Oestrous cycle of captive southern hairy-nosed wombats (Lasiorhinus latifrons) in South Australia, Australia

There is limited information available on the oestrous cycle of female southern hairy-nosed wombats (Lasiorhinus latifrons). This is mainly due to an extremely poor breeding success in captivity and the difficulty in routine recapturing of these cryptic, semi-fossorial animals in the wild. The aim of this study was to characterise the oestrous cycle of this species by monitoring peripheral plasma concentrations of progesterone and oestradiol, assessing changes in vaginal cytology, pouch condition and the urogenital sinus. Eight adult female wombats were monitored during the breeding season (July-December) over 2 years (2002-2003). Samples were collected up to three times a week. Vaginal smears contained several cell types, categorised by morphology, as either superficial epithelial cells or parabasal-intermediate cells. Leucocytes were also counted. Plasma progesterone profiles showed a mean oestrous cycle length of 36.33+/-0.67 days with a peak progesterone concentration of 139.53+/-10.62nmol/L. Levels of oestradiol peaked at a mean level of 467.33+/-44.32pmol/L on average 5 days before a rise in plasma progesterone values. The proportion of epithelial cells in vaginal smears varied throughout the cycle, with a high percentage of superficial epithelial cells observed during the follicular phase. During periods when progesterone concentrations were high, a greater percentage of parabasal-intermediate cells was observed. In conclusion, this study has characterised the oestrous cycle of the southern hairy-nosed wombat and confirmed that changes in vaginal smears together with pouch and urogenital sinus details could be used to determine signs of oestrus in this species.     

Social influences on oestrous cycle length and plasma progesterone concentrations in the female lesser mouse lemur (Microcebus murinus)

Plasma progesterone concentrations were recorded during one breeding season in 19 lesser mouse lemur females living in different social conditions. The oestrous cycle length and the progesterone profile mainly depended on the social environment of the female. For totally isolated females, the oestrous cycle lasted 38 +/- 5.7 days and included a 25-30-days spontaneous luteal phase with a progesterone peak about 100 ng/ml between the 20th and 25th days after oestrus, and a prolonged preovulatory period of 10-15 days which could be considered equivalent to the follicular phase of a menstrual cycle. When females were able to communicate through olfactory, visual and auditory signals, the oestrous cycle was significantly lengthened (53.7 +/- 5.9 days). When females had tactile contacts, the oestrous cycle was further lengthened (62.7 +/- 0.8 days). This lengthening of the oestrous cycle was related to an extension of the luteal phase associated with a decrease in progesterone concentrations during this period. In females maintained with one male (paired) or with males and females (heterosexually grouped), large individual variations were shown in cycle lengths or in progesterone concentrations. In these females, cycle lengths and progesterone concentrations were inversely correlated to plasma cortisol concentrations.     

Faecal progesterone metabolites and behavioural observations for the non-invasive assessment of oestrous cycles in the common wombat (Vombatus ursinus) and the southern hairy-nosed wombat (Lasiorhinus latifrons)

Wombats belong to Australia's unique marsupial species. Two of the three remaining species, the common wombat (Vombatus ursinus) and the southern hairy-nosed wombat (Lasiorhinus latifrons) are abundant. The third species, the northern hairy-nosed wombat (Lasiorhinus krefftii) has only about 115 individuals left in the wild. This study aimed to gain further insight into the basic reproductive biology of wombat species and evaluate the value of faecal progesterone metabolites and behavioural patterns as a means for non-invasive monitoring of the oestrous cycle in common and the southern hairy-nosed wombats. In an initial study, three different faecal steroid assays showed that 20alpha-OH-pregnanes were the main progesterone metabolites. These metabolites were examined in captive female common wombats (n = 5) and southern hairy-nosed wombats (n = 2). In one female common wombat 11.7 days with a follicular phase of 25.6 +/- 6.3 days and a luteal phase of 28.2 +/- 12.7 days. The data for faecal pregnanes obtained in the southern and in one male common wombat oestrous related behavioural data were obtained. Individual cycling females exhibited a significant relationship between plasma progesterone and faecal pregnanes. In the common wombat, the values for faecal pregnanes showed an oestrous cycle length of 55.1 +/- hairy-nosed wombat during the breeding season gave an oestrous cycle length of 41.1 +/- 12.8 days with a follicular phase of 27.9 +/- 12.3 days and a short luteal phase of 13.3 +/- 1.1 days. The behavioural data show that the faecal sniffing behaviour of the male, tended to increase around the time that oestrous was found. In conclusion, monitoring of 20alpha-OH-pregnanes in wombat faeces could be a useful methodology to monitor reproductive cycles in the wombat, and can possibly be applied to monitor the endangered northern hairy-nosed wombat.     

Plasma,milk and faecal progesterone concentrations during the oestrous cycle of lactating dairy cows with different milk yields

The hypotheses tested in this study were that neither average progesterone (P4) concentrations in plasma and milk nor average progesterone metabolites concentrations in faeces would differ during an oestrous cycle in two groups of cows with differing daily milk yields. High producing (HP = 8) and low producing (LP = 8) dairy cows were selected randomly for the study. Their oestrous cycles were initially synchronised using P4 and prostaglandin F2alpha. Chromic oxide capsules were administered twice daily to measure total faecal output. Samples of blood, faeces and milk were taken daily throughout one oestrous cycle, plasma and milk P4, and faecal progesterone metabolites (FP4M) assayed. The average daily milk yields in the two groups were 30.8 and 21.9l per day, respectively (P < 0.0001), although daily faecal output was similar in both the groups (HP, 7.7 versus LP, 6.9 kg DM; P = 0.24). Mean plasma and milk P4 concentrations were similar in both the groups (plasma P4, 4.12 versus 4.05 ng/ml; P = 0.3; milk P4, 8.2 versus 8.3; P = 0.9) during dioestrus. Average daily excretion of P4 to the milk was greater in HP than LP cows (252 versus 185 microg, P = 0.04). Neither concentration nor the daily yield of FP4Ms was affected by level of milk yield (concentration: 12.2 versus 11.5 microg/g; daily yield: 89.1 versus 82.9 mg per day; P > 0.05). These data showed that the concentrations of P4 in plasma and milk, and the concentrations and daily yields of FP4M were not affected by the level of daily milk yields which differed by about 41% of the LP average of 21.9l.     

Vaginal oestrus during the reproductive and non-reproductive period in European ground squirrels

Most hibernating animals are thought to be monoestrous because reproductive activity is subject to strong time constraints. In previous studies, female European ground squirrels (Spermophilus citellus) turned out to have elevated oestradiol levels during late lactation and after weaning, indicating ovarian activity during summer. Therefore, we monitored vaginal cytology, endocrine changes, and vulval development in semi-free-living female European ground squirrels throughout one active season. Vaginal oestrus, defined by the predominance of cornified cells in smear samples, was found during the mating period shortly after vernal emergence. This phase was followed by metoestrus, characterised by the appearance of leukocytes, and a subsequent anoestrous phase. During weaning or postlactation, a second vaginal oestrus was documented in all experimental females, again followed by a metoestrous and an anoestrous phase lasting until hibernation. In line with the second vaginal oestrus, plasma oestradiol concentrations peaked during postlactation. Progesterone levels were elevated from gestation to postlactation, and titres were marginally higher during vaginal oestrus in summer than in spring. Vulval swelling was more pronounced during the first than the second vaginal oestrus. The second oestrous cycle was non-reproductive, as males were sexually inactive with regressed testes during summer. We assume that the second oestrous cycle and the accompanying endocrine changes have beneficial effects on prehibernatory fattening and reproductive performance in the subsequent season. This might allow females to become oestrous immediately after emergence from hibernation in spring.     

Plasma concentrations of progesterone and testosterone in captive woolly opossums (Caluromys philander)

Plasma testosterone and progesterone concentrations were measured in captive woolly opossums, a didelphid marsupial originating from neotropical forests in French Guyana. Although not exposed to cyclic environmental conditions as in the field, both sexes exhibited spontaneous circannual changes in sexual hormones. Males showed synchronous variations in plasma testosterone characterized by significant elevated concentrations during April and September (8.6 +/- 1 ng/ml, N = 5) and lower levels from May to July (3.6 +/- 0.4 ng/ml). In females, synchronous periods of 2-3 successive oestrous cycles occurred. Between these periods, females remained acyclic. The oestrous cycle, determined by urogenital smears, lasted 28-45 days (n = 14) and included a 20-day spontaneous luteal phase in which progesterone concentrations reached 30-40 ng/ml plasma. Even though testosterone concentrations in paired males increased significantly in response to oestrous periods of the paired females, spontaneous circannual rhythms of sexual activity were not well synchronized between the sexes in captivity. When compared to field data, sexual activity of captive animals followed a pattern similar to that in wild animals, without any changes in males but with a delay of 3 months in females.     

Progesterone and testosterone concentrations during oestrous cycle and pregnancy in the common vole (Microtus arvalis Pallas)

Serum progesterone and testosterone concentrations were measured during different stages of oestrous and pregnancy in paired and unpaired female common voles (Microtus arvalis). Hormone concentrations were measured by ELISA, and cycle stages were determined by vaginal smears. Paired females usually had serum progesterone concentrations of more than 10 ng/ml in the oestrous cycle. A significant maximum was detected in prooestrous (51.70 +/- 7.84 ng/ml, mean +/- S.D.). Serum progesterone concentrations increased from about 40 ng/ml at the beginning of pregnancy to about 70 ng/ml on days 15 and 16. The last 2 days before parturition (days 19 and 20) were characterised by a decrease of progesterone concentrations to ca. 30 ng/ml. The maximum concentration of testosterone was found in prooestrous (1.58 +/- 0.31 ng/ml). Concentrations during pregnancy varied between 1.5 and 2.1 ng/ml. In two of three cases unpaired females exhibited progesterone values below 10 ng/ml, but with varying vaginal smear patterns. The combination of progesterone concentrations and vaginal smear patterns was found to be regular in only 23.8% of the cases. The most frequent cycle stage found was the oestrous (44.2%). Mean concentrations of progesterone (10.43 +/- 13.81 ng/ml) and testosterone (0.85 +/- 1.11 ng/ml) in unpaired females were significantly lower than in paired females, thereby denoting reproductive inactivity in the former. The study presents basic data for several parameters of the reproductive biology in the common vole and confirms the importance of combining hormone assays and vaginal smear monitoring in reproductive research.     

Plasma progesterone concentrations and length of the first spontaneous oestrous cycle in pubertal rats

The length of the first spontaneous oestrous cycle in pubertal Wistar-Imamichi strain rats determined by vaginal smears varied from 5 to 18 days. The variation was ascribed to the period (3-16 days) of the stage of vaginal smears consisting of leucocytic cells (L stage). Plasma progesterone concentration and the decidual reaction in the uterus were used as indicators of the function of the corpus luteum and the L stage period was categorized as short, lasting for 3-6 days (average 4 days) with non-functional corpora lutea, or long, lasting 9-16 days (average 12 days) and with functional corpora lutea. Rats with the long L stage showed nocturnal and diurnal prolactin surges, but no daily changes in prolactin values were observed in rats with a short L stage. Daily changes in prolactin concentrations were maintained by the administration of progesterone in rats ovariectomized on Day 6 of the L stage. Plasma progesterone values on Day 6 of the L stage decreased with ergocornine treatment on Days 4 and 5 of the L stage and administration of bovine prolactin restored the level. These results indicate that the L stage observed in the first oestrous cycle is maintained by a positive feedback relation between progesterone and prolactin secretions.     

Changes in electrical impedance of the vaginal medium during the menstrual cycle of female rhesus monkeys (Macaca mulatta)

Changes in electrical impedance of the vaginal medium during the menstrual cycle were recorded in female Rhesus monkeys using electrical probe and were correlated with estradiol-17 beta and progesterone plasma concentrations. A gradual decrease in impedance was observed during the follicular phase, the lowest values being observed between days 12-17 of the cycle. Impedance increased again during the first third of the luteal phase until day 21. The reversal of the impedance gradient's sign was nearly concomitant with the appearance of a detectable plasma progesterone concentration. These results support the use of vaginal impedance measurements as a help for the diagnosis of the periovulatory time in the female Rhesus monkey.     

Characteristics of the oestrous cycle of Iberian red deer (Cervus elaphus hispanicus) assessed by progesterone profiles

This study examines the length of the oestrous cycle in 16 Iberian red deer females assessed by means of changes in progesterone concentrations, along with the changes in the profile of this hormone. Samples were collected three occasions per week from the week after calving (15 May to 15 June) up to May of the following year. The oestrous cycle lasted 19.57+/-0.29 days (range 10-27 d) calculated in 130 oestrous cycles examined. Progesterone titres did not rise above 0.5 ng/ml in the follicular phase, except in four samples. The maximum peak in progesterone concentration during the luteal phase remained above 1 ng/ml in most cases. Twenty-five percent of the individuals studied (4 out of 16) showed an oestrous cycle lasting shorter than the mean (15.2+/-0.30 days) before the start of the reproductive season, followed by a period of sexual inactivity. The standard progesterone profile in natural oestrous cycles rose from basal levels to those above 0.5 ng/ml four days after onset of oestrus, reached a peak of 1.71+/-0.07 ng/ml and then declined to less than 0.2 ng/ml after day 20. Following the rapid decline of progesterone after day 14, the concentration remained around the baseline level of 0.1 to 0.2 ng/ml during the immediate pre- and post-ovulatory phase of the cycle.     

Circulating progesterone and oestradiol-17 beta concentrations in cyclic Cape porcupines, Hystrix africaeaustralis

The general pattern of steroid secretion during the 30-day oestrous cycle of the Cape porcupine is that of a surge (25-176 pg/ml) in oestradiol-17 beta secretion at the time of perforation of the vaginal closure membrane, followed by an increase in progesterone concentrations, the latter attaining peak values (mean 5.9 +/- 2.1 ng/ml) 8-19 days (13.8 +/- 2.8 days) after vaginal opening. Copulation occurred after the oestradiol-17 beta surge and the length of the luteal phase of the cycle varied from 21 to 35 days (29.3 +/- 4.7 days), this representing 93% of the length of the cycle. Perforation of the vaginal closure membrane was not always accompanied by an increase in oestradiol-17 beta levels and some instances (19%) of vaginal opening were not followed by an increase in progesterone secretion. The hormonal characteristics of the oestrous cycle of females housed with vasectomized males were similar to those of females housed with intact males.     

Plasma concentrations of progesterone, oestrogens, testosterone and LH-like activity during the oestrous cycle of the camel (Camelus dromedarius)

Peripheral plasma concentrations of progesterone, total oestrogens and testosterone (measured by RIAs) and LH (monitored by the mouse Leydig cell bio-assay) were measured in 8 female camels for a complete oestrous cycle (23.1 +/- 1.2 days). The absence of an LH surge and a low concentration of progesterone (less than 1 ng/ml) during oestrus (5 days) and throughout the cycle indicated a failure of spontaneous ovulation and absence of a subsequent luteal phase in this species. High concentrations of testosterone and oestrogens indicated that the oestrous cycle in the camel is mostly follicular and that the increasing values of the two hormones during follicular development (5 days) is probably the stimulus to behavioural oestrus.     

The effects of stress and of ACTH administration in hormone profiles, oestrus and ovulation in pigs

Hormone concentrations and oestrous cycle patterns were studied in five chronically cannulated gilts. During oestrous cycles that were unaffected by stress, plasma oestrogen concentrations remained at basal luteal phase levels (10 to 30 pg/ml) until plasma progesterone had decreased to less than 2 ng/ml. The pre-oestrus surge of oestrogen ranged from 40 to 80 pg/ml. Plasma corticoid concentrations varied randomly and were not related to oestrogen, progesterone concentrations, or the stage of the oestrous cycle. There was, however, evidence of a positive relationship between elevated corticoid levels and observed stressful events. The stress of surgery or illness acting during the follicular phase of the oestrous cycle delayed the onset of oestrus, and corticoid levels were frequently elevated on these occasions. Elevated plasma corticoid concentrations in response to ACTH treatment were associated with either a change in the timing of or a suppression of the pre-oestrus LH peak. Altering the timing of the LH peak resulted in the formation of large partially luteinized ovarian cysts, while suppressing LH interfered with follicular development and led to small ovarian cysts. These experiments suggest that stress acting via the adrenal gland may play a role in the aetiology of infertility in sows.     

Concentrations of progesterone in the backfat of pigs during the oestrous cycle and after ovariectomy

Small samples of backfat were taken daily during one oestrous cycle and more frequently after ovariectomy from 12 gilts by means of a simple biopsy technique and the levels of progesterone were determined. Compared to the levels of progesterone in peripheral plasma changes in backfat levels during the oestrous cycle were delayed by 1-2 days. Maximal levels with 89.7 +/- 9.2 (mean +/- s.e.m) ng progesterone/100 mg backfat were recorded on Day 15 of the oestrous cycle. It was estimated that, on this day, a total amount of about 36 mg progesterone is stored in the adipose tissue, which is approximately 200 times that present in total blood and corresponds to the daily production of the corpora lutea of the sow on Day 11. Initial half-life of progesterone in backfat after ovariectomy was estimated to be about 34 h compared to an initial half-life of plasma progesterone of about 120 min. The exact calculation of half-lives was, however, confounded by an obvious effect of anaesthesia or surgery on progesterone levels. Changes in backfat or plasma progesterone concentrations were not affected by the fat-to-lean ratio of the gilts. Fat progesterone levels determined in 44 additional pregnant and non-pregnant sows 17 or 20 days after mating indicated that reliable diagnosis of non-pregnant sows was possible on Day 20. It is concluded that the endocrinology of the oestrous cycle in pigs is related to the enormous storage of progesterone in the fat.     

Faecal steroid metabolites for non-invasive assessment of reproduction in common warthogs (Phacochoerus africanus), red river hogs (Potamochoerus porcus) and babirusa (Babyrousa babyrussa)

The objectives of this study were to analyse faecal steroid metabolites in African and South East Asian pig species kept in European zoos. Species studied were the warthog (Phacochoerus africanus), the red river hog (Potamochoerus porcus) and the babirusa (Babyrousa babyrussa). Faecal samples were collected 1-3 times per week from non-pregnant and pregnant captive female warthogs (n = 9), red river hogs (n = 7) and babirusas (n = 5). Enzyme-immunoassays for faecal progesterone, androgen, and oestrogen metabolites, were tested for their ability to determine follicular and luteal phases. In all three species, oestrous cycles could be monitored with 20alpha-OH- and 20-oxo-pregnane assays. In contrast, oestrogens and androgens were not useful in characterising follicular activity during the oestrous cycle in any species. Faecal 20alpha-OH- and 20-oxo-pregnane values were significantly correlated. Faecal pregnane concentrations revealed species-specific differences. Luteal phase values of 20alpha-OH-pregnanes were considerably higher than 20-oxo-pregnanes; 20alpha-OH-pregnanes were in the range of 3-10 microg/g in warthogs and red river hogs, whereas concentrations were 30-200 microg/g faeces in the babirusa. Regular oestrus cycles had a length of about 35 days in all three species studied. Results indicated a seasonal influence on the occurrence of reproductive cycles in the warthog with anoestrous periods in the European summer. The red river hog was found to be a seasonal and poly oestrous breeder; oestrus cycles started by January and continued until summer. In contrast, the babirusa showed non-seasonal ovarian cyclicity. In pregnant red river hogs, progesterone metabolites were comparable to luteal phase values of the oestrous cycle during the first 3 months of gestation, but did further increase during the last month of pregnancy. Oestrogens and 17-oxo-androstanes were significantly elevated during the second half of gestation. In summary, the reproductive biology of three exotic pig species was studied using non-invasive faecal steroid analysis and these methods were used for comparative investigations of oestrous cycles, pregnancy and seasonality.     

Oestradiol and progesterone receptors in the pig oviduct during the oestrous cycle

The concentrations of the tissue receptors for oestradiol (E) and progesterone (P) in the porcine oviduct at different stages on the oestrous cycle have been investigated by in vitro binding and exchange methods. Both hormones bound to specific cytoplasmic (Rc) and nuclear (Rn) receptor proteins with high affinity. The concentrations of ERc and ERn were two-fold higher in the ampulla as compared to the isthmus. The amount of ERc in the isthmic portion of the oviduct did not vary throughout the oestrous cycle. However, the ampullar ERc concentrations increased during prooestrus, showed a maximum at standing oestrus, thereafter decreasing. Significant variations in the amount of oviductal ERn were observed. Despite the differences in ERn amounts between segments, the concentration of ERn increased significantly during late prooestrus, attaining a three-fold elevation and remaining elevated during the period of standing oestrous and early luteal phase (days 3-4), thereafter returning to basal levels. No significant variations in the amount of isthmic PRc were found throughout the period studied. The ampulla, however, showed a significant increase in PRc concentrations during standing oestrus, thereafter decreasing. The concentrations of PRn in isthmus and ampulla were of about the same magnitude and varied significantly during the oestrous cycle, increasing in concentration from standing oestrous onwards. The temporal relationships between the variations in levels of oestradiol and progesterone receptors in oviductal tissues and those of the circulating plasma levels were established. The data obtained in this study suggest a relationship between the changes in the levels of oestradiol and progesterone oviductal binding during the first days of the oestrous cycle, and the gamete and embryo transport throughout the oviduct in the porcine species.     

Studies of the oestrous cycle, oestrus and pregnancy in the koala (Phascolarctos cinereus)

As an integral part of the development of an artificial insemination programme in the captive koala, female reproductive physiology and behaviour were studied. The oestrous cycle in non-mated and mated koalas was characterized by means of behavioural oestrus, morphology of external genitalia and changes in the peripheral plasma concentrations of oestradiol and progestogen. The mean (+/- SEM) duration of the non-mated oestrous cycle and duration of oestrus in 12 koalas was 32.9 +/- 1.1 (n = 22) and 10.3 +/- 0.9 (n = 24) days, respectively. Although the commencement of oestrous behaviour was associated with increasing or high concentrations of oestradiol, there were no consistent changes in the morphology or appearance of the clitoris, pericloacal region, pouch or mammary teats that could be used to characterize the non-mated cycle. As progestogen concentrations remained at basal values throughout the interoestrous period, non-mated cycles were considered non-luteal and presumed anovulatory. After mating of the 12 koalas, six females gave birth with a mean (+/- SEM) gestation of 34.8 +/- 0.3 days, whereas the remaining six non-parturient females returned to oestrus 49.5 +/- 1. 0 days later. After mating, oestrous behaviour ceased and the progestogen profile showed a significant increase in both pregnant and non-parturient females, indicating that a luteal phase had been induced by the physical act of mating. Progestogen concentrations throughout the luteal phase of the pregnant females were significantly higher than those of non-parturient females. Parturition was associated with a decreasing concentration of progestogen, which was increased above that of basal concentrations until 7 days post partum.     

The effect of trenbolone acetate on the bovine oestrous cycle

The anabolic steroid, trenbolone acetate, affected the bovine oestrous cycle in different ways depending on the timing of administration relative to oestrus and the plasma concentration of trenbolone achieved. Administration before oestrus caused failure of ovulation and a high incidence of a syndrome similar to cystic ovarian disease. High plasma concentrations of trenbolone during the luteal phase of the oestrous cycle prevented luteolysis and maintained progesterone secretion for up to 42 days. Subsequent cycles were not affected.     

Changes in phospholipids, cholesterol and protein content of oviduct fluid of cows during the oestrous cycle

The oviducts of 4 cows were cannulated and oviduct fluid was collected daily from the exteriorized cannulas for a total of 5 oestrous cycles. Daily serum samples were assayed for oestradiol-17 beta and progesterone to monitor the oestrous cycle. Data for each cycle were compared for oviduct fluid collected during the non-luteal phase (serum progesterone less than or equal to 1.5 ng/ml) and the luteal phase (serum progesterone greater than 1.5 ng/ml). During the non-luteal phase oviduct fluid volume was higher and the osmolality was lower than during the luteal phase. Total protein, cholesterol and phospholipid secreted daily was greater during the non-luteal phase. Cholesterol and protein concentrations were generally lower during the non-luteal phase, but phospholipid concentrations were generally higher. About 40% of the phospholipid in oviduct fluid was phosphatidylcholine and lysophosphatidylcholine, while phosphatidylinositol and lysophosphatidylinositol accounted for 20%. The ratio of 1-acyl-phospholipid to diacylphospholipid increased during the non-luteal phase. An increased cholesterol to phospholipid ratio, and a decreased cholesterol to protein ratio in oviduct fluid also were associated with the non-luteal phase. Changes in the lipid composition of oviduct fluid during the oestrous cycle may play a role in the preparation of gametes for fertilization.     

Plasma binding of oestradiol in progesterone-treated rats

Progesterone treatment of female rats causes an increase in body weight possibly via suppression of oestradiol secretion. This study was carried out to investigate the effect of progesterone on the non-protein bound and hence presumably biologically active fraction of oestradiol. Oestradiol binding to plasma proteins was studied in female Wistar rats during the oestrous cycle and after 12 days of treatment with progesterone (5 mg/day). There was no change in either the unbound fraction of oestradiol or plasma albumin concentrations during the oestrous cycle. Plasma oestradiol concentrations in progesterone-treated rats were similar to those seen during dioestrus, as were the degree of oestrogen binding and the plasma albumin concentrations. Although it was not feasible to calculate unbound concentrations, these results suggest that the increased body weight seen in progesterone-treated rats, and also during pregnancy, may be a result of suppression of unbound oestradiol concentrations to levels similar to those occurring during dioestrus.