cloprostenol and antarelix

The polypeptide ubiquitin covalently binds to cytoplasmic proteins and marks them for proteolytic degradation. Ubiquitin is upregulated during apoptosis in some systems. Apoptosis increases during luteolysis but it is not known whether ubiquitin is expressed in regressing corpora lutea. Marmoset ovaries were removed on day 10 of the luteal phase from animals that had received either no treatment, treatment with the PGF2 alpha analogue cloprostenol 24 h earlier, or treatment with the GnRH antagonist antarelix for either 24 or 48 h before ovary collection. Ubiquitin was localized on ovarian sections by immunocytochemistry, and oligonucleosome formation characteristic of apoptosis was examined in isolated corpora lutea by electrophoresis of extracted [32P]DNA. Oligonucleosome formation was low in midluteal corpora lutea on day 10 but increased after induced luteal regression with PGF2 alpha and GnRH antagonist. Nuclear ubiquitin immunoreactivity was found in 1.66 +/- 0.66 steroidogenic cells and cytoplasmic staining was found in 0.4 +/- 0.3 steroidogenic cells (per x 40 field of view) in midluteal phase corpora lutea on day 10. Luteolytic induction with PGF2 alpha significantly increased the number of cells exhibiting cytoplasmic immunoreactivity to 12.24 +/- 1.6 (P < 0.05). Ubiquitin immunoreactivity was not observed after GnRH-induced luteal regression. Apoptotic oligonucleosome formation was found after induced luteal regression with both PGF2 alpha and GnRH antagonist, but ubiquitin upregulation only occurred after PGF2 alpha-induced regression. These results indicate that ubiquitin expression is not specific for luteolysis and is not an indicator of luteal apoptosis, but that the polypeptide does play a role in luteal cellular responses to PGF2 alpha.

Topics: Animals; Apoptosis; Callithrix; Cloprostenol; Corpus Luteum; Dinoprost; Female; Gonadotropin-Releasing Hormone; Immunohistochemistry; Luteolysis; Oligopeptides; Ubiquitins

The mechanism controlling luteal regression in primates is unknown but may involve cell death by apoptosis. Marmoset ovaries containing corpora lutea were studied at different stages of the normal ovarian cycle. Two additional groups of animals underwent induced luteolysis with either the prostaglandin F2 alpha analogue, cloprostenol, or the GnRH antagonist, antarelix, at the mid-luteal phase. Apoptosis in ovarian sections was estimated both by counting the number of cells exhibiting morphological features of apoptosis and by in situ labelling the 3' ends of the DNA fragments with digoxigenin-11-dUTP. Apoptosis was found to be significantly increased in corpora lutea in the early follicular phase (equivalent to the later stage of luteal lifespan) compared with the mid-luteal phase corpora lutea, as judged by either computerized morphometry or 3' end labelling. Apoptosis was also increased by the administration of either cloprostenol or antarelix when using the 3' end labelling end point, but only after cloprostenol when using computerized morphometry. A further form of cell death, characterized by the formation of cytoplasmic vacuoles, was also observed in corpora lutea undergoing both induced and spontaneous regression. These results demonstrate that apoptosis within the primate corpus luteum is increased in both physiological and induced luteal regression. In addition, they show that an alternative form of cell death is involved in both spontaneous and induced luteal regression, although the relative importance of the two mechanisms remains to be determined.

Topics: Animals; Apoptosis; Callithrix; Cell Death; Cloprostenol; Corpus Luteum; Dinoprost; Female; Gonadotropin-Releasing Hormone; Luteolysis; Oligopeptides

There is increasing molecular evidence that apoptosis is involved in the process of structural luteal regression in non-primate species. Apoptosis is dependent upon the activation of certain proto-oncogenes and c-myc protein has an important regulatory role in this process in some cell types. The aim of the present study was to determine the occurrence and localisation of c-myc protein within the primate corpus luteum, determine changes during induction of luteal regression and examine the corpora lutea for morphological evidence of apoptosis. Ovaries were studied from marmoset monkeys in the late follicular, and in the early, mid and late luteal phases. Luteal regression was induced either by treatment with prostaglandin F2 alpha analogue or GnRH antagonist administered during the mid luteal phase and ovaries obtained 24 and 48 h later. Immunocytochemistry was performed using a monoclonal antibody to the c-myc protein. In pre-ovulatory follicles positive staining was found in the nucleus of a few granulosal cells and in the cytoplasm of thecal cells. c-myc was present in all corpora lutea where it was localised predominantly in the cytoplasm. In early corpora lutea, scattered cells with intense staining were observed in the presence of a majority of moderately or weakly stained cells. In the mid and late luteal phases, corpora lutea were uniformly moderately stained for c-myc. Following induction of luteal regression, regression, nuclear degeneration with condensation and fragmentation indicative of apoptosis was observed. In other luteal cells, increased membranes suggested necrosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Apoptosis; Callithrix; Cell Nucleus; Cloprostenol; Corpus Luteum; Cytoplasm; Female; Gonadotropin-Releasing Hormone; Hormone Antagonists; Immunohistochemistry; Luteal Phase; Luteolysis; Necrosis; Oligopeptides; Proto-Oncogene Proteins c-myc