CHANGES IN THE DENSITY AND PROGESTERONE CONTENT OF LUTEAL TISSUE IN THE EGYPTIAN BUFFALO DURING THE OESTROUS CYCLE

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.

1969 ◽  
Vol 43 (1) ◽  
pp. 1-NP ◽  
Author(s):  
A. S. EL-SHEIKH ◽  
F. B. SAKLA ◽  
S. O. AMIN

SUMMARY The histological and functional changes were studied in 30 corpora lutea (CL) of Egyptian buffaloes during four stages of pregnancy including the changes in shape, colour, weight and volume of CL, the volume of luteal cells and intercellular spaces, and in nuclear and cytoplasmic volume. Progesterone content and concentration were also estimated using column absorption chromatography and a counter-current distribution system. Active cells were numerous in the first two periods of gestation. Retrogression of the CL began in the third period of gestation as indicated by the decrease in active cells and the increase in degenerating luteal cells. The progesterone content and concentration in the CL increased consistently and reached its peak in the second period of pregnancy, it then decreased gradually until the fourth period.


1986 ◽  
Vol 113 (4) ◽  
pp. 570-575 ◽  
Author(s):  
Firyal S. Khan-Dawood

Abstract. Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and preoxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.


1997 ◽  
Vol 154 (2) ◽  
pp. 249-257 ◽  
Author(s):  
R K Arioua ◽  
C Féral ◽  
A Benhaïm ◽  
B Delarue ◽  
P Leymarie

Abstract It is well established that the rabbit corpus luteum (CL) function depends upon endogenous oestradiol, the major source of which in the rabbit ovary is considered to be the ovarian follicles. The absence of oestradiol formation by the rabbit CL has been previously reported. In a hyperstimulated pseudopregnant rabbit model used in our laboratory which developed a large number of corpora lutea in response to chorionic gonadotrophin (eCG)/hCG, we observed the survival of corpora lutea in vivo, and normal levels of plasma progesterone throughout pseudopregnancy (PP), despite the scarcity or the absence of follicles as a source of the luteotrophic hormone. Measurement of oestradiol in the plasma indicated that it was at high levels and correlated with the number of corpora lutea. This led us to investigate the luteal origin of oestradiol in this model. PP was induced in rabbits by i.m. injection of 200 IU eCG daily for 2 days followed on day 4 by i.m. injection of 200 IU hCG (day 0 of PP). Luteal tissue obtained at days 5, 9 and 12 of PP and cultured for 24 h synthesized oestradiol and testosterone in addition to progesterone. However, under the same conditions, follicles had limited capacity to secrete oestradiol. The presence of an aromatase activity in luteal tissue was confirmed when exogenous testosterone was added to the culture medium. P450aromatase (P450arom) mRNA was found in luteal tissue at days 5, 9 and 12 of PP. Small or large luteal cells, obtained by enzymatic digestion of the tissue followed by centrifugation in a Percoll density gradient, were cultured during several days with or without gonadotrophin or dibutyryl cAMP (dbcAMP). Both types of cells secreted oestradiol. In small cells and luteal tissue, aromatase activity was stimulated (1·5–2-fold) by hCG and dbcAMP. Large cells exhibited a greater capacity to aromatize testosterone than small cells, but aromatase activity was not modified by hCG or by dbcAMP. FSH had no effect on aromatase activity of either luteal cell type. This intrinsic luteal tissue aromatase capacity and the absence of premature regression of corpora lutea despite the limited support of follicular oestrogen, suggest an autocrine and luteotrophic role for this luteal oestrogen. Journal of Endocrinology (1997) 154, 249–257


2001 ◽  
Vol 73 (2) ◽  
pp. 323-327 ◽  
Author(s):  
Ş Arikan ◽  
A. Yigit

AbstractThis study was designed to investigate the size distribution of bovine steroidogenic luteal cells throughout pregnancy. Corpora lutea collected from three different stages of pregnancy were used. Luteal tissue was dissociated into single-cell suspension by enzyme treatments. Cells were stained for 3β-hydroxysteroid dehydrogenase (HSD) activity a marker for steroidogenic cells. The steroidogenic cells covered a wide spectrum of size ranging from 10 to 60 µm in diameter. There was a significant increase in mean cell diameter (P > 0·05) as pregnancy progressed. Mean diameter of 3β-HSD positive cells increased from 17·03 (s.e. 1·3) µm in the corpus luteum of early pregnancy to 33·38 (s.e. 2·4) µm in the corpus luteum of advanced pregnancy. The ratio of large (>22 µm in diameter) to small (10 to 22 µm in diameter) luteal cells was 0·32 : 1·0 in the early pregnancy, with the 10 to 22 µm cell size class predominant. However, the ratio of large to small luteal cells was increased to 6·49 : 1·0 µm as pregnancy advanced and 23 to 42 µm cell sizes become predominant. It is likely that small luteal cells develop into large cells as gestation progresses. Development of pregnancy is associated with an increase in size of steroidogenic luteal cells.


Reproduction ◽  
2000 ◽  
pp. 109-114 ◽  
Author(s):  
Z Ge ◽  
WE Nicholson ◽  
DM Plotner ◽  
CE Farin ◽  
JE Gadsby

Insulin-like growth factor I (IGF-I) is believed to play a luteotrophic role in the pig corpus luteum during the oestrous cycle. Since the actions of IGF-I in target tissues are mediated by the type I IGF receptor, the concentrations of IGF-I receptor mRNA and protein were examined in pig corpora lutea at different stages of the oestrous cycle. Corpora lutea were collected from normally cyclic gilts on days 4, 7, 10, 13, 15 and 16 of the oestrous cycle (n = 4 animals per day). Corpora lutea on days 7, 10 and 13 were dissociated with collagenase, and large and small luteal cell sub-populations were separated by elutriation. Northern and slot blots were used to examine mRNA, and western blots were used to measure the concentrations of IGF-I receptor protein in the pig corpus luteum. On northern blots, luteal IGF-I receptor mRNA was present as a single 11 kb transcript. The slot blots showed that the steady state expression of IGF-I receptor mRNA increased significantly (P < 0.05) from its lowest value on day 4, to reach a maximum on days 13-16. IGF-I receptor mRNA was also expressed to a greater extent in large compared with small luteal cells (P < 0.05). On western blots, IGF-I receptor appeared as a 95 kDa protein band (beta-subunit) and IGF-I receptor protein concentrations were significantly higher (P < 0.05) on days 4-10 than on days 13-16. Finally, large luteal cells appeared to contain more IGF-I receptor protein than the small luteal cells. In conclusion, since IGF-I receptor was detected in the pig corpus luteum, it is a likely target tissue for IGF-I, especially during the early luteal phase. Furthermore, IGF-I receptor was localized primarily on large luteal cells, thus it is hypothesized that IGF-I may play a paracrine role in the pig corpus luteum.


Endocrinology ◽  
1997 ◽  
Vol 138 (8) ◽  
pp. 3236-3241 ◽  
Author(s):  
G. Thordarson ◽  
S. Galosy ◽  
G. O. Gudmundsson ◽  
B. Newcomer ◽  
R. Sridaran ◽  
...  

Abstract Pituitary hormones are essential for the maintenance of the corpus luteum in the pregnant mouse during the first half of gestation. Thereafter, hormones from the placenta take over the luteotropic role of the pituitary hormones. Mouse placental lactogen-I (mPL-I) and mPL-II, two PRL-like hormones produced in the placenta, are probably necessary for the maintenance of the corpus luteum in the latter half of pregnancy. A culture system of luteal cells from pregnant mice was developed to investigate the role of hormones from the placenta that may be important for the function of the corpus luteum. Mice were killed on days 10, 14, and 18 of pregnancy, and the corpora lutea were excised from the ovaries and digested in 0.1% collagenase, 0.002% DNase for 1 h. The resulting luteal cell suspension was plated onto 96-well plates coated with fibronectin (1 × 105 cells/well) and cultured for 1–3 days. Medium was changed daily. The cells were treated with various concentrations and combinations of mPL-I, mPL-II, mouse PRL, androstenedione, dihydrotestosterone, 17β-estradiol (E2), testosterone, hydroxyflutamide, cycloheximide, actinomycin D, and fadrozole to study the effects of these different treatments on progesterone (P4) production. The three lactogens (mPL-I, mPL-II, and mouse PRL) all stimulated the release of P4 from the luteal cells. The potency of the lactogens was similar and did not depend on the stage of pregnancy at which the luteal tissue was obtained. However, the responsiveness of the cells to all hormone-stimulated P4 release was gradually reduced the later in pregnancy the tissue was collected. Androgens also stimulated the release of P4 from the luteal cells, and when administered together, the lactogens and the androgens acted synergistically to stimulate P4 release. The androgens acted directly but not through conversion to E2, as determined by the findings that 1) the effects of the androgens could not be reproduced by E2 administration, 2) nonaromatizable androgen dihydrotestosterone was as effective as aromatizable androgens, and 3) aromatase inhibitor did not prevent the action of the androgens to stimulate the P4 release. The effect of the androgens on the P4 release was rapid, occurring within 15 min of hormone administration. It was not prevented by inhibitors of protein and RNA synthesis, and the intracellular androgen receptor antagonist hydroxyflutamide did not affect the androgen action. Therefore, the androgen effects were not mediated through the intracellular androgen receptor and de novo protein synthesis was not needed for androgen-stimulated P4 release.


1989 ◽  
Vol 121 (1) ◽  
pp. 117-123 ◽  
Author(s):  
C. Sernia ◽  
R. T. Gemmell ◽  
W. G. Thomas

ABSTRACT There is inconclusive evidence that oxytocin acts directly on the corpus luteum and affects steroidogenesis. Since any such action would probably be mediated by oxytocin receptors, these should be present in luteal tissue. In this study, homogenates of corpora lutea from both pregnant and non-pregnant ewes were examined for oxytocin receptors by radio-receptor assay. Specific oxytocin binding was not observed in luteal tissue during the oestrous cycle. However specific binding was found in the corpora lutea of pregnant ewes; appearing at a fetal head length of approximately 0·65 cm (about 30 days of pregnancy) and persisting to a head size of 11 cm, the largest size examined in this study. The affinity (Kd) of the receptor was calculated as 2·9 ± 0·3 nmol/l (s.e.m.; n = 9), a value similar to that obtained for the uterus. The receptor number ranged from a low of 8·7± 3·2 fmol/mg protein (n = 6) at a head size of <0·65 cm, to a maximum of 40·1 ± 6·5 fmol/mg protein (n = 25) at a head size of 2·5–3·75 cm. These values were lower than our estimate of 588 ± 39 fmol/mg protein (n = 5) for the uterus. It is concluded that a direct action of oxytocin on the corpus luteum is possible but only after the first month of pregnancy and not in the corpus luteum of the oestrous cycle. Journal of Endocrinology (1989) 121, 117–123


2008 ◽  
Vol 20 (1) ◽  
pp. 237
Author(s):  
L. G. B. Siqueira ◽  
J. H. M. Viana ◽  
C. A. A. Torres ◽  
E. D. Souza ◽  
L. S. Amorim ◽  
...  

It has been suggested that ultrasound image attributes are a potential indicator of the physiological and functional status of the corpus luteum (CL). The aim of this study was to evaluate corpus luteum morphological and echotextural changes, and to correlate these changes with plasma progesterone concentration [P4] throughout the bovine estrous cycle. Crossbred heifers were scanned daily, using a B-mode, real-time ultrasound machine equipped with a 5-MHz linear-array rectal transducer, throughout a natural estrous cycle (Experiment 1; n = 12) or during a shorter estrous cycle, interrupted on the 10th day, by luteolysis induction (Experiment 2; n = 6). Blood samples were collected for further plasma [P4] analyses by RIA. Corpora lutea areas (cm2) were measured, and daily images of each CL were videotaped (VHS tapes) until digitized. Computer-assisted analyses of image attributes were performed using a custom-developed software. Daily values of luteal area, echotexture, and plasma [P4] values were analyzed by ANOVA with Tukey's test to determine differences among means of each cycle day. Pearson's correlation coefficients were calculated between luteal area, mean pixel value, pixel heterogeneity, and plasma [P4]. In the first experiment, luteal tissue area increased to a maximum on the 10th day (P < 0.05), followed by a plateau, and then declined from Day 14 to next estrus. There was a significant correlation between luteal tissue area and plasma P4 (r = 0.69; P < 0.01). In the second experiment, plasma P4 dropped to basal values 24 h after luteolysis induction. Luteal tissue area decreased at a slow rate, and reached values similar to ones from metestrus 36 h after treatment. In Experiment 1, echotexture parameters of the CL were analyzed after data adjustment to the onset of luteolysis. In both experiments, mean pixel values did not change throughout the estrous cycle and there was no correlation between mean pixel values and plasma [P4] (P > 0.10). Pixel heterogeneity changed throughout the natural estrous cycle, with maximum value on metestrus (Day 14; Day 0 = luteolysis) and minimum on diestrus (Day 2; P < 0.01). However, this parameter did not change when luteolysis was induced (Experiment 2; P > 0.10). There were significant correlations between pixel heterogeneity and plasma progesterone in both of the experiments (r = –0.69 and r = –0.48; P < 0.05). In conclusion, mean pixel values do not reflect morphological or functional changes of the CL throughout the estrous cycle. On the other hand, based on the correlations between pixel heterogeneity and systemic [P4] in both experiments, this image attribute (heterogeneity) has the potential to indicate functionality and steroidogenic capacity of the luteal gland.


Author(s):  
Emilia Przygrodzka ◽  
Corrine F. Monaco ◽  
Michele R. Plewes ◽  
Guojuan Li ◽  
Jennifer R. Wood ◽  
...  

In the absence of pregnancy the ovarian corpus luteum undergoes regression, a process characterized by decreased production of progesterone and structural luteolysis involving apoptosis. Autophagy has been observed in the corpus luteum during luteal regression. Autophagy is a self-degradative process important for balancing sources of cellular energy at critical times in development and in response to nutrient stress, but it can also lead to apoptosis. Mechanistic target of rapamycin (MTOR) and 5′ AMP-activated protein kinase (AMPK), key players in autophagy, are known to inhibit or activate autophagy, respectively. Here, we analyzed the signaling pathways regulating the initiation of autophagy in bovine luteal cells. In vivo studies showed increased activating phosphorylation of AMPKα (Thr172) and elevated content of LC3B, a known marker of autophagy, in luteal tissue during PGF2α-induced luteolysis. In vitro, AMPK activators 1) stimulated phosphorylation of regulatory associated protein of MTOR (RPTOR) leading to decreased activity of MTOR, 2) increased phosphorylation of Unc-51-Like Kinase 1 (ULK1) and Beclin 1 (BECN1), at sites specific for AMPK and required for autophagy initiation, 3) increased levels of LC3B, and 4) enhanced colocalization of autophagosomes with lysosomes indicating elevated autophagy. In contrast, LH/PKA signaling in luteal cells 1) reduced activation of AMPKα and phosphorylation of RPTOR, 2) elevated MTOR activity, 3) stimulated phosphorylation of ULK1 at site required for ULK1 inactivation, and 4) inhibited autophagosome formation as reflected by reduced content of LC3B-II. Pretreatment with AICAR, a pharmacological activator of AMPK, inhibited LH-mediated effects on RPTOR, ULK1 and BECN1. Our results indicate that luteotrophic signaling via LH/PKA/MTOR inhibits, while luteolytic signaling via PGF2α/Ca2+/AMPK activates key signaling pathways involved in luteal cell autophagy.


1968 ◽  
Vol 59 (2_Suppl) ◽  
pp. S35-S51 ◽  
Author(s):  
B. L. Lobel ◽  
E. Levy

ABSTRACT Activities of various hydrolases and dehydrogenases were studied during the formation, development and involution of cyclic corpora lutea and in the corpora lutea of early pregnancy. At 24 hours postovulation the luteal cells, whether of granulosal or thecal origin, contained demonstrable levels of Δ5-3β-hydroxysteroid dehydrogenase and the NADP and NADPH2 diaphorases. During the period of proliferation and cellular growth, enzymic activities in the luteal cells were moderate at first, and then increased. In the mature corpus luteum, activities of the dehydrogenases occurred in all luteal cells but were most intense in the large polymorphic luteal cells. Activities of hydrolytic enzymes, low in the immediate postovulatory period, increased with the development of the vascular system. Enzymic characteristics of corpora lutea of gestation were similar to those of cyclic corpora, except for phosphorylase activity which was observed in luteal cells in gestational corpora, but confined to the vascular walls in cyclic corpora. No increase in activities of 17β- and 20β-hydroxysteroid dehydrogenases (above those seen in pre-ovulatory follicles) were observed after incubation of sections of either mature cyclic or gestational corpora. Involution of cyclic corpora lutea began with degenerative changes in the blood vessels: pyknosis of the endothelial cell nuclei and a sudden decline in activities of hydrolytic enzymes in the vascular walls. Subsequently, the luteal cells showed a sharp decrease in activities of the dehydrogenases as well as other signs of regressive change. The cytochemical findings are discussed in relation to biochemical observations on steroid synthesis by the bovine corpus luteum.


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