206.Cell death of the theca interna during bovine ovarian follicular atresia

It is generally accepted that death of cells within the theca interna occurs late during ovarian follicular atresia. Histological classifications of atresia are usually based solely upon the morphology of the membrana granulosa. Atresia of bovine antral follicles less than 5 mm in diameter has been redefined as either antral or basal atresia depending on where in the membrana granulosa cell death is initiated. The aim of present study was to investigate changes within the theca interna during both antral and basal atresia. Bovine ovaries were collected and processed for light microscopy and immunohistochemistry. Each follicle less then 5 mm was classified as either healthy, antral atretic or basal atretic, with antral atresia being further classified either early-mid or late stage. Sections were labelled by TUNEL to identify dead cells combined with lectin from Bandeiraea simplificifolia to identify endothelial cells or with an antibody to cytochrome P450 cholesterol side-chain cleavage to identify steroidogenic cells. The numerical density of steroidogenic cells within the theca interna was significantly reduced (P < 0.001) in basal atretic follicles compared to healthy and antral atretic follicles. In both antral and basal atresia there was death of endothelial cells and steroidogenic cells. However cell death was greater in endothelial cells (P < 0.05) and steroidogenic cells (P < 0.001) of the theca interna of basal atretic follicles. There was no significant difference in the amount of cell death in the membrana granulosa between early-mid antral atresia and basal atresia while death of the membrana granulosa was significantly increased in late antral atresia compared to basal atresia (P < 0.01). Therefore we conclude that basal atresia is not a progression of antral atresia and that the theca interna can be susceptible to cell death early in atresia in basal atretic follicles.

Download Full-text

EFFECT OF ADRENOCORTICOTROPHIN ON INTRACELLULAR CHOLESTEROL TRANSPORT

Journal of Endocrinology ◽

10.1677/joe.0.0840179 ◽

1980 ◽

Vol 84 (2) ◽

pp. 179-188 ◽

Cited By ~ 32

Author(s):

MASAHISA NAKAMURA ◽

MASAAKI WATANUKI ◽

B. E. TILLEY ◽

P. F. HALL

Keyword(s):

Mitochondrial Membrane ◽

Cholesterol Content ◽

Tumour Cells ◽

Side Chain ◽

Adrenal Tumour ◽

Inner Mitochondrial Membrane ◽

Side Chain Cleavage ◽

Isolated Mitochondria ◽

Chain Cleavage ◽

Significant Difference

The rate of side-chain cleavage of cholesterol by mitochondria derived from mouse adrenal tumour cells was measured. Incubation of the cells in the presence of adrenocorticotrophin (ACTH) for periods of up to 1 h was without effect on the subsequent side-chain cleavage by mitochondria. However, if the cells were incubated in the presence of aminoglutethimide phosphate (0·76 mmol/l), addition of ACTH (final concentration 86 u./l) to the medium containing the cells increased the subsequent rate of side-chain cleavage by the isolated mitochondria. This response reached a maximum after incubation of cells with ACTH for 2 h and decayed when the isolated mitochondria were left at 0 °C, although a significant difference was still apparent after 120 min. Similar stimulation of mitochondrial side-chain cleavage by ACTH was observed when the reaction was inhibited by anaerobiosis instead of aminoglutethimide phosphate. Addition of dibutyryl cyclic AMP, at final concentrations greater than 10−5 mol/l, to cells during incubation with aminoglutethimide phosphate (0·76 mmol/l) also stimulated the conversion of cholesterol to pregnenolone by the mitochondria. Provided the adrenal tumour cells were incubated with aminoglutethimide, or anaerobically, the mean cholesterol content of the inner mitochondrial membrane was significantly higher (P < 0·01) when ACTH was included in the incubation medium than when it was not. It is concluded that ACTH increases the movement of cholesterol to the mitochondrial membrane which contains the side-chain cleavage enzyme system and that part of this cholesterol is used for the enhanced conversion of cholesterol to pregnenolone brought about by ACTH.

Download Full-text

Distribution of Cytochrome P450-side Chain Cleavage in the Theca Interna Layers of Bovine Small Antral and Cystic Follicles

Reproduction in Domestic Animals ◽

10.1046/j.1439-0531.2003.00456.x ◽

2003 ◽

Vol 38 (5) ◽

pp. 405-409 ◽

Cited By ~ 9

Author(s):

N Isobe ◽

T Nakao ◽

Y Yoshimura

Keyword(s):

Cytochrome P450 ◽

Side Chain ◽

Side Chain Cleavage ◽

Chain Cleavage ◽

Theca Interna

Download Full-text

Regulation of Cholesterol Side-Chain Cleavage and 17α-Hydroxylase/Lyase Activities in Proliferating Human Theca Interna Cells in Long Term Monolayer Culture*

Endocrinology ◽

10.1210/endo-125-4-1959 ◽

1989 ◽

Vol 125 (4) ◽

pp. 1959-1966 ◽

Cited By ~ 40

Author(s):

JAN M. McALLISTER ◽

JOHN F. P. KERIN ◽

JOHN M. TRANT ◽

RONALD W. ESTABROOK ◽

J. IAN. MASON ◽

...

Keyword(s):

Monolayer Culture ◽

Side Chain ◽

Side Chain Cleavage ◽

Chain Cleavage ◽

Theca Interna

Download Full-text

Ovarian Follicular Atresia of Ewes during Spring Puerperium

Veterinary Medicine International ◽

10.1155/2012/638928 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6

Author(s):

Radoslava Vlčková ◽

Drahomíra Sopková ◽

Ján Pošivák ◽

Igor Valocký

Keyword(s):

Follicular Atresia ◽

Follicular Cells ◽

Ovarian Surface ◽

Significant Difference ◽

Mean Diameter ◽

Low Levels ◽

Theca Interna ◽

Follicular Cysts

The distribution of healthy and atretic follicles on the ovarian surface of improved Valachian ewes 17, 24, and 32 days postpartum is reported in this study. The number of healthy follicles was higher on day 24 postpartum and their mean diameter tended to increase to day 32 (P<0.05) with the greatest diameter of 5 mm. 78–81% of atretic follicles ≥3 mm in diameter was observed where apoptosis began in the follicular cells situated at the follicular cavity. The early atretic follicles are characterized by the presence of mitotic pictures. In one ewe 24 days postpartum, small regressive follicular cysts were observed. Contracting atresia is characterized by thickening of the theca interna even to 190 μm. Progesterone and oestradiol-17βconcentrations were maintained at relatively low levels, but with no significant difference between the days postpartum.

Download Full-text

Thrombospondin-1 Expression Is Increased during Follicular Atresia in the Primate Ovary

Endocrinology ◽

10.1210/en.2007-0835 ◽

2007 ◽

Vol 149 (1) ◽

pp. 185-192 ◽

Cited By ~ 25

Author(s):

Fiona H. Thomas ◽

Helen Wilson ◽

Audrey Silvestri ◽

Hamish M. Fraser

Keyword(s):

Endothelial Cells ◽

Granulosa Cells ◽

Follicular Phase ◽

Follicular Atresia ◽

Gonadotropin Secretion ◽

Antral Follicles ◽

Cd36 Expression ◽

Mrna And Protein Expression ◽

Vegf Trap

Thrombospondin (TSP)-1 is an antiangiogenic extracellular matrix glycoprotein that modulates several aspects of cellular function. The aim of this study was to determine the pattern of TSP-1 mRNA and protein expression as well as expression of its receptor CD36 in the marmoset ovary and to investigate the effects of inhibition of gonadotropins or VEGF activity on TSP-1 and CD36 expression in vivo. GnRH antagonist or VEGF Trap, a soluble decoy receptor, was administered on d 0 of the follicular phase of the cycle, and ovaries were collected at the end of the follicular phase (d 10). TSP-1 mRNA and protein were present in granulosa cells of preantral and antral follicles, with the highest staining at the late secondary and tertiary stages. Moreover, expression of TSP-1 mRNA and protein was significantly increased in tertiary follicles undergoing atresia. CD36 protein was detected in granulosa cells of preantral and antral follicles as well as in endothelial cells of large vessels. Inhibition of gonadotropin secretion or VEGF activity had no effect on TSP-1 expression; however, expression of CD36 protein was inhibited by the VEGF Trap. In conclusion, TSP-1 may be involved in the cessation of angiogenesis in follicles undergoing atresia; alternatively, TSP-1 may act on granulosa and/or endothelial cells to promote follicular atresia in the ovary. Angiogenesis is likely to involve a balance between pro- and antiangiogenic factors. Our results suggest that loss of VEGF activity does not regulate TSP-1 expression directly but may influence TSP-1 activity via down-regulation of the CD36 receptor.

Download Full-text

222THE ROLE OF TUMOR NECROSIS FACTOR-RELATED APOPTOSIS INDUCING LIGAND DURING FOLLICULAR ATRESIA IN PIG OVARIES

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab222 ◽

2004 ◽

Vol 16 (2) ◽

pp. 232

Author(s):

N. Inoue ◽

N. Manabe ◽

M. Nakayama ◽

T. Matsui ◽

A. Maeda ◽

...

Keyword(s):

Cell Death ◽

Tumor Necrosis Factor ◽

Granulosa Cells ◽

Tumor Necrosis ◽

Mrna Levels ◽

Dependent Manner ◽

Follicular Atresia ◽

Rt Pcr ◽

Antral Follicles ◽

Necrosis Factor

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce cell death by binding to its receptors (DR4 and DR5). However, binding to DcR1 or DcR2 cannot induce apoptosis. DcRs compete with DRs. TRAIL has been reported to induce apoptosis in various tumor cells but not in normal cells. However, a recent study revealed that TRAIL induces apoptosis in normal hepatocytes of human but not in those of rat, mouse, or rhesus monkey, indicating that there are species-specific differences in TRAIL and receptor systems. In the present study, we demonstrated Immunohistochemical, Western immunoblotting, and reverse transcription-polymerase chain reaction analyses (RT-PCR) of TRAIL and DR4 in granulosa cells during follicular atresia in pig ovaries. For immunohistochemistry, pig ovaries obtained at a local slaughterhouse were fixed with 20% buffered formalin. For Western blotting and RT-PCR analysis, individual preovulatory antral follicles were dissected from the ovaries. Based on morphological and endocrinological criteria, the antral follicles were divided into three categories as follows: healthy, early stage of atresia, progressed stage of atresia. Significant increases were demonstrated in TRAIL protein and mRNA levels during atresia, but not in DR4 protein. Moreover in an in vitro apoptosis-inducing assay using cultured granulosa cells prepared from healthy follicles, we showed that more than 200ng/mL TRAIL could activate caspase-3 and induce apoptotic cell death in a dose-and time-dependent manner, but less than 100ng/mL of TRAIL could not induce apoptosis. When DcR1 was removed from the cell membrane of granulosa cells, a lower dose of TRAIL could induce apoptosis. The present findings suggested that the TRAIL can induce granulosa cell apoptosis, and that DcR1 blocks TRAIL-induced apoptosis in granulosa cells of healthy follicles in porcine ovaries.

Download Full-text