IDENTIFICATION AND QUANTITATIVE DETERMINATION OF STEROIDS IN BOVINE CORPUS LUTEUM DURING OESTROUS CYCLE AND PREGNANCY

1975 ◽  
Vol 80 (1) ◽  
pp. 149-164 ◽  
Author(s):  
M. Axelson ◽  
G. Schumacher ◽  
J. Sjövall ◽  
B. Gustafsson ◽  
J. O. Lindell

ABSTRACT Neutral steroids in bovine corpus luteum were isolated by liquid-gel chromatography on hydrophobic Sephadex, and were analyzed by computerized gas chromatography-mass spectrometry. The presence of progesterone and 20β-hydroxy-4-pregnen-3-one was confirmed. In addition, 3β- hydroxy- 5- pregnen -20- one, 5- pregnene- 3β,20β -diol, 3β-hydroxy-5α-pregnan-20-one and 5α-pregnane-3β,20β-diol,32 were fully identified, and 3-hydroxy-4-pregnen-20-onne, 4 pregnene-3,20-diol, 22-hydroxycholesterol and 20,22-dihydroxycholesterol were partially characterized. Steroid sulphates were not detected. Quantification of the six fully identified steroids was based on peak areas in specific fragment ion current chromatograms constructed by the computer. During the 4th–19th day of the oestrous cycle the steroid concentrations varied as follows: progesterone 6.0–36.7 μg/g wet luteal weight, 20β-hydroxy-4-pregnen-3-one 0.8–5.5 μg/g, 3β-hydroxy-5-pregnen-20-one 1.0–7.1 μg/g, 5-pregnene-3β,20β-diol < 0.2–0.9 μg/g, 3β-hydroxy-5α-pregnan-20-one 1.7–8.6 μg/g, and 5α-pregnane-3β,20β-diol < 0.2-1.2 μg/g. The concentrations of progesterone and 3β-hydroxy-5-pregnen-20-one seemed to vary in parallel and were low during days 11–17. During this period the concentrations of 5-pregnene-3β,20β-diol and 5α-pregnane-3β,20β-diol were highest as was the relative contribution of all three 20β-hydroxysteroids to the total amount of steroids. The relative amount of 3β-hydroxy-5α-pregnan-20-one seemed to be highest during days 4–6. The total steroid concentration in corpora lutea taken in early pregnancy (75–105 days) was 18–47 μg/g. In the period 75–90 days, progesterone constituted only 35–42 % of the total steroids, 3β-hydroxy-5α-pregnan-20-one as much as 23–40 % and the total 20β-hydroxysteroids 18–30%. The total steroid concentration in corpora lutea taken in midterm and late pregnancy was 21–77 μg/g. In this period progesterone was by far the predominant steroid and constituted about 80–90 % of the total steroids in corpora lutea taken between days 150 and 240. Possible correlations between luteal growth, steroid oxidoreductases and steroid concentrations are discussed.

1972 ◽  
Vol 69 (2) ◽  
pp. 369-384
Author(s):  
H. Brandau ◽  
L. Brandau ◽  
G. Mutzke

ABSTRACT In the bovine corpora lutea periodical activities of the Δ53β-, 3β-, 17β-and 20β-hydroxysteroid dehydrogenase (OHSDH) as well as activities of glyceraldehyde-3-phosphate- and glucose-6-phosphate dehydrogenase were measured quantitatively and the alterations throughout the different stages of the cycle were studied. After homogenization of the tissue and fractionate centrifugation the enzyme activities were determined by a standardized optical method. The activities of the Δ53β-, and 3β- and 17β-OHSDH increase slowly during the first 7 days of the cycle, the maximum is reached abruptly on the 12th to 13th day of the cycle. After a striking reduction the activities decline continually to the 19th to 21st day reaching the values detected at the beginning of the cycle. The 20β-OHSDH increases slowly to the maximum on the 15th day of the cycle. Activities of the 3α-OHSDH were obtained only inconsistently. The behaviour of the activities of G6PDH was nearly identical with that of the 3β-OHSDH, while the GAPDH shows only little fluctuations of its activities. The obtained enzyme activity patterns of the maturating and high functional corpus luteum correspond to the well-known data of the biosynthetic function of the bovine corpus luteum. The changes of the amounts of progesterone and 20β-progesterol agree with the course of the activities of the 3β- resp. 20β-OHSDH.


1995 ◽  
Vol 7 (3) ◽  
pp. 303 ◽  
Author(s):  
RT Gemmell

The corpus luteum (CL) is a transitory organ which has a regulatory role in reproduction. Sharks, amphibians and reptiles have corpora lutea that produce progesterone which influences the rate of embryonic development. The egg-laying monotremes and the two major mammalian groups, eutherian and marsupial, have a CL that secretes progesterone. Most eutherians have allowed for the uterine development of their young by extending the length of the oestrous cycle and the CL or placenta actively secretes progesterone until birth. Gestation in the marsupial does not extend beyond the length of an oestrous cycle and the major part of fetal development takes place in the pouch. Where the extension of the post-luteal phase in the eutherian has allowed for the uterine development of young, the marsupial has extended the pre-luteal phase of the oestrous cycle and has evolved an alternative reproductive strategy, embryonic diapause. The mechanism for the secretion of hormones from the CL has been controversial for many years. Densely-staining secretory granules have been observed in the CL of sharks, marsupials and eutherians. These granules have been reported to contain relaxin, oxytocin or mesotocin, and progesterone. A hypothesis to suit all available data is that all hormones secreted by the CL are transported within such granules. In conclusion, although there are obvious differences in the mode of reproduction in the two main mammalian groups, it is apparent that there is a great deal of similarity in the hormonal control of regression of the CL and parturition.


1996 ◽  
Vol 1996 ◽  
pp. 69-69
Author(s):  
AJ Holt ◽  
RG Rodway ◽  
JBC Findlay ◽  
HS Sands ◽  
DN Batchelder

The role of β-carotene in the fertility of ruminant animals has long been acknowledged (Friesecke, 1978). Initially this was thought to be due to its action as a vitamin A precursor but recently β-carotene itself has been considered responsible for improving reproductive performance (Hurley & Doane, 1989). The mechanism by which β-carotcne acts is unclear, but as its concentration in the corpus luteum was typically found to be 70μg per gram of tissue, its biological action is probably exerted in this area.β-Carotene has been studied in the bovine corpus luteum using Raman spectroscopy, high performance liquid chromatography (HPLC) and gel filtration chromatography. The structure of β-carotene produces a characteristic Raman spectrum and by utilising an imaging technique, photographs of isolated luteal cells were obtained indicating the regions of β-carotene within them. Differential centrifugation was used to obtain pure subcellular fractions of luteal cells.


1967 ◽  
Vol 39 (2) ◽  
pp. 163-171 ◽  
Author(s):  
A. S. EL-SHEIKH ◽  
FRANÇOIS B. SAKLA ◽  
SAFAA O. AMIN

SUMMARY The histological and functional changes of 31 corpora lutea of Egyptian buffaloes during the various phases of the oestrous cycle were studied. The volumes of the corpora lutea were calculated, the volume per cell, the cell volume and the volume of the intercellular spaces were estimated from transverse serial sections stained with haematoxylin and eosin, Mallory's triple stain or van Gieson's stain. The nuclear volumes were also determined and the cytoplasmic volume was calculated. The progesterone content was estimated using column absorption chromatography and a counter-current distribution system. It was concluded that the luteal cells increase both in volume and in number due to mitosis. The luteal cells decrease in volume after the 15th day after ovulation, the cells lose their distinct outlines in the regressive stage and disappear completely in the corpus albicans. There was a parallel increase in luteal cell volume and progesterone content until the 15th post-ovulatory day followed by a decrease in the regressive phase and disappearance of the hormone in the corpus albicans. A highly significant correlation (r = +0·875) was found between the progesterone content and the cytoplasmic volume. Progesterone concentration/g. luteal tissue increased from the corpus haemorrhagicum to the mature corpus luteum, decreased in the regressive corpus luteum and completely disappeared in the corpus albicans.


1979 ◽  
Vol 82 (3) ◽  
pp. 425-428 ◽  
Author(s):  
J. WATSON ◽  
C. E. PATEK

Prostaglandin F2α (PGF2α) secreted by the reproductive tissues of the pig in vitro was measured and it was found that the levels secreted by the corpus luteum and endometrium of early pregnant sows were significantly lower than those secreted by tissues during the late stage of the oestrous cycle. They were, however, comparable to levels secreted by tissues from the mid-stage of the oestrous cycle. Embryos also secreted significant amounts of PGF2α. Secretion of progesterone and oestradiol by the corpora lutea of both cyclic and pregnant pigs fell within accepted limits but embryos were also found to secrete significant amounts of oestradiol. The results suggest that luteal maintenance in the early pregnant pig is unlikely to be directly due to reduced synthesis of PGF2α.


1976 ◽  
Vol 70 (1) ◽  
pp. 39-45 ◽  
Author(s):  
F. R. BLATCHLEY ◽  
B. T. DONOVAN

SUMMARY The response of the guinea-pig corpus luteum to the luteolytic influence of glass beads placed in the uterus, or to prostaglandin administration, was followed by assay of the progesterone content of blood samples collected daily. Following the introduction of glass beads into the uterus early in the cycle, the secretion of progesterone was curtailed. Treatment with prostaglandin F2α over days 4–6 or 6–8 of the cycle temporarily depressed progesterone release without shortening the life of the corpora lutea. When the drug was administered over days 8–10, 10–12 or 12–14 the depression in progesterone was not followed by any recovery. These observations indicate that the response of the corpora lutea to a luteolytic influence changes during the oestrous cycle.


1971 ◽  
Vol 50 (4) ◽  
pp. 625-635 ◽  
Author(s):  
DOREEN V. ILLINGWORTH ◽  
J. S. PERRY

SUMMARY The effects of hypophysial stalk-section on the growth and function of the corpus luteum of the non-pregnant guinea-pig have been compared with the effects of hypophysectomy (as previously described) and with the effects of prolactin administered to hypophysectomized animals. Stalk-section soon after ovulation did not impair the growth of the corpora lutea nor their ability to secrete progesterone. Stalk-section before day 9 of the oestrous cycle prevented the normal regression of the corpora lutea; they continued to grow and 3 weeks after ovulation were as large as those of pregnant animals, or of non-pregnant hysterectomized guinea-pigs. The corpora lutea regressed irregularly during the following 2 weeks. When performed on, or later than day 9, stalk-section did not prevent luteal regression at the normal time. Administration of prolactin (10 i.u./day) to hypophysectomized guinea-pigs restored the growth-rate of the corpora lutea, which reached sizes comparable to those of the normal cycle, and those of stalk-sectioned animals, by 10 days after ovulation. Our results indicate that prolactin can have substantial luteotrophic activity in the guinea-pig.


2002 ◽  
Vol 75 (3) ◽  
pp. 427-432 ◽  
Author(s):  
Ş. Arikan ◽  
A. A. Yigit

AbstractThe present study examines the size distribution of ovine steroidogenic and non-steroidogenic luteal cells throughout pregnancy. Cells were isolated from corpora lutea collected from early (< 8 weeks), mid (9 to 14 weeks) or late (15 to 18 weeks) stages of pregnancy. Cells were stained for 3β-hydroxysteroid dehydrogenase (3β-HSD) activity, a marker for steroidogenic cells. Both 3β-HSD positive and β-HSD negative cells covered a wide spectrum of size ranging from 7 to 37 μm in diameter. There was a significant increase (P > 0·01) in mean diameter of non-steroidogenic luteal cells as pregnancy progressed. Mean diameter of 3β-HSD negative cells increased from 17·8 (s.e. 0·4) μm in the corpus luteum of early stage of pregnancy to 22·4 (s.e. 0·3) μm in the corpus luteum of advanced pregnancy. However, there was no significant increase in the mean diameter of 3β-HSD positive cells. Corpora lutea obtained from early stages of the pregnancy contained more steroidogenic cells than the cells obtained from mid and late pregnancy (P < 0·01). Percentage of 3β-HSD negative cells had increased 2·07-fold by 18 weeks of pregnancy when compared with the early stage of pregnancy. In contrast, percentage of 3β-HSD positive cells had decreased to 50% of starting values during the same period (P < 0·05). These results indicate that the ovine corpus luteum of pregnancy is morphologically dynamic over the course of pregnancy. Steroidogenic activity of luteal cells may decrease as pregnancy progresses, especially activity of the large luteal cells.


1973 ◽  
Vol 58 (3) ◽  
pp. 493-501 ◽  
Author(s):  
E. R. SIMPSON ◽  
C. R. JEFCOATE ◽  
R. A. COLLETT ◽  
R. B. LAND

SUMMARY Cytochrome P-450 of mitochondria from the ovine corpus luteum was examined on different days throughout the oestrous cycle. Most, if not all, of the mitochondrial cytochrome P-450 appeared to be of the type involved in cholesterol side-chain cleavage, as determined by steroid binding spectra. The content of mitochondrial cytochrome P-450 in the corpus luteum changed considerably throughout the cycle, and closely paralleled the rise and fall of plasma progesterone concentration; the correlation between these two parameters was 0·96. This was largely a reflection of the changes in weight of the corpus luteum throughout the cycle, since the specific content of cytochrome P-450 changed much less dramatically. The percentage of the mitochondrial cytochrome P-450 present as the cholesterol—substrate complex remained constant throughout the cycle at about 25–30% of the total.


1989 ◽  
Vol 120 (2) ◽  
pp. 325-330 ◽  
Author(s):  
J. Th. J. Uilenbroek ◽  
P. J. A. Woutersen ◽  
P. D. M. van der Vaart

ABSTRACT Corpora lutea could be identified under the dissection microscope up to 7 days after formation. They were isolated during the oestrous cycle and pseudopregnancy and the progesterone and 20α-OH-progesterone contents were compared with serum values of these steroids. The pattern of progesterone in serum resembled that found in the corpora lutea. However, the pattern of 20α-OH-progesterone concentrations in serum and corpora lutea were different. While 20α-OH-progesterone concentrations in the corpora lutea showed large variations during the cycle, changes in serum concentrations of 20α-OH-progesterone were relatively small. Measurement of hormone concentrations in isolated corpora lutea is therefore a sensitive method for studying corpus luteum activity. To study whether corpora lutea derived after ovulation of immature follicles showed deficient luteal activity, rats at dioestrus (2 days before pro-oestrus) were induced to ovulate by the injection of 10 IU human chorionic gonadotrophin (hCG) and subsequent luteal activity was studied by measuring hormone concentrations in the corpora lutea on day 5 of pseudopregnancy. Concentrations of progesterone, but not of 20α-OH-progesterone, in corpora lutea derived from follicles induced to ovulate at dioestrusday 1 were significantly lower than those in corpora lutea derived from follicles induced to ovulate at prooestrus. This difference was observed not only when pseudopregnancy was induced by cervical stimulation but also when it was induced by implantation of a pituitary gland under the kidney capsule. However, in the latter case, corpora lutea already present on the day of hCG injection also became activated. The present experiments demonstrate that by measuring hormone concentrations in isolated corpora lutea changes in luteal activity can be studied effectively. Moreover, it appears that corpora lutea derived from immature follicles contained less progesterone than those derived from fully mature follicles. Journal of Endocrinology (1989) 120, 325–330


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