Biochemical studies of intrauterine components of the tammar wallaby Macropus eugenii during pregnancy

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 325-338
Author(s):  
Elizabeth J. Thornber ◽  
Marilyn B. Renfree ◽  
Gregory I. Wallace
Keyword(s):  
Protein Concentration ◽  
Specific Binding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Fold Increase ◽  
Tammar Wallaby ◽  
Macropus Eugenii ◽  
Tenfold Increase ◽  
Specific Proteins ◽  
Wet Weight

The in vitro uptake and incorporation of [3H]ui idine by blastocysts of the tammar wallaby showed a 16- and 30-fold increase from day 0 to day 10 after removal of pouch young, respectively. Two of the six non-expanded blastocysts recovered on day 5 showed a tenfold increase in incorporation. During the first ten days after removal of pouch young the diameter of the blastocyst increased threefold. Endometrial exudate from gravid uteri had a higher protein concentration than exudate from nongravid uteri (39·5 ± 0·9 and 32·0 ± 2·0 mg/ml (mean ± s.e.m.), respectively). Endometrial exudates from uteri where the blastocyst was actively growing were found to contain six uterine-specific proteins. These were separated by gradient polyacrylamide gel electrophoresis. Two of the proteins were pre-albumins and the others were larger molecules (M.W. 153000–670000). Two proteins were only present at particular stages of pregnancy: the other four were present at all stages from diapause to birth, in exudate from gravid and nongravid uteri. The specific binding of progesterone and androstenedione to proteins in endometrial exudates or uterine flushings from pregnant wallabies was less than one per cent of the value obtained from day-5 pregnant rabbits. The ability of mouse blastocysts to take up and incorporate [3H]uridine into acidinsoluble material increased threefold in the presence of day-10 endometrial exudates from wallabies. However, this was less than ten percent of the values obtained in the presence of bovine serum albumin. The concentration of calcium in endometrial exudates increased from 23·6 to 45·2 μg/ml during pregnancy; in endometrium it remained at 88·7 μg/g (wet weight) throughout pregnancy, and in plasma it was 53·3 μg/ml. The concentration of zinc in endometrial exudates was 4·5 μg/ml; in endometrium it decreased from 21·8 to 13·3 μg/g (wet weight) during pregnancy and in plasma it was 0·6 μg/ml.

PROLACTIN RECEPTORS IN THE MAMMARY GLAND, CORPUS LUTEUM AND OTHER TISSUES OF THE TAMMAR WALLABY, MACROPUS EUGENII

1979 ◽  
Vol 83 (1) ◽  
pp. 79-89 ◽  
Author(s):  
C. SERNIA ◽  
C. H. TYNDALE-BISCOE
Keyword(s):  
Mammary Gland ◽  
Corpus Luteum ◽  
Specific Binding ◽  
Tammar Wallaby ◽  
Binding Activity ◽  
Breeding Cycle ◽  
Macropus Eugenii ◽  
Lactating Mammary Gland ◽  
The Mean ◽  
Ovine Prolactin

SUMMARY Specific binding of radio-iodinated ovine prolactin to subcellular tissue fractions of the tammar wallaby (Macropus eugenii) was investigated. Specific binding was found, in order of decreasing binding activity, in the lactating mammary gland, corpus luteum, corpus albicans, adrenal gland and ovary. Specific binding was absent in kidney, liver, brain and inactive mammary gland. The mean association constant (Ka at 23 °C) was determined as 0·90 × 109, 2·20 × 109, 2·44 × 109, 3·38 × 109 and 10·98 × 1091/mol for mammary gland, adrenal, corpus albicans, corpus luteum and ovary respectively. The mean receptor concentration (N) varied from 92·87 × 10−14 mol/mg protein for the mammary gland to 1·03 × 10−14 mol/mg protein for the ovary. The concentration in the corpus luteum varied between tissue pools collected at different times of the annual breeding cycle. The specificity for prolactin was shown in the mammary gland and corpus luteum by the failure of ovine FSH, LH, GH and TSH to displace 125I-labelled ovine prolactin, whereas it was displaced readily by both ovine and bovine prolactin.

Plasma protein polymorphisms in the tammar wallaby, Macropus eugenii

1998 ◽  
Vol 46 (2) ◽  
pp. 193 ◽  
Author(s):  
H. Arthur ◽  
K. Bell ◽  
D. W. Cooper
Keyword(s):  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
South Australia ◽  
Backcross Progeny ◽  
Tammar Wallaby ◽  
Vitamin D Binding Protein ◽  
Macropus Eugenii ◽  
Human Proteins ◽  
Protein Polymorphisms

Five populations of the Australian tammar wallaby, Macropus eugenii, from Kangaroo Island, South Australia, and Garden, Abrolhos and Middle Islands and Perup, Western Australia, were examined for plasma protein polymorphisms. Select Kangaroo/Garden Island hybrids and backcross progeny were also included in the study. Vitamin D binding protein (GC), albumin (ALB), transferrin (TF), protease inhibitor (PI), haemopexin (HX), haptoglobin (HP) and immunoglobulin G (IgG) were identified by polyacrylamide gel electrophoresis, pH 7.9, isoelectric focusing, pH 4.2–4.9, and immunoblotting with rabbit antisera to human proteins. Five GC (A, B, C, D, E), two ALB (A, B), two TF (A, B) and five PI (I, J, L, M, P) variants were detected, and limited family studies demonstrated a codominant allelic inheritance for each of the systems.

Pellet-freezing spermatozoa of two marsupials: the tammar wallaby, Macropus eugenii, and the brushtail possum, Trichosurus vulpecula

1996 ◽  
Vol 8 (4) ◽  
pp. 681 ◽  
Author(s):  
FC Molinia ◽  
JC Rodger
Keyword(s):  
Room Temperature ◽  
Tammar Wallaby ◽  
Brushtail Possum ◽  
Vitro Fertilization ◽  
Macropus Eugenii ◽  
Progressive Motility ◽  
Viable Population ◽  
Phosphate Buffered Saline ◽  
Breeding Techniques

A protocol was developed for pellet-freezing spermatozoa of the tammar wallaby and the brushtail possum. Seren was collected by electro-ejaculation and wallaby spermatozoa were washed by 'swim-up' into phosphate-buffered saline (PBS), whereas possum spermatozoa were not washed. Wallaby spermatozoa were screened for toxicity in diluents containing a range of cryoprotectants (0-10%): dimethyl sulfoxide (DMSO), ethylene glycol and propanediol. Possum spermatozoa were tolerant of diluents containing 17.5% glycerol. Wallaby and possum spermatozoa were diluted 1:1 with the most promising cryoprotective diluents (final concentrations in PBS: possum, 17.5% glycerol; wallaby, 7.5% glycerol + 10% DMSO) and, after 5 min equilibration at room temperature, were pellet-frozen. Pellets were thawed (35 degrees C) and wallaby spermatozoa were washed by centrifugation (200 g for 5 min) and resuspended in PBS to minimize cryoprotectant toxicity. A high proportion of possum spermatozoa was recovered after freezing (67.5%), having good progressive motility (3.6 on a 0-5 scale). The progressive motility of frozen-thawed wallaby spermatozoa was also high (3.0), but only 10% of motile spermatozoa were recovered. The pellet-freezing method in conjunction with the post-thaw washing procedure (wallaby) may produce a viable population of cryopreserved marsupial spermatozoa suitable for use in assisted-breeding techniques such as in vitro fertilization and artificial insemination.

scholarly journals Induction of sperm maturation in vitro in epididymal cell cultures of the tammar wallaby (Macropus eugenii): disruption of motility initiation and sperm morphogenesis by inhibition of actin polymerization

Reproduction ◽  
2002 ◽  
pp. 107-117 ◽  
Author(s):  
M Lin ◽  
R Hess ◽  
RJ Aitken
Keyword(s):  
Culture System ◽  
Actin Polymerization ◽  
Tammar Wallaby ◽  
Sperm Maturation ◽  
Actin Organization ◽  
Cell Monolayers ◽  
Macropus Eugenii ◽  
Maturation In Vitro ◽  
Marsupial Species

A sperm-epididymal cell co-culture was shown to be capable of inducing the in vitro maturation of spermatozoa from a marsupial species, the tammar wallaby (Macropus eugenii). This system was able to maintain wallaby epididymal epithelial cells in vitro for more than 2 months. The system also enabled immature wallaby spermatozoa to differentiate from a T-shaped to a streamlined form, accompanied by the development of progressive motility after co-culture with epididymal cell monolayers that had been cultured for 7 days. The addition of inhibitors of actin polymerization (latrunculin A or B) to the co-culture system showed that wallaby sperm maturation was impaired by the interruption of actin organization within the immature spermatozoa. These results indicate that actin filaments play a significant role in sperm transformation during post-testicular maturation in marsupials. These observations also indicate that the marsupial co-culture system has the potential to greatly increase understanding of sperm-epididymal cell interactions and the mechanism of sperm maturation in these species.

Fibroblasts of rabbit kidney in culture. I. Characterization and identification of cell-specific markers

1991 ◽  
Vol 261 (2) ◽  
pp. F283-F291 ◽  
Author(s):  
H. P. Rodemann ◽  
G. A. Muller ◽  
A. Knecht ◽  
J. T. Norman ◽  
L. G. Fine
Keyword(s):  
Renal Cortex ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cell Lineage ◽  
Rabbit Kidney ◽  
Protein Markers ◽  
Postmitotic Cells ◽  
Specific Proteins ◽  
Population Doublings ◽  
Two Populations

There is currently no information available as to whether different renal fibroblast subpopulation can be identified and whether they show differences in functional properties. We therefore compared the growth characteristics of interstitial fibroblasts derived from the rabbit renal cortex and inner medulla (papilla) and sought cell-specific markers for the two populations of cells. Analyses of the population dynamics revealed that the mitotic lifespan of papillary fibroblasts (PF) is approximately 50% longer than that of cortical fibroblasts (CF), with the former going through 20 cumulative population doublings (CPD) before transition into terminally differentiated postmitotic cells compared with 9 CPD in CF. PF and CF populations contained three types of mitotically active cells (MFI, MFII, MFIII) and three types of postmitotic cells (PMFIV, PMFV, PMFVI) differentiating along a terminal cell lineage from MFI through PMFVI. In both PF and CF cultures the percent of MF-type cells decreased and the percent of postmitotic cells increased with successive doublings. Two-dimensional polyacrylamide gel electrophoresis of uniform clonal populations of MFIII-type cells revealed two specific proteins for PF-MFIII-type cells, pf1 and pf2, and three specific proteins for CF-MFIII-type cells, cf1, cf2, and cf3. Additionally, a monoclonal antibody was raised that does not recognize CF in culture, but reacts strongly with PF. These studies demonstrate that rabbit renal PF have a pattern of growth in vitro that is distinct from that of CF and that they can be positively identified by specific immunological and protein markers in vitro.

scholarly journals In-vitro secretion of progesterone by the corpus luteum of the tammar wallaby, Macropus eugenii

Reproduction ◽  
1983 ◽  
Vol 67 (1) ◽  
pp. 57-63 ◽  
Author(s):  
L. A. Hinds ◽  
S. M. Evans ◽  
C. H. Tyndale-Biscoe
Keyword(s):  
Corpus Luteum ◽  
Tammar Wallaby ◽  
Macropus Eugenii

scholarly journals Changes in the Milk Proteins during Lactation in the Tammar Wallaby, Macropus eugenii

1982 ◽  
Vol 35 (2) ◽  
pp. 145 ◽  
Author(s):  
Stuart W Green ◽  
Marilyn B Renfree
Keyword(s):  
Protein Concentration ◽  
Post Partum ◽  
Whey Proteins ◽  
Milk Proteins ◽  
Tammar Wallaby ◽  
Total Protein Concentration ◽  
Mean Values ◽  
Macropus Eugenii ◽  
Stage Of Lactation ◽  
Whole Milk

Samples of whey proteins from the milk of tammar wallabies, Macropus eugenii, were examined by acrylamide gel electrophoresis at all stages of lactation up to 280 days post partum. Whey albumin, ,B-globulin and y-globulin fractions had similar electrophoretic mobility to that of the equivalent serum protein fractions, but the proteins in the IX-globulin and pre-albumin regions differed markedly. The IX-globulins are presumed polymorphic because individuals at the same stage of lactation showed great variability in these electrophoretic regions: up to five polymorphic bands were recognized. Milk proteins changed qualitatively throughout lactation, and in particular the concentration of the pre-albumin and IX-globulin fractions increased from approximately day 180 to the end of lactation. Total protein concentration of both whole milk and whey approximately doubled in the second half of lactation compared to the first half, reaching maximum mean values of 114 � 47 and 96 � 50 g 1- 1 , respectively. Whole milk contained consistently more protein than whey, presumably due to the casein it contains.

239. Expression of PAF-R and p53 in the endometrium during entry into and reactivation from diapause in the tammar wallaby

2008 ◽  
Vol 20 (9) ◽  
pp. 39
Author(s):  
J. C. Fenelon ◽  
G. Shaw ◽  
M. B. Renfree
Keyword(s):  
Tammar Wallaby ◽  
Early Embryo ◽  
Human Reproduction ◽  
Embryonic Diapause ◽  
P53 Expression ◽  
Cellular Location ◽  
Macropus Eugenii ◽  
A Cell

Embryonic diapause is widespread amongst mammals, but is especially common in the kangaroos and wallabies. In the tammar, Macropus eugenii, the sequence of endocrine events leading to embryonic diapause and reactivation are well defined and the blastocyst can remain in diapause for up to 11 months without cell division or apoptosis occurring (Renfree and Shaw 2000). The ovarian hormones exert their effects on the blastocyst by alterations in the endometrial secretions, but the molecular cross-talk between the endometrium and blastocyst is unknown. One possible regulator of diapause is the phospholipid PAF, an embryotrophin that acts as a trophic/survival factor for the early embryo (O'Neill 2005) partly by inactivating the expression of p53, a cell cycle inhibitor, via the PI3-K pathway. PAF is released from the tammar endometrium around the time of reactivation from diapause (Kojima et al. 1993). This study examined the expression of PAF-R and p53 in the tammar endometrium at entry into, and reactivation from, diapause. PAF-R and p53 were highly conserved with orthologueues in human and mouse. PAF-R and p53 expression was assessed by RT–PCR and both genes were expressed in the endometrium at all stages examined. Quantitative PCR (QPCR) studies performed for PAF-R in the endometrium show that levels of PAF-R vary depending on the stage examined and appear to be increasing at entry into diapause and decreasing at exit from diapause. Immunohistochemical (IHC) studies are in progress to determine the cellular location of PAF-R in the endometrium and confirm the QPCR results. QPCR and IHC studies are in progress to determine if there is any change in levels of expression or cellular location of p53 between the stages examined and how this relates to PAF-R availability. These results suggest that the control of diapause in the tammar involves interactions between multiple factors. (1) Renfree MB, Shaw G (2000) Diapause. Annu Rev Physiol 62, 353–375 (2) O'Neill C (2005) The role of Paf in embryo physiology. Human Reproduction Update 11, 215–228 (3) Kojima T et. al. (1993) Production and secretion of progesterone in vitro and presence of PAF in early pregnancy of the marsupial, Macropus eugenii. Reproduction Fertility Development 5, 15–25.

scholarly journals The Two-Component System PhoPR of Clostridium acetobutylicum Is Involved in Phosphate-Dependent Gene Regulation

2008 ◽  
Vol 190 (20) ◽  
pp. 6559-6567 ◽  
Author(s):  
Tomas Fiedler ◽  
Maren Mix ◽  
Uta Meyer ◽  
Stefan Mikkat ◽  
Michael O. Glocker ◽  
...  
Keyword(s):  
Clostridium Acetobutylicum ◽  
Response Regulator ◽  
Specific Binding ◽  
Expression Patterns ◽  
Regulatory System ◽  
Fold Increase ◽  
Growth Conditions ◽  
Two Dimensional Gel Electrophoresis ◽  
Two Component

ABSTRACT The phoPR gene locus of Clostridium acetobutylicum ATCC 824 comprises two genes, phoP and phoR. Deduced proteins are predicted to represent a response regulator and sensor kinase of a phosphate-dependent two-component regulatory system. We analyzed the expression patterns of phoPR in Pi-limited chemostat cultures and in response to Pi pulses. A basic transcription level under high-phosphate conditions was shown, and a significant increase in mRNA transcript levels was found when external Pi concentrations dropped below 0.3 mM. In two-dimensional gel electrophoresis experiments, a 2.5-fold increase in PhoP was observed under Pi-limiting growth conditions compared to growth with an excess of Pi. At least three different transcription start points for phoP were determined by primer extension analyses. Proteins PhoP and an N-terminally truncated *PhoR were individually expressed heterologously in Escherichia coli and purified. Autophosphorylation of *PhoR and phosphorylation of PhoP were shown in vitro. Electromobility shift assays proved that there was a specific binding of PhoP to the promoter region of the phosphate-regulated pst operon of C. acetobutylicum.

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