Inducing cell death in human carcinoma cells with beta-lapachone revealed the presence of caspase-3 in apoptotic cell death, but not in necrotic cell death (Li et al.,1999). Caspase-3 is activated by caspase-8. Activation of caspase is preceded by the release of mitochondrial cytochrome (Li et al.,1999).
NCBI Summary:
This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 6, 7 and 9, and the protein itself is processed by caspases 8, 9 and 10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. Alternative splicing of this gene results in two transcript variants that encode the same protein. [provided by RefSeq, Jul 2008]
Caspase-3 is the principal downstream effector of cell death (Boone et al.,1998). After inducing apoptosis in mouse ovary granulosa cells, Robles et al.(1999) found that healthy granulosa cells possess almost exclusively the inactive pro-caspase-3 form whereas granulosa cells undergoing apoptosis activated by Apaf-1 (by depriving cells of gonadotropins to suppress Apaf-1) rapidly process procaspase-3 to the active enzyme.
Cellular localization
Cytoplasmic, Nuclear
Comment
Ovarian function
Follicle atresia, Luteolysis
Comment
Caspase-3 gene expression in human luteinized granulosa cells is inversely correlated with the number of oocytes retrieved after controlled ovarian stimulation. Lobach VN et al. (2017) Granulosa cells control oocyte maturation through paracrine signalling and changes to the microenvironment around the oocyte. Apoptosis occurs as a physiological mechanism of granulosa cell renewal, but how it relates with the ovarian response to induced ovulation is still unclear. Therefore, this study evaluated apoptosis-related gene expression levels in granulosa cells of patients undergoing controlled ovarian stimulation. We enrolled prospectively 59 consecutive IVF patients referred to a tertiary academic hospital for couple infertility treatment. Luteinized granulosa cells were isolated from follicular fluid and the RNA was extracted, reverse-transcribed and the gene expression of apoptosis inducers (caspase-3, caspase-8 and bax) and inhibitor (Bcl-2) was quantified by real-time polymerase chain reaction. Caspase-3 gene expression correlated negatively with the number of pre-ovulatory follicles (Spearman's r = -0.308), the number of collected oocytes (r = -0.451), the number of mature oocytes (r = -0.526), the number of fertilized oocytes (r = -0.439) and the number of viable embryos (r = -0.443, all statistically significant at p < 0.02 level). No such associations were found with caspase-8, bax or bcl-2. These preliminary findings suggest that increased caspase-3 gene expression in granulosa cells is associated with a worse ovulatory response in humans.//////////////////
Expression of Caspase-3 detected in follicular cells of atretic follicles, and in interstitial cells, in surface epithelial and vascular endothelial cells, and in some thecal cells of post-ovulatory follicles of the quail ovary (Van Nassauw et al.,1999).
Apoptosis during PGF2alpha-induced luteal regression in ewes showed 3-fold higher levels of caspase-3 in the corpus luteum than untreated ewes (Rueda et al., 1999).
Activity and expression of different members of the caspase family in the rat corpus luteum (CL) during pregnancy and postpartum. Peluffo MC et al. Studies were designed to examine the expression and activity of four caspases, which contribute to the initial (caspases-2, -8 and -9) and final (caspase-3) events in apoptosis, in the rat CL during pregnancy (days, 7, 17, 19 and 21 of gestation), postpartum (day 1 and 4) and after injection (0, 8, 16, 24 and 36 h) of the physiological luteolysin, PGF-2alpha. In addition, the temporal relationship of caspase expression/activity relative to steroid production and luteal regression was evaluated. During pregnancy, the activity of all four caspases was significantly greater on day 19, prior to a decline in CL progesterone (P) and CYP11A1 levels at day 21 of gestation. The levels of the caspase-3 active fragment (p17, measured by Western blot) also increased at day 19 and 21 of pregnancy. Immunohistochemical analyses detected specific staining for the caspases in luteal cells (large and small), as well as, in endothelial cells. However, the percentage of apoptotic cells did not increase in the CL until postpartum. Following PGF-2alpha injection, there was a significant decrease in CL P by 24 h, though the activity of all four caspases did not increase until 36 h post treatment. The active p17 fragment of caspase-3 also significantly increased at 36 h post- PGF-2alpha. These results suggest that an increase in the activity of caspase-2, -8, -9 and -3 is associated with the early events of natural luteolysis at the end of pregnancy. Also the exogenous administration of the luteolysin PGF-2alpha may regulate members of the caspase family. Key words: caspases, luteolysis, corpus luteum, pregnancy.
Expression regulated by
FSH, LH
Comment
Presence of caspase-3 in granulosa cells of atretic, but not healthy, follicles suggests that the expression of this enzyme is regulated by gonadotropin and may be up-regulated as part of the apoptotic process in granulosa cells (Boone et al.,1998).
Apoptosis causes the elimination of ovarian germ cells and the atretic degeneration of ovarian follicles. Yacobi K, et al 2004 have used cultured rat preovulatory follicles to examine the regulation of effector caspase-3 and -7 in follicles undergoing apoptosis in the presence or absence of gonadotropins or IGF-I. Culturing follicles in the presence or absence of serum resulted in the induction of apoptosis of granulosa cells (GC), which was accompanied by effector caspase activation. Surprisingly, the addition of the survival factors LH or FSH, but not IGF-I, further increased caspase-3 and -7 activity. Immunohistochemistry studies of the LH and FSH treated follicles indicated that cleaved caspase-3 was predominantly localized to the peripheral theca-interstitial cells (TIC). Western blot analysis and caspase-3 and -7 activity assays of the separated follicular compartments confirmed that both LH and FSH treatments significantly enhance caspase-3 and -7 activity in TIC. The elevation in caspase-3 and -7 activity in TIC was accompanied by an increase in apoptosis. Interestingly, LH and FSH also induced an increase in caspase-3 and -7 activity in GC, however this increase was accompanied by a decrease in apoptosis. Finally, we demonstrate that in freshly isolated preovulatory follicles and in antral follicles in intact ovaries, the expression level of pro-caspase-3 is significantly higher in TIC than in GC. Thus, LH and FSH have a dual effect on the cultured rat preovulatory follicle: an anti-apoptotic effect on GC and a pro-apoptotic effect on TIC.
Expression of caspase-1, caspase-3, DNA fragmentation factor, and apoptotic protease activating factor-1, which are intermediate molecules in apoptotic pathways, was demonstrated in
granulosa cells from patients undergoing IVF. Also, proforms, but not activated enzymes, for both caspase-1 and caspase-3 were observed (Izawa et al.,1998). Caspase-3 found to be localized in luteal cells of healthy corpora lutea and in theca, but not in granulosa cells of healthy follicles; however, caspase-3 was found to be expressed in granulosa cells of atretic follicles in a pattern similar to that of the localization of granulosa cell death (Boone et al.,1998). Consistent with the caspase-3 role in apoptosis, staining in human tissue found high intensity of caspase-3 in corpus luteal cells, but low intensity in follicular granulosa cells in the ovary (Krajewska et al.,1997).
Follicle stages
Secondary, Antral, Preovulatory, Corpus luteum
Comment
Fenwick MA, et al 2002 .
investigated the expression of caspase-3 in the mouse ovary, and determined whether active caspase-3 is present within smaller follicles, which may constitute the resting pool. The inactive enzyme was expressed as a 32 kDa band on a western blot of tissue extracts, whereas the active form was localized immunohistochemically. Bromodeoxyuridine (BrdU) was administered to mice (n = 7) during a 12 h period and subsequently localized to identify potentially quiescent follicles. Measurements of BrdU-positive cells in the mouse ovary were extrapolated with data obtained by morphometric analyses of small follicles using the nucleator technique. BrdU was incorporated into the granulosa cells of follicles regardless of size and the number of cells they contained, but was absent in a large proportion (89%) of small, single layered follicles. Active caspase-3 was localized to both the oocyte and granulosa cells of follicles that were considered to be undergoing atresia, but was not localized to the granulosa cells of any small, single layered follicles. The results of this study indicate that, in small follicles, granulosa cell proliferation occurs independently of the size of follicles and the number of constituent cells, and that follicles of this type may be inherently less susceptible to the normal physiological factors that induce atresia.
Expression of caspase-2, -3, -8 and -9 proteins and enzyme activity in the corpus luteum of the rat at different stages during the natural estrous cycle. Peluffo MC et al. Apoptosis is associated with the regression of the corpus luteum (CL) in many species. Since caspases play a central role in apoptosis, we studied several initiators (-2, -8, and -9) and the main effector (-3) caspase in the CL during the estrous cycle of the rat. Two different populations of CL (old and new) were identified on ovaries at estrus and diestrus II (DII). Diminished (P < 0.05) luteal progesterone content and P450scc levels suggested that functional luteolysis occurred between the new CL at DII and old CL at estrus, whereas the decline (P < 0.05) in luteal weight indicated that structural regression was occurring between old CL at estrus to DII. Immunostaining for caspase-2 in luteal and endothelial cells appeared to increase as the luteal phase progressed, peaking at DII in the old CL. However, caspase-8 and -9 immunostaining showed little change with a slight increase at estrus in the old population. Notably, caspase-3 staining appeared to peak at DII in the new CL. Enzyme activity of caspase-9 increased (P < 0.05) in the new CL at DII, followed by that of caspase-2 and -3 in old CL at estrus. Caspase-8 activity did not change at any stage. The number of apoptotic cells increased at DII in the old CL. These results suggest an important role for this protease family during early events of luteolysis in the rat estrous cycle.
Phenotypes
Mutations
1 mutations
Species: mouse
Mutation name: CPP32-deficiency
type: null mutation fertility: unknown Comment: After finding high sequence homology of ced-3, a cell death gene in C. elegans, and CPP32, Kuida et al.(1996) generated CPP32-deficient mice by homologous recombination. These mice, born at a frequency lower than expected by mendelian genetics, were smaller than their littermates and died at 1-3 weeks of age. Although their thymocytes retained normal susceptibility to various apoptotic stimuli, brain development in CPP32-deficient mice was profoundly affected, and discernible by embryonic day 12, resulting in a variety of hyperplasias and disorganized cell deployment. These supernumerary cells were postmitotic and terminally differentiated by the postnatal stage. Pyknotic clusters at sites of major morphogenetic change during normal brain development were not observed in the mutant embryos, indicating decreased apoptosis in the absence of CPP32. Thus CPP32 was shown to play a critical role during morphogenetic cell death in the mammalian brain.