the external gestation.     

Temperature gradient from the urogenital sinus to the pouch in the pregnant marsupial quoll, Dasyurus hallucatus

1. The mechanisms utilised by the newborn quoll to move from the uterus to the teat within the pouch are unknown. The ability to sense gravity and odour have been suggested and it is possible that temperature may also assist the young in this migration.

2. An increasing temperature gradient was observed from the sinus at 28.98 °C increasing to 29.38 °C on the skin between the sinus and the pouch and further increasing to 30.96 °C within the pouch. This temperature gradient was not as apparent during lactation.

3. Hairs may also play an important role in allowing the newborn to leave the gelatinous material emanating from the uterus and travel to the pouch. The hairs form a tunnel between the sinus and the pouch and may assist the young in the moving from uterus to the pouch.     

PATTERNS OF PARENTAL CARE AND PARENTAL INVESTMENT IN MARSUPIALS

I. Information on growth, development and care of young has been assembled for 62 species of marsupial. 2. During gestation, development of the marsupial embryo proceeds only so far as to allow the neonate to make its way from the urogenital opening to the mammary area on the abdomen of the female where it attaches to a teat. Specific structural adaptations keep the neonate firmly attached to the teat for at least the first month after birth. 3. Six types of pouch are distinguished ranging from lateral folds of skin, which do not cover the mammary area or enclose the developing young, to a fold of skin that covers the mammary area and forms a deep pouch, completely enclosing the developing young. 4. Although the young is very small at birth and birth is rapid, specific changes in the behaviour of females occur around the time of birth, and a specific birth position is adopted. 5. The time at which marsupial young leave the pouch cannot be equated with birth in eutherians, because of the considerable variations in the type of pouch and in patterns of parental care. From a consideration of the functional development of the young in the pouch, it is suggested that the nearest equivalent to eutherian birth is the time at which the marsupial young achieves homeostasis, when it is well furred and endothermic. 6. Maternal behaviour is influenced by the type of pouch. In all species, the mother keeps the young and the pouch clean by licking, especially when the young are wholly within the pouch or attached to the abdomen. In species with reduced pouches where young are left in a nest at an early stage of development, maternal behaviour includes nest building, defence, and retrieving and carrying the young. These functions are performed by the pouch itself in species with large deep pouches in which the young is carried for a much larger part of its development, and other specific maternal behaviours are infrequent. 7. The patterns of parental care are reviewed over all families of marsupial. Not all members of a family have the same pattern of parental care, which appears to be influenced by many factors including body size, type of pouch, diet, litter size and other aspects of life history strategy. 8. Three patterns of parental care are distinguished: (A) As soon as young begin to release the teat they are no longer carried by the mother, and are left in a nest when still barely furred, ectothermic and before the eyes open. This pattern is found in species with large litter size and a pouch reduced or absent, e.g. some Dasyuridae and some Didelphidae. (B) Young remain in the pouch after they begin to release the teat but are left in a nest, at a later stage of development than in A, when well furred, endothermic and with eyes open. After first pouch exit, there is generally a period when young return to the pouch from time to time. This pattern is found in species with well developed pouches and litters of I or > 1 e.g. Peramelidae, some Didelphidae. (C) Young remain in the pouch after they begin to release the teat. At first pouch exit, the young is well furred and endothermic, and leaves the pouch only for brief periods, gradually spending more time out until permanent pouch exit. It is not usually left in a nest. This pattern is found in species with well developed pouches and litters of one, e.g. Macropodidae. 9. Pattern A is seen particularly in the smaller species in any family, where large litter size means that by the time young release the teat, the litter is about 50% of maternal body weight and a considerable burden. In such species, young are left in a nest as soon as possible. In larger species with patterns B or C, litter size is smaller, and by the time they are no longer carried by the female, the litter is still only 20% of maternal weight. 10. Whatever the pattern of parental care, mortality from birth to permanent pouch exit is not unusually high in marsupials in comparison with eutherians. 11. I suggest that the presence of the pouch and the associated patterns of parental care are important determinants of social organization in marsupials. For much of the period of dependence, the young is small, attached to a teat or in a pouch. The male can make little contribution to parental care, and there is little room for improvement in the care of young in complex social groups. In most species, the female on her own is sufficient caretaker. The male is most likely to increase his own biological fitness by going off to mate again and leaving the female to raise his offspring. 12. Patterns of energy expenditure on offspring by female marsupials were assessed throughout the development of young. Investment before birth was assessed by weight of the neonate, during development by growth rate and the time for which the young was carried (pouch life), and total investment by weight of young at weaning and time from birth to weaning. Regression of measures of investment against maternal body weight allowed comparison of investment in animals of different size. 13. Investment in young before birth is very small. Neonatal marsupials range in size from 0·01 to 1 g, and the largest is less than 0·2 % of the size of the mother. Larger mothers produce larger young which are smaller relative to the mother than are the young of smaller species. Individual young in the family Dasyuridae are particularly small. 14. Growth rates in g/d were calculated over the period from permanent pouch exit to weaning. There is a very close correlation between growth rate and maternal body weight - that of litters increases as the 0·78 power of body weight. During this period the growth rate of individuals is comparable with that of eutherian young during lactation, and in litters it is higher still, suggesting that the difference in patterns of growth are not due to the lower metabolic rate of marsupials. As in eutherians there is considerable individual variation in growth rate; it is very high in litters of small dasyurids, which have individual rates comparable to those of larger species. Young of the family Peramelidae grow and develop rapidly; those of the arboreal folivore Phascolarctos do both slowly. I 5. Pouch life, the period for which the young is carried by the mother, increases with body size; as expected, species with pattern A parental care have shorter pouch lives than species of the same size with patterns B or C, reflecting the early stage of development at which young are left in the nest in pattern A. 16. Time from birth to weaning is also longer in larger species. There is a close relationship of age at weaning with maternal weight, with some significant exceptions. For their size, the family Peramelidae have a very short time from birth to weaning, and the time from pouch exit to weaning is particularly short. Many arboreal species have longer periods of dependence than expected from their size. 17. The weight at weaning of individual young is closely related to MBW^0·71, but the weight of one young relative to maternal body weight shows no trend with size, and ranges from 25–61 %, with a mean of 42 %. 18. Parental Investment, as measured by the function Wt. of litter at weaning × 100/MBW, decreases with increasing size of mother as MBW^0·28. The highest levels of investment are found in very small species. In many small species of the family Dasyuridae, a litter at weaning is > 300% MBW. By contrast, investment in the family Peramelidae is low - at weaning a litter of three is about 50% MBW, comparable with investment in a single young of the family Macropodidae. 19. The evolution of patterns of parental care and investment appears to follow three main lines: (1) Species with large litter size, high levels of investment in litters and in individual young. Investment is directed to growth and not to carrying the young in the pouch, since young are left in a nest at an early stage. Typical of this group is the family Dasyuridae, in which many species make few reproductive attempts per year. (2) Species with litters of more than one, low levels of investment in litters and in individuals, but rapid growth and development of young. Because of the small relative size of young they are carried in the pouch for a large part of the period from birth to weaning. This pattern is shown by the family Peramelidae, and seen as an adaptation to rapid and repeated reproduction in an environment with an extended favourable season. (3) Species with small litter size, lower total investment, but investment in individual young is not low, and investment in carrying young to an advanced stage of development is high. Patterns of this type are found in the Diprotodonta, with extreme development in the Macropodidae. 20. Many of the measures of investment have been expressed as a power function of maternal body weight. The exponents of body weight in these functions are such as to suggest that an important underlying variable is metabolic rate. 21. It has been suggested elsewhere that the marsupial mode of reproduction evolved as an adaptation to environmental uncertainty, in that it allows a reproductive attempt to be abandoned at any time much more readily than in eutherians, thereby increasing the likelihood that a female will survive to reproduce again. I consider this suggestion in the light of patterns of parental investment. For small, short-lived species, any reproductive attempt represents a substantial part of its lifetime reproductive output. Investment in any one reproductive attempt is high, and the cost of replacing an abandoned attempt is so high that it seems unlikely that the desertion of offspring would be an important reproductive strategy in small ancestral marsupials, although it may be an important response to environmental uncertainty in certain large modern macropodids.     

Reduction in the number of young during pouch-life in a small marsupial

The honey possum Tarsipes rostratus , a shrew-sized marsupial endemic to south-western Australia, feeds exclusively upon nectar and pollen. Tarsipes has fewer pouch-young, which remain in the pouch longer, than the young of sympatric insect-feeding marsupials of comparable size. Tarsipes pouch-young were recorded in every month of the year. Of all adult females caught, 68% had pouch-young and 36% of the remainder had recently carried young. This continual reproduction is offset by an average lifespan of only about one year.
Although Tarsipes have four teats, only very few of the largest females reared four young to independence. The number of pouch-young was inversely related to their size, changing, on average, from 3.6 young immediately after birth to 2.4 young at pouch-exit. Larger females gave birth to larger litters than smaller females and appeared more successful in rearing them. Variation in the initial litter size, and any subsequent reduction in litter size, might allow females to optimize offspring production during their short lifetime in relation to an unpredictable and specialized food supply.     

Virilization of the male pouch young of the tammar wallaby does not appear to be mediated by plasma testosterone or dihydrotestosterone.

Virilization of the male urogenital tract of all mammals, including marsupials, is mediated by androgenic hormones secreted by the testes. We have previously demonstrated profound sexual dimorphism in the concentrations of gonadal androgens in pouch young of the tammar wallaby Macropus eugenii during the interval when the urogenital sinus virilizes. To provide insight into the mechanisms by which androgens are transported from the testes to the target tissues, we measured testosterone and dihydrotestosterone in plasma pools from tammar pouch young from the day of birth to Day 150. Plasma testosterone levels were measurable (0.5-2 ng/ml) at all times studied, but there were no differences between males and females. These low concentrations of plasma testosterone appear to be derived from the adrenal glands and not the testes. Plasma dihydrotestosterone levels in plasma pools from these animals were also low and not sexually dimorphic. We conclude that virilization of the male urogenital tract cannot be explained by the usual transport of testosterone or dihydrotestosterone in plasma but may be mediated by the direct delivery of androgens to the urogenital tract via the Wolffian ducts. Alternatively, circulating prohormones may be converted to androgens in target tissues.     

Identification of novel major histocompatibility complex class I sequences in a marsupial,the brushtail possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The major histocompatibility complex (MHC) is an essential part of the vertebrate immune response. MHC genes may be classified as classical, non-classical or non-functional pseudogenes. We have investigated the diversity of class I MHC genes in the brushtail possum, a marsupial native to Australia and an introduced pest in New Zealand. The MHC of marsupials is poorly characterised compared to eutherian mammal species. Comparisons between marsupials and eutherians may enhance understanding of the evolution and functions of this important genetic region. We found a high level of diversity in possum class I MHC genes. Twenty novel sequences were identified using polymerase chain reaction (PCR) primers designed from existing marsupial class I MHC genes. Eleven of these sequences shared a high level of homology with the only previously identified possum MHC class I gene TrvuUB and appear to be alleles at a single locus. Another seven sequences are also similar to TrvuUB but have frame-shift mutations or stop codons early in their sequence, suggesting they are non-functional alleles of a pseudogene locus. The remaining sequences are highly divergent from other possum sequences and clusters with American marsupials in phylogenetic analysis, indicating they may have changed little since the separation of Australian and American marsupials.     

Prenatal bias in sex ratios in a marsupial,Antechinus agilis.

Biased sex ratios of young of birds and mammals clearly occur and may have an adaptive significance, but we rarely know the stage at which a bias is generated, or the mechanism. If a bias is generated prior to birth, studies of marsupials may be insightful because gestation is short and neonates are relatively undifferentiated. This study investigated whether biased sex ratios in Antechinus agilis are generated in the brief period between birth and the attachment of young to the mother''s teats. When all young born, or just pouch young, or supernumerary young were considered, litters were strongly biased towards females (0.32 males), and there was no significant difference across the groups, so a bias is generated before birth in this species. Evidence from counts of corpora lutea suggests that embryo loss during gestation cannot account fully for the level of bias observed. Therefore, prefertilization mechanisms must contribute to the generation of sex-biased litters in this marsupial.     

A molecular and evolutionary study of the beta-globin gene family of the Australian marsupial Sminthopsis crassicaudata

Beta-globin gene families in eutherians (placental mammals) consist of a set of four or more developmentally regulated genes which are closely linked and, in general, arranged in the order 5'-embryonic/fetal genes- adult genes-3'. This cluster of genes is proposed to have arisen by tandem duplication of ancestral beta-globin genes, with the first duplication occurring 200 to 155 MYBP just prior to a period in mammalian evolution when eutherians and marsupials diverged from a common ancestor. In this paper we trace the evolutionary history of the beta-globin gene family back to the origins of these mammals by molecular characterization of the beta-globin gene family of the Australian marsupial Sminthopsis crassicaudata. Using Southern and restriction analysis of total genomic DNA and bacteriophage clones of beta-like globin genes, we provide evidence that just two functional beta-like globin genes exist in this marsupial, including one embryonic- expressed gene (S.c-epsilon) and one adult-expressed gene (S.c-beta), linked in the order 5'-epsilon-beta-3'. The entire DNA sequence of the adult beta-globin gene is reported and shown to be orthologous to the adult beta-globin genes of the North American marsupial Didelphis virginiana and eutherian mammals. These results, together with results from a phylogenetic analysis of mammalian beta-like globin genes, confirm the hypothesis that a two-gene cluster, containing an embryonic- and an adult-expressed beta-like globin gene, existed in the most recent common ancester of marsupials and eutherians. Northern analysis of total RNA isolated from embryos and neonatals indicates that a switch from embryonic to adult gene expression occurs at the time of birth, coinciding with the transfer of the marsupial from a uterus to a pouch environment.      

Urogenital vasculature and local steroid concentrations in the uterine branch of the ovarian vein of the female tammar wallaby (Macropus eugenii)

The urogenital vasculature of the tammar comprises 4 major paired arteries and veins: the ovarian, the cranial urogenital, the caudal urogenital and the internal pudendal artery and vein. The ovarian artery and vein and their uterine branches which supply the ovary, oviduct and uterus, ramify extensively. Each anterior urogenital artery and vein supplies the caudal regions of the ipsilateral uterus, lateral and median vagina and cranial parts of the urogenital sinus. The caudal urogenital arteries and veins supply the urogenital sinus and caudal regions of the bladder. The internal pudendal artery and vein vascularize the cloacal region, with some anastomoses with branches of the external pudendal vessels. Anastomoses connect the uterine branch of the ovarian artery with the uterine branch of the cranial urogenital and cranial branches of the caudal urogenital arteries, and connect the caudal urogenital and the internal pudendal arteries. Anastomotic connections between the left and right arterial supply also occur across the midline of the cervical regions of the uteri and the anterior lateral vaginae. Similar connections are seen in the venous system. The uterine branch of the ovarian artery ramifies extensively very close to the ovary, giving a plexiform arrangement with the ovarian veins, and also with the uterine venous system on the lateral side of each uterus. This plexiform structure provides an anatomical arrangement which could allow a local transfer of ovarian hormones from ovarian vein into the uterine arterial supply, and thence to the ipsilateral uterus. Progesterone concentrations in plasma from the mesometrial side of the uterine branch of the ovarian vein are markedly higher than in tail vein plasma, especially during the 'Day 5 peak' early in pregnancy, and also at full term. There is also a marked decrease in progesterone concentration from all sites immediately before birth as previously reported for peripheral plasma. These results support the suggestion of a countercurrent transfer mechanism, at least for progesterone, and possibly other hormones, between the ovarian vein and uterine artery. Such a local transfer could explain the different morphological responses of the endometria of the two adjacent uteri during pregnancy in macropodid marsupial species.     

Development of the urinary system of the marsupial native cat Dasyurus hallucatus.

The development of the mesonephros and metanephros was examined in the marsupial Northern native cat, Dasyurus hallucatus, from birth through to the end of lactation. The mesonephros was present at birth, reached a maximum volume 11 days after birth and had regressed completely by day 30. The metanephros was present on day 2; glomeruli were first seen on day 8, and nephrogenesis continued until day 89 post partum. The newborn native cat is at a very primitive stage of development compared to other marsupials. However, at weaning, like other marsupial species, the native cat has developed a fully morphological and functional urinary system.     

Artificial insemination in marsupials

Assisted breeding technology (ART), including artificial insemination (AI), has the potential to advance the conservation and welfare of marsupials. Many of the challenges facing AI and ART for marsupials are shared with other wild species. However, the marsupial mode of reproduction and development also poses unique challenges and opportunities. For the vast majority of marsupials, there is a dearth of knowledge regarding basic reproductive biology to guide an AI strategy. For threatened or endangered species, only the most basic reproductive information is available in most cases, if at all. Artificial insemination has been used to produce viable young in two marsupial species, the koala and tammar wallaby. However, in these species the timing of ovulation can be predicted with considerably more confidence than in any other marsupial. In a limited number of other marsupials, such precise timing of ovulation has only been achieved using hormonal treatment leading to conception but not live young. A unique marsupial ART strategy which has been shown to have promise is cross-fostering; the transfer of pouch young of a threatened species to the pouches of foster mothers of a common related species as a means to increase productivity. For the foreseeable future, except for a few highly iconic or well studied species, there is unlikely to be sufficient reproductive information on which to base AI. However, if more generic approaches can be developed; such as ICSI (to generate embryos) and female synchronization (to provide oocyte donors or embryo recipients), then the prospects for broader application of AI/ART to marsupials are promising.     

Birth of pouch young after artificial insemination in the tammar wallaby (Macropus eugenii)

Timing of artificial insemination (AI) in marsupials is critical because fertilization must occur before mucin coats the oocyte during passage through the oviduct. In this study, timing and the site of insemination were examined to develop AI in the tammar wallaby (Macropus eugenii). Birth and postpartum (p.p.) estrus was synchronized in 46 females. Epididymal spermatozoa (n=4) or semen collected by electroejaculation (n=42) were inseminated early (4-21 h p.p.) into the urogenital sinus (n=7), the anterior vaginal culs de sac (n=7), the uterus by transcervical catheter (n=5), or the uterus by injection (intrauterine artificial insemination, IUAI) (n=5). A further 16 females were inseminated late (19-48 h p.p.) by IUAI. All females were monitored for birth. A third group of six females was inseminated late (21-54 h p.p.) by IUAI and 0.4-6.6 h later, sperm had reached the oviduct in all animals. In total, an oocyte to which spermatozoa were attached was recovered and two young were born after IUAI using epididymal (n=1) or electroejaculated (n=2) spermatozoa, but no young resulted from insemination at other sites. Two females were successfully inseminated at 43 and 47 h p.p., later than most other animals, and the third was inseminated much earlier (18 h p.p.) but with highly motile spermatozoa. These young represent the first macropodids born by AI and the first marsupials conceived using epididymal spermatozoa.     

Mitochondrial genomes of a bandicoot and a brushtail possum confirm the monophyly of australidelphian marsupials.

Recent molecular analyses suggest that the position of bandicoots is the major difficulty in determining the root of the tree of extant marsupials. To resolve this, we analyse mitochondrial genome sequences of a bandicoot (Isoodon macrourus) and a brushtail possum (Trichosurus vulpecula) together with the previously available marsupial mitochondrial genomes, the Virginia opossum (Didelphis virginiana) and the wallaroo (Macropus robustus). Analyses of mitochondrial protein-coding and RNA genes strongly support the bandicoot as sister to the wallaroo and the brushtail possum. This result, combined with other recent molecular analyses, confirms the monophyly of Australidelphia (Australasian marsupials plus Dromiciops from South America). Further, RY coding was found to nullify AGCT coding nucleotide composition bias.     

Non‐invasive placentation in the marsupials Macropus eugenii (Macropodidae) and Trichosurus vulpecula (Phalangeridae) involves redistribution of uterine Desmoglein‐2

In mammalian pregnancy, the uterus is remodeled to become receptive to embryonic implantation. Since non‐invasive placentation in marsupials is likely derived from invasive placentation, and is underpinned by intra‐uterine conflict between mother and embryo, species with non‐invasive placentation may employ a variety of molecular mechanisms to maintain an intact uterine epithelium and to prevent embryonic invasion. Identifying such modifications to the uterine epithelium of marsupial species with non‐invasive placentation is key to understanding how conflict is mediated during pregnancy in different mammalian groups. Desmoglein‐2, involved in maintaining lateral cell–cell adhesion of the uterine epithelium, is redistributed before implantation to facilitate embryo invasion in mammals with invasive placentation. We identified localization patterns of this cell adhesion molecule throughout pregnancy in two marsupial species with non‐invasive placentation, the tammar wallaby (Macropus eugenii; Macropodidae), and the brushtail possum (Trichosurus vulpecula; Phalangeridae). Interestingly, Desmoglein‐2 redistribution also occurs in both M. eugenii and T. vulpecula, suggesting that cell adhesion, and thus integrity of the uterine epithelium, is reduced during implantation regardless of placental type, and may be an important component of uterine remodeling. Desmoglein‐2 also localizes to the mesenchymal stromal cells of M. eugenii and to epithelial cell nuclei in T. vulpecula, suggesting its involvement in cellular processes that are independent of adhesion and may compensate for reduced lateral adhesion in the uterine epithelium. We conclude that non‐invasive placentation in marsupials involves diverse and complementary strategies to maintain an intact epithelial barrier.     

The transfer of thyroxine from the mother to the young of the marsupials,the bandicoot (Isoodon macrourus) and the possum (Trichosurus vulpecula)

• 1.1. The young of the bandicoot and possum are of similar weight at birth (200–250 mg) but by day 50 post-partum have weights of 100 and 20 g, respectively. The aim of this study was to determine whether the disparate growth rate was associated with a difference in the transfer of thyroxine from mother to young.
• 2.2. Radioactive thyroxine was injected intramuscularly into the lactating female and 24 hr later a blood sample was obtained from the mother and the pouch young removed. The amount of radioactive thyroxine remaining in the blood and within the young was determined.
• 3.3. The data obtained indicated that milk is a source of thyroid hormones in early pouch life.
• 4.4. The hypothesis that greater quantities of thyroxine are transferred in the milk of bandicoots than that of brushtail possums is not supported.

     

The hormonal control of birth behavior in the gray short-tailed opossum (Monodelphis domestica)

In all major groups of Australian marsupials, prostaglandin F2alpha (PGF) or oxytocin injection initiates birth behavior in adult females, adult males and pouch young. Because inhibitors of PGF synthesis block this initiation, oxytocin may activate birth behavior via the stimulation of PGF synthesis. In this study, the role of PGF and oxytocin in the activation of birth behavior was examined in an American marsupial, the gray short-tailed opossum (Monodelphis domestica). Adult male and female gray opossums were given PGF, oxytocin, or saline (control) before behavioral observation. On the next day, the animals in the oxytocin group were injected with the PGF inhibitor flunixin meglumide (Finadyne, Schering Corp., U.S.A.) before oxytocin reinjection and behavioral observation. Both males and females showed birth behavior in response to PGF but only females responded to oxytocin. There was no significant difference in the latency of response of females to oxytocin alone versus response to oxytocin after receipt of the PGF inhibitor. These results suggest that, in contrast to Australian species, in this American marsupial, oxytocin initiates birth behavior only in females and does not operate via stimulation of prostaglandin secretion.     

The ultrastructure of the lung of two newborn marsupial species,the northern native cat,Dasyurus hallucatus,and the brushtail possum,Trichosurus vulpecula

Summary The lungs of newborn northern native cats, Dasyurus hallucatus and newborn brushtail possums, Trichosurus vulpecula were examined by both light and electron microscopy. The native cat has a birth weight of 18 mg after a gestation of about 21 days, whereas the brushtail possum weights 200 mg at birth and has a gestation period of 17.5 days. The lungs of the native cat are two large respiratory sacs, with a respiratory lining of squamous cells and surfactant-secreting cells. The capillaries are located within the connective tissue just below this respiratory epithelium. The visceral covering of the lung is formed by squamous cells. The lungs of the possum are composed of numerous large respiratory sacs which are separated by connective tissue septa in which the capillaries are located. The sacs, as in other species, are lined with squamous cells and surfactant secreting cells. It is proposed that the structure of the lung of the newborn marsupial is related more to the size of the newborn rather than to the length of the gestation period.     

Sex determination in marsupials: evidence for a marsupial-eutherian dichotomy

In this paper, we review briefly the current state of knowledge about sexual differentiation in eutherian mammals, and then describe the situation in detail in two marsupial species: the North American opossum and the tammar wallaby. The conventional explanation for the genesis of all male somatic sexual dimorphisms in mammals is that they are a consequence of the systemic action of testicular hormones. In the absence of testes, the embryo will develop a female phenotype. We present evidence for the tammar wallaby that calls into question the universal applicability of this hormonal theory of mammalian sexual differentiation. We have shown that extensive somatic sexual dimorphisms precede by many days the first morphological evidence of testicular formation, which does not occur until around the third day of pouch life. Male foetuses, and pouch young on the day of birth, already have a well-developed gubernaculum and processus vaginalis, paired scrotal anlagen, and a complete absence of mammary anlagen, whereas female foetuses and newborn pouch young have a poorly developed gubernaculum and processus vaginalis, no scrotal anlagen, and well-developed mammary anlagen. Because it seems unlikely that the male gonad could begin hormone secretion until after the Sertoli and Leydig cells are developed, our results strongly suggest that some sexually dimorphic somatic characteristics develop autonomously, depending on their genotype rather than the hormonal environment to which they are exposed. We have been able to confirm the hormonal independence of the scrotum, pouch and mammary gland by administering testosterone propionate daily by mouth to female pouch young from the day of birth; although the Wolffian duct was hyperstimulated, there was no sign of scrotal development, or pouch or mammary inhibition. When male pouch young were treated with oestradiol benzoate in a similar fashion, there was hyperstimulation of the Müllerian duct and inhibition of testicular migration and development, but no sign of scrotal inhibition or pouch or mammary development. Our results in the tammar wallaby are consistent with the earlier studies on the opossum, whose significance was not appreciated at the time. Further evidence in support of this hormonal independence comes from earlier studies of spontaneously occurring intersexes in several species of marsupial, including the opossum and the tammar wallaby. An XXY individual had intra-abdominal testes and complete masculinization of the male reproductive tract internally, but externally there was a pouch and mammary glands and no scrotum.(ABSTRACT TRUNCATED AT 400 WORDS)