Cytoskeletal organization in the oocyte,zygote, and early cleaving embryo of the stripe-faced dunnart (Sminthopsis macroura)

Ovulation occurs in Sminthopsis macroura approximately 160 hr after administration of 1.3 IU PMSG, and yields significantly more oocytes than does spontaneous ovulation (P = 0.001). Germinal vesicle (GV)-stage oocytes have a thin cortical rim of microfilaments, which is disrupted by exposure to cytochalasin D. After GV breakdown, the first meiotic spindle forms subcortically and parallel to the oolemma. It rotates during anaphase and telophase to extrude the first polar body. This rotation is associated with a local cortical concentration of microfilaments, which is extruded in the first polar body. The second meiotic spindle is orthogonal to the surface, and extrusion of the second polar body is not associated with obvious local changes in cortical actin, resulting in a polar body containing little polymerized actin. The sites of second polar body emission and sperm entry are always in the half of the oocyte opposite the concentrating yolk mass, and are within 60° of each other in most oocytes. During the concentration and eccentric movement of the yolk, microfilaments condense around it. During yolk expulsion, these microfilaments become continuous with those located subcortically. During early cleavage, the cytocortex of the zygote, but not of the extruded yolk mass, stains heavily for polymerised actin. Multiple sites of pericentriolar material are detectable in the cytoplasm of some secondary unfertilized oocytes which, in the presence of taxol, generate large cytasters and pseudospindle structures. After fertilization, a large aster is formed in association with the sperm entry point and serves as the center of an extensive cytoplasmic network of microtubules which surrounds but does not enter the yolk mass. Taxol treatment generates small cytasters within this meshwork and promotes selective stabilization of some periyolk microtubules opposite to the sperm aster. © 1995 Wiley-Liss, Inc.     

The effect of temperature on the pattern of torpor in a marsupial hibernator

Physiological variables of torpor are strongly temperature dependent in placental hibernators. This study investigated how changes in air temperature affect the duration of torpor bouts, metabolic rate, body temperature and weight loss of the marsupial hibernator Burramys parvus (50 g) in comparison to a control group held at a constant air temperature of 2°C. The duration of torpor bouts was longest (14.0±1.0 days) and metabolic rate was lowest (0.033±0.001 ml O₂·g^-1·h^-1) at2°C. At higher air temperatures torpor bouts were significantly shorter and the metabolic rate was higher. When air temperature was reduced to 0°C, torpor bouts also shortened to 6.4±2.9 days, metabolic rate increased to about eight-fold the values at 2°C, and body temperature was maintained at the regulated minimum of 2.1±0.2°C. Because air temperature had such a strong effect on hibernation, and in particular energy expenditure, a change in climate would most likely increase winter mortality of this endangered species.Abbreviationst STP standard temperature and pressure - T _a air temperature - T _b body temperature - VO₂ rate of oxygen consumption     

Confirmation of pleisiomorphic daily torpor in mammals: the round-eared elephant shrew Macroscelides proboscideus (Macroscelidea)

The characteristics of daily torpor were measured in the round-eared elephant shrew Macroscelides proboscideus (Macroscelidea) in response to ambient temperature and food deprivation. Elephant shrews are an ancient mammal order within a superordinal African clade including hyraxes, elephants, dugongs and the aardvark. M. proboscideus only employed torpor when deprived of food; torpor did not occur under an ad libitum diet at ambient temperatures of 10, 15 and 25?°C. Torpor bout duration ranged from <1?h to ca. 18?h. The times of entry into torpor were restricted to the scotophase, despite normothermic body temperature patterns indicating a rest phase coincident with the photophase. Full arousal was always achieved within the first 3?h of the photophase. When food deprived, the onset of the rest phase, and hence torpor, advanced with respect to the experimental photoperiod. The lowest torpor body temperature measured was 9.41?°C. Daily torpor in M. proboscideus confirms a pleisiomorphic origin of daily heterothermy. Torpor facilitates risk-averse foraging behaviour in these small omnivores by overcoming long-term energy shortfalls generated by the inherent variability of food availability in their semi-arid, El Niño-afflicted habitats.     

Alterations in surface activity of pulmonary surfactant in Gould's wattled bat during rapid arousal from torpor

The small microchiropteran bat, Chalinolobus gouldii, undergoes large daily fluctuations in metabolic rate, body temperature, and breathing pattern. These alterations are accompanied by changes in surfactant composition, predominantly an increase in cholesterol relative to phospholipid during torpor. Furthermore, the surface activity changes, such that the surfactant functions most effectively at that temperature which matches the animal's activity state. Here, we examine the surface activity of surfactant from bats during arousal from torpor. Bats were housed at 24 degrees C on an 8:16h light:dark cycle and their surfactant was collected during arousal (28相似文献   

Reproductive status and torpor of the marsupial Sminthopsis crassicaudata: Effect of photoperiod

1. 1. We investigated the effects of photoperiod on the reproductive state and the occurrence and pattern of torpor in male Sminthopsis crassicaudata.

2. 2. Testes regressed when animals were exposed to a short photoperiod (L:D 8:16) and recrudesced under a long photoperiod (L:D 16:8).

3. 3. Animals entered torpor under both photoperiods and no significant differences were observed in the frequency or physiological variables of torpor of S. crassicaudata between the short and long photoperiods.

4. 4. The differences in the response to photoperiod in thermal physiology and reproduction suggest that, unlike in many rodent species, torpor and reproduction in S. crassicaudata are controlled by separate environmental cues and mechanisms.

     

Alterations in pulmonary surfactant after rapid arousal from torpor in the marsupial Sminthopsis crassicaudata.

Torpor in the dunnart, Sminthopsis crassicaudata, alters surfactant lipid composition and surface activity. Here we investigated changes in surfactant composition and surface activity over 1 h after rapid arousal from torpor (15-30 degrees C at 1 degrees C/min). We measured total phospholipid (PL), disaturated PL (DSP), and cholesterol (Chol) content of surfactant lavage and surface activity (measured at both 15 and 37 degrees C in the captive bubble surfactometer). Immediately after arousal, Chol decreased (from 4.1 +/- 0.05 to 2.8 +/- 0.3 mg/g dry lung) and reached warm-active levels by 60 min after arousal. The Chol/DSP and Chol/PL ratios both decreased to warm-active levels 5 min after arousal because PL, DSP, and the DSP/PL ratio remained elevated over the 60 min after arousal. Minimal surface tension and film compressibility at 17 mN/m at 37 degrees C both decreased 5 min after arousal, correlating with rapid changes in surfactant Chol. Therefore, changes in lipids matched changes in surface activity during the postarousal period.     

Development of thermoregulation and torpor in a marsupial: energetic and evolutionary implications

Altricial mammals and birds become endothermic at about half the size of adults and presumably would benefit energetically from entering torpor at that time. Because little is known about torpor during development in endotherms, we investigated whether after the establishment of endothermic thermoregulation (i.e. the ability to maintain a high body temperature during cold exposure), Sminthopsis macroura, a small (∼25 g) insectivorous marsupial, is capable of entering torpor and whether torpor patterns change with growth. Endothermic thermoregulation was established when the nest young reached a body mass of ∼10 g, and they were capable of entering torpor early during development at ∼10–12 g, lending some support to the view that torpor is a phylogenetically old mammalian trait. Torpor bout length shortened significantly and the minimum metabolic rate during torpor increased as juveniles approached adult size, and consequently total daily energy expenditure increased steeply with age. Relationships between total daily energy expenditure and body mass during development of S. macroura (slope ∼1.3) differed substantially from the relationship between basal metabolism and body mass in adult endotherms (slope ∼0.75) suggesting that the energy expenditure–size relationship during the development differs substantially from that in adults under thermo-neutral conditions. Our study shows that while torpor can substantially reduce energy expenditure during development of endotherms and hence is likely important for survival during energy bottlenecks, it also may enhance somatic growth when food is limited. We therefore hypothesize that torpor during the development in endotherms is far more widespread than is currently appreciated.     

Warm-up rates during arousal from torpor in heterothermic mammals: physiological correlates and a comparison with heterothermic insects

Summary This study examines the relationship between warm-up rate, body mass, metabolic rate, thermal conductance and normothermic body temperature in heterothermic mammals during arousal from torpor. Predictions based on the assumption that the energetic cost of arousal has been minimised are tested using data for 35 species. The observation that across-species warm-up rate correlates negatively with body mass is confirmed using a comparative technique which removes confounding effects due to the non-independence of species data due to shared common ancestry. Mean warm-up rate during arousal correlates negatively with basal metabolic rate and positively with the temperature difference through which the animal warms, having controlled for other factors. These results suggest that selection has operated to minimise the overall energetic, cost of warm-up. In contrast, peak warm-up rate during arousal correlates positively with peak metabolic rate during arousal, and negatively with thermal conductance, when body mass has been taken into account. These results suggest that peak warm-up rate is more sensitive to the fundamental processes of heat generation and loss. Although heterothermic marsupials have lower normothermic body temperatures and basal metabolic rates, marsupials and heterothermic eutherian mammals do not differ systematically in warm-up rate. Pre-flight warm-up rates in one group of endothermic insects, the bees, are significantly higher than predictions based on rates of arousal of a mammal of the same body mass.Abbreviations BMR basal metabolic rate - ICM independent comparisons method - MWR mean warm-up rate - PMR peak metabolic rate - PWR peak·warm-up rate - Tb_activity body temperature during activity - Tb_torpor body temperature during torpor - T _arousal increase in body temperature during arousal     

Effect of torpor on the water economy of an arid-zone marsupial,the stripe-faced dunnart (<Emphasis Type="Italic">Sminthopsis macroura</Emphasis>)

Metabolic rate and evaporative water loss (EWL) were measured for a small, arid-zone marsupial, the stripe-faced dunnart (Sminthopsis macroura), when normothermic and torpid. Metabolic rate increased linearly with decreasing ambient temperature (T_a) for normothermic dunnarts, and calculated metabolic water production (MWP) ranged from 0.85±0.05 (T_a=30°C) to 3.13±0.22 mg H₂O g^–1 h^–1 (T_a=11°C). Torpor at T_a=11 and 16°C reduced MWP to 24–36% of normothermic values. EWL increased with decreasing T_a, and ranged from 1.81±0.37 (T_a=30°C) to 5.26±0.86 mg H₂O g^–1 h^–1 (T_a=11°C). Torpor significantly reduced absolute EWL to 23.5–42.3% of normothermic values, resulting in absolute water savings of 50–55 mg H₂O h^–1. The relative water economy (EWL/MWP) of the dunnarts was unfavourable, remaining >1 at all T_a investigated, and did not improve with torpor. Thus torpor in stripe-faced dunnarts results in absolute, but not relative, water savings.     

Unusual nuclear structure of the spermatozoon in a marsupial,Sminthopsis crassicaudata

The organization of sperm chromatin in the dasyurid marsupial, Sminthopsis crassicaudata, was investigated using various morphological techniques. Transmission electron microscopy indicates two quite distinct chromatin regions became evident late in spermiogenesis with an outer globular region containing blocks of very electron-dense chromatin. Fluorescent light microscopical studies after staining with DNA dyes and 7-amino actinomycin D of testicular, caput, and cauda epididymal spermatozoa showed that this region fluoresced less brightly than the rest of the nucleus, indicating the presence of fewer DNA binding sites. Freeze fracture showed that the chromatin in most of the nucleus had randomly arranged particles of various sizes, but that of the outer region was composed entirely of small particles. This outer region was more resistant to low concentrations of the ionic detergent, SDS, whereas both guanidine hydrochloride and urea together with sodium chloride generally dispersed all the chromatin except that in the outer globular region and in a localized area of the nucleus beneath the acrosome. This study has thus revealed that the outer globular chromatin of these spermatozoa responds differently to ionic detergents and protein denaturing agents and has a different chromatin organization than most of the rest of the nucleus. The significance of these differences remains, however, to be determined. © 1994 Wiley-Liss, Inc.     

Ecology of natural hibernation in the marsupial mountain pygmy-possum (Burramys parvus)

The hibernating marsupial mountain pygmy-possum (Burramys parvus, 40 g) has to raise its slow-growing offspring during a short alpine summer. Only females provide parental care, while after mating males emigrate to marginal habitats often at lower altitudes which can sustain only low possum densities. We predicted that the hibernation strategies in mountain pygmy-possums are distinct from those of similar-sized placental hibernators, because of the developmental constraints in marsupials and because hibernation differs between the sexes. Using temperature-sensitive radio transmitters, we studied the hibernation patterns of free-living male and female mountain pygmy-possums living in a north- and a south-facing boulder field (Kosciusko National Park) for two consecutive winters. Individual possums commenced hibernation several months before the snow season. As in other hibernators, torpor in the mountain pygmy-possum was interrupted by periodic arousals which occurred most often during the late afternoon. Torpor bouts initially lasted a few days when the hibernacula temperature (T _hib) ranged from 4 to 7°C. As the hibernation season progressed, torpor bouts became longer and possum body temperatures (T _b) approached 2°C. The T _bs of females were significantly lower and torpor bouts were longer in the second half of the hibernation season than in males. Between torpor bouts, both sexes were often active and left hibernacula for periods of up to 5 days. Especially during the first months of the hibernation season, possums also frequently changed hibernacula sites probably in an attempt to select a site with a more suitable microclimate. Emergence from hibernation was closely coupled with the disappearance of snow from the possum habitat (September 1995, October 1996) and the limited fat stores probably dictate an opportunistic spring emergence. However, in 1995, spring was early and males emerged significantly earlier than females. In 1996, when snow melt was delayed, this difference vanished. Testes are regressed in males during hibernation and the time needed for testes growth and spermatogenesis favours an earlier emergence for males which was probably achieved by their preference for the more sun exposed north-facing boulder field. A sexual dimorphism in hibernation strategies and spring emergence therefore enables mountain pygmy-possums to cope with their harsh alpine environment. Received: 22 May 1997 / Accepted: 21 August 1997     

Ambient temperature cycles entrain the free-running circadian rhythms of the stripe-faced dunnart,Sminthopsis macroura

Summary We examined the effect of cycles of 12 h warm (35 ± 2 °C) and 12 h (21 ± 2 °C) ambient temperature (Ta) upon the circadian activity rhythms of stripe-faced dunnarts, Sminthopsis macroura, free-running in conditions of constant dark (DD) or constant light (LL). It was hypothesized that dunnarts would entrain to the temperature cycles (TaHLs) or show perturbations of period, and that LL would act synergistically with the TaHLs in these effects. Under DD, 2 of 6 animals showed clear entrainment to the TaHLs. Other animals exhibited changes of period () and heavy negative masking of activity during the warm fraction of the TaHLs. Under LL, 3 of 12 animals entrained to the TaHLs. It was concluded that Ta is a significant though weak Zeitgeber for S. macroura compared to light. It is possible that TaHLs entrain homeotherm activity rhythms by altering the rhythm of body temperature, which is usually tightly coupled to activity.Abbreviations TaHL a cycle of Higher and Lower ambient temperature - TaC Constant Ta - Tb body temperature     

Genetic variation in natural and laboratory populations of the marsupial Sminthopsis crassicaudata

Gel electrophoresis of blood proteins has detected allelic variation at five loci (TRF, PGD, SOD, ADA, GPI) in a laboratory colony of the dasyurid marsupial Sminthopsis crassicaudata. Family data show no significant departures from Mendelian expectations. Analysis of blood from wild-caught progenitors of the colony revealed significant differences in gene frequency between groups of animals captured from different parts of southern and central Australia and showed that there are two major population clusters. These interpopulation differences are particularly marked at the TRF locus and indicate that the river Murray is a barrier for this species.This project was supported by a grant from the Australian Research Grants Scheme.     

Identification of novel and known ovary-specific genes including ZP2, in a marsupial, the stripe-faced dunnart

During the early stages of oogenesis, oocyte-specific factors, synthesized by and stored within the oocyte, play critical roles during oogenesis, folliculogenesis, fertilization and early embryonic development in the mouse. The identification of marsupial maternal factors, expressed specifically in the ovary or oocyte, may provide an insight into the conserved evolutionary mechanisms that drive mammalian oocyte development to cleavage stages. In this study, 10 clones including dunnart ZP2 and c-mos, isolated by cDNA representational difference analysis, were validated by RT-PCR for ovary-specific expression. This novel combination of techniques to isolate ovary-specific genes has identified three novel genes with ovary-specific expression. Both dunnart ZP2 and c-mos exhibited ovary-specific expression, making this study the first isolation of c-mos in a marsupial species. Dunnart ZP2 expression was examined in detail by in situ hybridization and results indicate oocyte-specific expression of dunnart ZP2 in the cytoplasm of oocytes of primordial, primary and secondary follicles with expression being highest in oocytes of primary follicles. ZP2 was not expressed in granulosa cells of any follicles.     

The effect of metabolic fuel availability on thermoregulation and torpor in a marsupial hibernator

The physiological signal for torpor initiation appears to be related to fuel availability. Studies on metabolic fuel inhibition in placental heterotherms show that glucose deprivation via the inhibitor 2-deoxy-D-glucose (2DG) initiates a torpor-like state, whereas fatty acid deprivation via mercaptoacetate (MA) does not. As previous studies using inhibitors were limited to quantifying body temperature in placentals, we investigated whether inhibition of glucose or fatty acids for cellular oxidation induces torpor in the marsupial hibernator Cercartetus nanus, and how the response of metabolic rate is related to body temperature. Glucoprivation initiated a torpor-like state in C. nanus, but animals had much higher minimum body temperatures and metabolic rates than those of torpid food-deprived animals and arousal rates were slower. Moreover, 2DG-treated animals were thermoregulating at ambient temperatures of 20 and 12 °C, whereas food-deprived torpid animals were thermo-conforming. We suggest that glucoprivation reduces the hypothalamic body temperature set point, but only by about 8 °C rather than the approximately 28 °C during natural torpor. Reduced fatty acid availability via MA also induced a torpor-like state in some C. nanus, with physiological variables that did not differ from those of torpid food-deprived animals. We conclude that reduced glucose availability forms only part of the physiological trigger for torpor initiation in C. nanus. Reduced fatty acid availability, unlike for placental heterotherms, may be an important cue for torpor initiation in C. nanus, perhaps because marsupials lack functional brown adipose tissue.Abbreviations BAT brown adipose tissue - BMR basal metabolic rate - 2DG 2-deoxy-D-glucose - FD food deprived - GLM general linear models - MA mercaptoacetate - MR metabolic rate - RQ respiratory quotient - T_a ambient temperature - T_b body temperature - T_set body temperature set pointCommunicated by I.D. Hume     

The effect of temperature on adrenergic receptors of alveolar type II cells of a heterothermic marsupial

Fat-tailed dunnarts, Sminthopsis crassicaudata, survive dramatic changes in body temperature during torpor without experiencing surfactant dysfunction. Adrenergic factors regulate surfactant secretion through beta(2)-adrenergic receptors on alveolar type II cells. Temperature has no effect on the secretory response of dunnart type II cells to adrenergic stimulation. We hypothesise that during torpor, dunnart type II cells up-regulate the number of adrenergic receptors present on type II cells to enable stimulation at lower concentrations of agonist. Here, we isolated type II cells from warm-active (35 degrees C) and torpid (15 degrees C) dunnarts and examined the effects of an in vitro temperature change on the number and activity of adrenergic receptors. Torpor did not affect the beta-adrenergic receptor number. However, we observed a significant decrease in adrenergic receptor number when cells from warm-active animals were incubated at 15 degrees C and when cells from torpid animals were incubated at 37 degrees C. cAMP production was significantly higher in type II cells from torpid dunnarts than warm-active dunnarts and this may contribute, in part, to the temperature insensitivity we have previously observed in the adrenergic regulation of surfactant secretion.     

Desmoglein‐2 during pregnancy and its role in the evolution of viviparity in a marsupial (Sminthopsis crassicaudata; Dasyuridae)

Attachment of the blastocyst and formation of the placenta during pregnancy is dependent on structural and cellular changes occurring in the uterine epithelium and in particular to the plasma membrane of these uterine cells. Desmosome expression decreases during pregnancy in eutherians and some squamates, presumably allowing for remodeling of the uterine epithelium and invasion of the trophoblast during implantation. Marsupials are a distinct mammalian amniote lineage of viviparity, with a short implantation or attachment period and varying levels of invasive placentation. To test the generality of changes to the uterine epithelium during pregnancy across mammals, we characterized the distribution of desmosomes in the uterine epithelial cells of a marsupial, Sminthopsis crassicaudata, using electron microscopy and immunohistochemistry. The absolute number of desmosomes along the lateral plasma membrane decreases during pregnancy and desmosomes are redistributed towards the apical region of the lateral plasma membrane as pregnancy proceeds, similar to what occurs during pregnancy in eutherian mammals. Despite the lower level of maternal investment in pregnancy and the noninvasive structure of fetal membranes in marsupials there are similarities in number and redistribution of desmosomes along the plasma membrane and changes to the morphology of the uterine epithelial cells suggesting that similar plasma membrane changes occur across all lineages of amniote vertebrates. J. Morphol. 276:261–272, 2015. © 2014 Wiley Periodicals, Inc.     

Foraging strategies of an insectivorous marsupial,Sminthopsis youngsoni (Marsupialia: Dasyuridae), in Australian sandridge desert

This study investigated the effects of predation risk, dune position and microhabitat on foraging of the Lesser Hairy‐footed Dunnart Sminthopsis youngsoni, a small insectivorous marsupial, in the Simpson Desert of western Queensland. The intensity of foraging was assessed by establishing feeding stations (dishes containing mealworms) in open and bush microhabitats at three levels on sand dunes, and recording the numbers of prey taken by dunnarts from the stations after nightly bouts of foraging. Risk of predation was manipulated by provision of artificial illumination at alternate feeding stations on each of five occasions when trials were run. The numbers of mealworms left after feeding bouts varied inconsistently across trials, providing little evidence that dunnarts respond to habitat or predation risk while foraging. These results contrast sharply with studies of arid zone rodents, where foraging is usually sensitive to both predation risk and resource distribution. We suggest that S. youngsoni forages equally in all habitats of its sandridge environment, and experiences relatively low risk of predation whilst doing so.