Dynamic changes in mitochondrial 3D structure during folliculogenesis and luteal formation in the goat large luteal cell lineage

AbstractIn mammalian ovaries, mitochondria are integral sites of energy production and steroidogenesis. While shifts in cellular activities and steroidogenesis are well characterized during the differentiation of large luteal cells in folliculogenesis and luteal formation, mitochondrial dynamics during this process have not been previously evaluated. In this study, we collected ovaries containing primordial follicles, mature follicles, corpus hemorrhagicum, or corpus luteum from goats at specific times in the estrous cycle. Enzyme histochemistry, ultrastructural observations, and 3D structural analysis of serial sections of mitochondria revealed that branched mitochondrial networks were predominant in follicles, while spherical and tubular mitochondria were typical in large luteal cells. Furthermore, the average mitochondrial diameter and volume increased from folliculogenesis to luteal formation. In primordial follicles, the signals of cytochrome c oxidase and ATP synthase were undetectable in most cells, and the large luteal cells from the corpus hemorrhagicum also showed low enzyme signals and content when compared with granulosa cells in mature follicles or large luteal cells from the corpus luteum. Our findings suggest that the mitochondrial enlargement could be an event during folliculogenesis and luteal formation, while the modulation of mitochondrial morphology and respiratory enzyme expressions may be related to tissue remodeling during luteal formation.

Download Full-text

HIF-1α Activation Promotes Luteolysis by Enhancing ROS Levels in the Corpus Luteum of Pseudopregnant Rats

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/1764929 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Zonghao Tang ◽

Jiajie Chen ◽

Zhenghong Zhang ◽

Jingjing Bi ◽

Renfeng Xu ◽

...

Keyword(s):

Oxidative Stress ◽

Corpus Luteum ◽

Cell Apoptosis ◽

In Vitro Study ◽

Luteal Cell ◽

Independent Manner ◽

Luteal Regression ◽

Luteal Cells

The increase of oxidative stress is one of the important characteristics of mammalian luteal regression. Previous investigations have revealed the essential role of reactive oxygen species (ROS) in luteal cell death during luteolysis, while it is unknown how ROS is regulated in this process. Considering the decrease of blood flow and increase of PGF2α during luteolysis, we hypothesized that the HIF-1α pathway may be involved in the regulation of ROS in the luteal cell of the late corpus luteum (CL). Here, by using a pseudopregnant rat model, we showed that the level of both HIF-1α and its downstream BNIP3 was increased during luteal regression. Consistently, we observed the increase of autophagy level during luteolysis, which is regulated in a Beclin1-independent manner. Comparing with early (Day 7 of pseudopregnancy) and middle CL (Day 14), the level of ROS was significantly increased in late CL, indicating the contribution of oxidative stress in luteolysis. Inhibition of HIF-1α by echinomycin (Ech), a potent HIF-1α inhibitor, ameliorated the upregulation of BNIP3 and NIX, as well as the induction of autophagy and the accumulation of ROS in luteal cells on Day 21 of pseudopregnancy. Morphologically, Ech treatment delayed the atrophy of the luteal structure at the late-luteal stage. An in vitro study indicated that inhibition of HIF-1α can also attenuate PGF2α-induced ROS and luteal cell apoptosis. Furthermore, the decrease of cell apoptosis can also be observed by ROS inhibition under PGF2α treatment. Taken together, our results indicated that HIF-1α signaling is involved in the regression of CL by modulating ROS production via orchestrating autophagy. Inhibition of HIF-1α could obviously hamper the apoptosis of luteal cells and the process of luteal regression.

Download Full-text

Control of Steroidogenesis in Small and Large Bovine Luteal. Cells

Australian Journal of Biological Sciences ◽

10.1071/bi9870331 ◽

1987 ◽

Vol 40 (3) ◽

pp. 331 ◽

Cited By ~ 45

Author(s):

William Hansel ◽

Hector W Alila ◽

Joseph P Dowd ◽

Xiangzhong Yang

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Corpus Luteum ◽

Luteal Cell ◽

Lipoxygenase Pathway ◽

Luteal Cells ◽

Kinase C ◽

Bovine Corpus Luteum ◽

Progesterone Synthesis

Evidence was cited to show that: (1) prostacyclin (PGI2) plays a luteotrophic role in the bovine corpus luteum and that products of the lipoxygenase pathway of arachidonic acid metabolism, especially 5-hydroxyeicosatetraenoic acid play luteolytic roles; (2) oxytocin of luteal cell origin plays a role in development, and possibly in regression, of the bovine corpus luteum; and (3) luteal cells arise from two sources; the characteristic small luteal cells at all stages of the o~strous cycle and pregnancy are of theca cell origin; the large cells are of granulosa cell origin early in the cycle, but a population of theca-derived large cells appears later in the cycle. Results of in vitro studies with total dispersed cells and essentially pure preparations of large and small luteal cells indicate that : (1) the recently described Ca2+ -polyphosphoinositol-protein kinase C second messenger system is involved in progesterone synthesis in the bovine corpus luteum; (2) activation of protein kinase C is stimulatory to progesterone synthesis in the small luteal cells; (3) activation of protein kinase C has no effect on progesterone synthesis in the large luteal cells; and (4) protein kinase C exerts its luteotrophic effect in total cell preparations, in part at least, by stimulating the production of prostacyclin. The protein kinase C system may cause down regulation of LH receptors in the large cells.

Download Full-text

Mitochondrial dynamics in yeast cell death and aging

Biochemical Society Transactions ◽

10.1042/bst0391520 ◽

2011 ◽

Vol 39 (5) ◽

pp. 1520-1526 ◽

Cited By ~ 33

Author(s):

Ralf J. Braun ◽

Benedikt Westermann

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Mitochondrial Dynamics ◽

Progressive Increase ◽

Mitochondrial Morphology ◽

Aging Research ◽

Yeast Saccharomyces Cerevisiae ◽

Cell Death Pathways ◽

Dependent Cell ◽

Mitochondrial Networks

Mitochondria play crucial roles in programmed cell death and aging. Different stimuli activate distinct mitochondrion-dependent cell death pathways, and aging is associated with a progressive increase in mitochondrial damage, culminating in oxidative stress and cellular dysfunction. Mitochondria are highly dynamic organelles that constantly fuse and divide, forming either interconnected mitochondrial networks or separated fragmented mitochondria. These processes are believed to provide a mitochondrial quality control system and enable an effective adaptation of the mitochondrial compartment to the metabolic needs of the cell. The baker's yeast, Saccharomyces cerevisiae, is an established model for programmed cell death and aging research. The present review summarizes how mitochondrial morphology is altered on induction of cell death or on aging and how this correlates with the induction of different cell death pathways in yeast. We highlight the roles of the components of the mitochondrial fusion and fission machinery that affect and regulate cell death and aging.

Download Full-text

Is the canine corpus luteum an insulin-sensitive tissue?

Journal of Endocrinology ◽

10.1530/joe-16-0173 ◽

2016 ◽

Vol 231 (3) ◽

pp. 223-233 ◽

Cited By ~ 2

Author(s):

Liza Margareth Medeiros de Carvalho Sousa ◽

Renata dos Santos Silva ◽

Vanessa Uemura da Fonseca ◽

Rafael Magdanelo Leandro ◽

Thiago Senna Di Vincenzo ◽

...

Keyword(s):

Glucose Uptake ◽

Corpus Luteum ◽

Molecular Mechanisms ◽

Glucose Transporter ◽

Hypoxia Inducible Factor ◽

Glucose Disposal ◽

Luteal Cell ◽

Luteal Function ◽

Luteal Cells ◽

Glut4 Expression

This study aimed to determine in the canine corpus luteum throughout the dioestrus (1) the influence of insulin on glucose uptake; (2) the regulation of genes potentially involved; and (3) the influence of hypoxia on glucose transporter expression and steroidogenesis, after treatment with cobalt chloride (CoCl2). Glucose uptake by luteal cells increased 2.7 folds (P < 0.05) in response to insulin; a phenomenon related to increased expression of glucose transporter (GLUT) 4 and phosphorylation of protein kinase B (AKT). The gene expression of insulin receptor and SLC2A4 (codifier of GLUT4) genes after insulin stimulation increased on day 20 post ovulation (p.o.) and declined on day 40 p.o. (P < 0.05). Regarding potentially involved molecular mechanisms, the nuclear factor kappa B gene RELA was upregulated on days 30/40 p.o., when SLC2A4 mRNA was low, and the interleukin 6 (IL6) gene was upregulated in the first half of dioestrus, when SLC2A4 mRNA was high. CoCl2 in luteal cell cultures increased the hypoxia-inducible factor HIF1A/HIF1A and the SLC2A4/GLUT4 expression, and decreased progesterone (P4) production and hydroxyl-delta-5-steroid dehydrogenase 3 beta (HSD3B) mRNA expression (P < 0.05). This study shows that the canine luteal cells are responsive to insulin, which stimulates glucose uptake in AKT/GLUT4-mediated pathway; that may be related to local activity of RELA and IL6. Besides, the study reveals that luteal cells under hypoxia activate HIF1A-modulating luteal function and insulin-stimulated glucose uptake. These data indicate that insulin regulates luteal cells’ glucose disposal, participating in the maintenance and functionality of the corpus luteum.

Download Full-text

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

Journal of Endocrinology ◽

10.1677/joe.0.0390163 ◽

1967 ◽

Vol 39 (2) ◽

pp. 163-171 ◽

Cited By ~ 8

Author(s):

A. S. EL-SHEIKH ◽

FRANÇOIS B. SAKLA ◽

SAFAA O. AMIN

Keyword(s):

Corpus Luteum ◽

Cell Volume ◽

Distribution System ◽

Oestrous Cycle ◽

Luteal Cell ◽

Corpora Lutea ◽

Luteal Cells ◽

Functional Changes ◽

Parallel Increase ◽

Luteal Tissue

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.

Download Full-text

Varied effects of doxorubicin (DOX) on the corpus luteum of C57BL/6 mice during early pregnancy

Biology of Reproduction ◽

10.1093/biolre/ioab180 ◽

2021 ◽

Author(s):

Christian Lee Andersen ◽

Haeyeun Byun ◽

Yuehuan Li ◽

Shuo Xiao ◽

Doris M Miller ◽

...

Keyword(s):

Endothelial Cells ◽

Corpus Luteum ◽

Early Pregnancy ◽

Lipid Droplets ◽

Luteal Cell ◽

Knowledge Gap ◽

Chemotherapeutic Drugs ◽

Luteal Cells ◽

Single Intraperitoneal Injection ◽

Premenopausal Patients

Abstract Certain chemotherapeutic drugs are toxic to ovarian follicles. The corpus luteum (CL) is normally developed from an ovulated follicle for producing progesterone (P4) to support early pregnancy. To fill in the knowledge gap about effects of chemotherapy on the CL, we tested the hypothesis that chemotherapy may target endothelial cells and/or luteal cells in the CL to impair CL function in P4 steroidogenesis using doxorubicin (DOX) as a representative chemotherapeutic drug in mice. In both mixed background mice and C57BL/6 mice, a single intraperitoneal injection of DOX (10 mg/kg) on 0.5 days post coitum (D0.5, post-ovulation) led to ~58% D3.5 mice with serum P4 levels lower than the serum P4 range in the PBS-treated control mice. Further studies in the C57BL/6 ovaries revealed that CLs from DOX-treated mice with low P4 levels had less defined luteal cords and disrupted collagen IV expression pattern, indicating disrupted capillary, accompanied with less differentiated luteal cells that had smaller cytoplasm and reduced StAR expression. DOX-treated ovaries had increased granulosa cell death in the growing follicles, reduced PCNA-positive endothelial cells in the CLs, enlarged lipid droplets and disrupted F-actin in the luteal cells. These novel data suggest that the proliferating endothelial cells in the developing CL may be the primary target of DOX to impair the vascular support for luteal cell differentiation and subsequently P4 steroidogenesis. This study fills in the knowledge gap about the toxic effects of chemotherapy on the CL and provides critical information for risk assessment of chemotherapy in premenopausal patients.

Download Full-text

Cell death induced by serum deprivation in luteal cells involves the intrinsic pathway of apoptosis

Reproduction ◽

10.1530/rep.1.00751 ◽

2006 ◽

Vol 131 (1) ◽

pp. 103-111 ◽

Cited By ~ 30

Author(s):

Alicia A Goyeneche ◽

Jacquelyn M Harmon ◽

Carlos M Telleria

Keyword(s):

Cell Death ◽

Corpus Luteum ◽

Simian Virus 40 ◽

Full Length ◽

Luteal Cell ◽

Serum Starvation ◽

Endocrine Gland ◽

Luteal Cells ◽

Defined Culture

The corpus luteum is a transient endocrine gland specializing in the production of progesterone. The regression of the corpus luteum involves an abrupt decline in its capacity for producing progesterone followed by its structural involution, which is associated with apoptosis of the luteal cells. An in vitro experimental approach is needed to study the molecular mechanisms underlying hormonal regulation of luteal cell death under defined experimental conditions. In this study, we investigated simian virus-40-transformed luteal cells to determine whether they can be driven to apoptosis and, if so, to define the intracellular pathway involved. Luteal cells were cultured in the presence or absence of fetal bovine serum for 24 or 48 h. Under serum starvation conditions, the luteal cells underwent growth arrest accompanied by cell death as evaluated by dye exclusion, and confirmed by two-color fluorescence cell viability/cytotoxicity assay. We next studied whether serum starvation-induced death of luteal cells occurred by apoptosis. Morphologic features of apoptosis were observed in cells stained with hematoxylin after being subjected to serum starvation for 48 h. The apoptotic nature was further confirmed by in situ 3′-end labeling and fragmentation of genomic DNA. Apoptosis of serum-deprived luteal cells was dependent upon caspase activation. Serum starvation induced cleavage of poly (ADP-ribose) polymerase (PARP), suggesting that caspase-3 had been activated under the stress of withdrawal of growth factors. This was confirmed by cleavage of full-length procaspase-3. Finally, the fact that serum starvation promoted the cleavage of full-length procaspase-9 and the decrease in the expression of endogenous Bid, a BH-3-only proapoptotic protein of the Bcl-2 family, indicates that the intrinsic (i.e., mitochondrial) pathway of apoptosis was activated. In summary, we have characterized an in vitro experimental model of luteal cell death that can be utilized to evaluate the role of hormones in apoptosis of luteal cells under defined culture conditions, and to study the mechanism of luteal regression.

Download Full-text

Regulation of mitochondrial morphology by APC/CCdh1-mediated control of Drp1 stability

Molecular Biology of the Cell ◽

10.1091/mbc.e10-07-0567 ◽

2011 ◽

Vol 22 (8) ◽

pp. 1207-1216 ◽

Cited By ~ 66

Author(s):

Sarah R. Horn ◽

Michael J. Thomenius ◽

Erika Segear Johnson ◽

Christopher D. Freel ◽

Judy Q. Wu ◽

...

Keyword(s):

Mitochondrial Dynamics ◽

Dynamic Balance ◽

Mitochondrial Morphology ◽

Anaphase Promoting Complex ◽

Cellular Division ◽

Mitochondrial Division ◽

Unequal Distribution ◽

Daughter Cells ◽

Ubiquitin Ligase Complex ◽

Mitochondrial Networks

Homeostatic maintenance of cellular mitochondria requires a dynamic balance between fission and fusion, and controlled changes in morphology are important for processes such as apoptosis and cellular division. Interphase mitochondria have been described as an interconnected network that fragments as cells enter mitosis, and this mitotic mitochondrial fragmentation is known to be regulated by the dynamin-related GTPase Drp1 (dynamin-related protein 1), a key component of the mitochondrial division machinery. Loss of Drp1 function and the subsequent failure of mitochondrial division during mitosis lead to incomplete cytokinesis and the unequal distribution of mitochondria into daughter cells. During mitotic exit and interphase, the mitochondrial network reforms. Here we demonstrate that changes in mitochondrial dynamics as cells exit mitosis are driven in part through ubiquitylation of Drp1, catalyzed by the APC/CCdh1 (anaphase-promoting complex/cyclosome and its coactivator Cdh1) E3 ubiquitin ligase complex. Importantly, inhibition of Cdh1-mediated Drp1 ubiquitylation and proteasomal degradation during interphase prevents the normal G1 phase regrowth of mitochondrial networks following cell division.

Download Full-text

Expression and regulation of secreted phosphoprotein 1 in the bovine corpus luteum and effects on T lymphocyte chemotaxis

Reproduction ◽

10.1530/rep-13-0190 ◽

2013 ◽

Vol 146 (6) ◽

pp. 527-537 ◽

Cited By ~ 7

Author(s):

Daniel H Poole ◽

Kalidou Ndiaye ◽

Joy L Pate

Keyword(s):

Corpus Luteum ◽

Estrous Cycle ◽

Cell Function ◽

Luteal Cell ◽

Cellular Interactions ◽

Western Blots ◽

Luteal Regression ◽

Luteal Cells ◽

Bovine Corpus Luteum ◽

Secreted Phosphoprotein 1

Secreted phosphoprotein 1 (SPP1) in the bovine corpus luteum (CL) regulates cell function during the transitional periods of luteinization and luteal regression. The objectives were to i) characterize SPP1 expression in the CL throughout the estrous cycle, ii) determine factors that regulate SPP1 expression in luteal cells, and iii) examine the role of SPP1 in lymphocyte chemotaxis, proliferation, and function.SPP1mRNA was greater in fully functional (d10) CL and late cycle (d18) CL compared with developing (d4) CL. Additionally,SPP1mRNA increased within 1 h and remained elevated 4 and 8 h following induction of luteolysis with prostaglandin (PG)F2α. Expression of the SPP1 receptor, β3integrin, was not different throughout the estrous cycle but decreased following induction of luteolysis. Expression ofCD44increased during the estrous cycle but did not change during luteal regression. In cultured luteal cells,SPP1mRNA was upregulated by PGF2αand/or tumor necrosis factor α. Western blots revealed the presence of both full-length SPP1 and multiple cleavage products in cultured luteal cells and luteal tissue. Depletion of endogenous SPP1 did not hinder luteal cell-induced lymphocyte proliferation or lymphocyte phenotype but did inhibit lymphocyte migration toward luteal cells. Based on these data, it is concluded that SPP1 is initially activated to establish and maintain cellular interactions between steroidogenic and nonsteroidogenic cells during the development of the CL. Upon induction of luteolysis, SPP1 serves as a signaling molecule to recruit or activate immune cells to facilitate luteal regression and tissue degradation.

Download Full-text

Deciphering the functional role of EGR1 in Prostaglandin F2 alpha induced luteal regression applying CRISPR in corpus luteum of buffalo

Biological Research ◽

10.1186/s40659-021-00333-7 ◽

2021 ◽

Vol 54 (1) ◽

Author(s):

Meeti Punetha ◽

Sai Kumar ◽

Avishek Paul ◽

Bosco Jose ◽

Jaya Bharati ◽

...

Keyword(s):

Mrna Expression ◽

Corpus Luteum ◽

Caspase 3 ◽

Cellular Proliferation ◽

Luteal Cell ◽

Wild Type ◽

Luteal Regression ◽

Luteal Cells ◽

Prostaglandin F2 Alpha

Abstract Background PGF2α is essential for the induction of the corpus luteum regression which in turn reduces progesterone production. Early growth response (EGR) proteins are Cys2-His2-type zinc-finger transcription factor that are strongly linked to cellular proliferation, survival and apoptosis. Rapid elevation of EGR1 was observed after luteolytic dose of PGF2α. EGR1 is involved in the transactivation of many genes, including TGFβ1, which plays an important role during luteal regression. Methods The current study was conducted in buffalo luteal cells with the aim to better understand the role of EGR1 in transactivation of TGFβ1 during PGF2α induced luteal regression. Luteal cells from mid stage corpus luteum of buffalo were cultured and treated with different doses of PGF2α for different time durations. Relative expression of mRNAs encoding for enzymes within the progesterone biosynthetic pathway (3βHSD, CYP11A1 and StAR); Caspase 3; AKT were analyzed to confirm the occurrence of luteolytic event. To determine if EGR1 is involved in the PGF2α induced luteal regression via induction of TGFβ1 expression, we knocked out the EGR1 gene by using CRISPR/Cas9. Result The present experiment determined whether EGR1 protein expression in luteal cells was responsive to PGF2α treatment. Quantification of EGR1 and TGFβ1 mRNA showed significant up regulation in luteal cells of buffalo at 12 h post PGF2α induction. In order to validate the role of PGF2α on stimulating the expression of TGFβ1 by an EGR1 dependent mechanism we knocked out EGR1. The EGR1 ablated luteal cells were stimulated with PGF2α and it was observed that EGR1 KO did not modulate the PGF2α induced expression of TGFβ1. In PGF2α treated EGR1 KO luteal cell, the mRNA expression of Caspase 3 was significantly increased compared to PGF2α treated wild type luteal cells maintained for 12 h. We also studied the influence of EGR1 on steroidogenesis. The EGR1 KO luteal cells with PGF2α treatment showed no substantial difference either in the progesterone concentration or in StAR mRNA expression with PGF2α-treated wild type luteal cells. Conclusion These results suggest that EGR1 signaling is not the only factor which plays a role in the regulation of PGF2α induced TGFβ1 signaling for luteolysis.

Download Full-text