Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program

Nakahara, Kenji; Saito, Hidekazu; Saito, Toki; Ito, Mariko; Ohta, Nobuhiko; Sakai, Nobuyuki; Tezuka, Naohiro; Hiroi, Masahiko; Watanabe, Hiroshi

doi:10.1016/s0015-0282(97)81915-2

Cited by 117 publications

(96 citation statements)

References 25 publications

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“…First, the critical impact of apoptosis on the outcome of fertilization and oocyte development. In this regard, the inability to conceive is associated with a higher rate of apoptosis in their granulosa-lutein cells (32)(33)(34). Second, our study proposes the combined morphology and TUNEL assay as useful and simple strategy for the detection of apoptosis ovarian tissue destined for transplantation.…”

Section: Discussionmentioning

confidence: 88%

Histological evaluation and in situ localization of apoptosis in fresh and cryopreserved ovarian tissue

Hussein

Jeremias

Sharma

et al. 2002

Fertility and Sterility

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Objective: To study the feasibility of using combined morphology and terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) for apoptosis detection and the impact of cryopreservation on this process. Materials and methods:We conducted this investigation using a porcine animal model. Bilateral oophorectomy was performed in eight sows. The ovarian tissues were divided into two parts; one part was immediately fixed while the other was cryopreserved. The cryopreserved specimens were subsequently thawed and then fixed. All the specimens were sectioned, fixed and stained with H&E. The ovarian follicles were counted, histologically categorized as healthy and atretic and evaluated for the presence of apoptosis. Then, in situ examination of apoptosis was performed using TUNEL assay. Results: In the cryopreserved tissues: 1) the count of the primordial follicle was significantly reduced as compared to freshly-fixed samples (4.9±5.3 vs. 7.2±5.4, p=0.03 respectively) and 2) the mean values of apoptosis were insignificantly higher when compared to the freshly-fixed group (p=0.74). Moreover: 1) apoptosis was found in the atretic, but not in the healthy follicles (primordial, primary and secondary); 2) the nuclei of the granulosa cells, but not those of theca or stromal cells, were TUNEL positive; 3) some cells with histological features of necrosis and apoptosis were TUNEL negative, and 6) The distribution of apoptosis was not different between cryopreserved tissue and freshly fixed tissue. Conclusions: the presence of apoptosis in the atretic follicles may suggest its involvement in follicular atresia; and 2) combined histology and TUNEL assay may be a useful method for detection of apoptosis. Key words: Apoptosis/ovarian cryopreservation/Morphology/TUNEL assay.In 1885, the first observation of apoptosis in the ovary was made following morphological analysis of granulosa cells in the rabbit ovary (1). This form of cell death is involved in the ovarian homeostasis (2) and its regulation depends on a balance between (3,4). Morphologically, apoptosis is characterized by specific changes, such as cytoplasmic vacuolization, chromatin condensation and formation of apoptotic bodies (5, 6). Biochemically, apoptosis is characterized by DNA fragmentation into oligonucleosome-sized fragments (6). This DNA fragmentation has been examined in the pig ovarian tissues; using DNA fluorescence flowcytometry (7) and autoradiography (8-10). However, these methods have the following limitations 1) they require large amounts of DNA and 2) they do not allow identification of apoptotic changes in specific tissue compartments or at the single cell level. 163Although morphology provided the original criteria for apoptosis and therefore remains as a reliable indicator, detection of early apoptotic changes may be beyond the scope of histological examination. Alternatively, it is well known that apoptosis may occur without evidence of DNA fragmentation (11) and therefore its detection becomes beyond the scope...

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Section: Discussionmentioning

confidence: 88%

Histological evaluation and in situ localization of apoptosis in fresh and cryopreserved ovarian tissue

Hussein

Jeremias

Sharma

et al. 2002

Fertility and Sterility

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“…The incidence of apoptotic granulosa cells in a cumulus mass containing a subsequently fertilized oocyte was significantly lower than that in a cumulus mass with an unfertilized oocyte (9). Oxidative stress in granulose cells lowered fertilization rates and subsequently led to a decrease in the quality of the embryo (10).…”

Section: Discussionmentioning

confidence: 90%

“…We recently reported that it was possible to directly evaluate oocytes or embryos by measuring or monitoring intraovarian changes; the incidence of apoptotic granulosa cells (9), oxidative stress in granulosa cells (10), alternations in the cell cycle of granulosa cells (11), and concentrations of hyaluronan in follicular fluid (12). These intraovarian changes in the periovulatory phase might affect intraovarian blood flow and arterial resistance.…”

Section: Introductionmentioning

confidence: 99%

A Reduction in Intraovarian Arterial Blood Flow Resistance After Ovulation is Necessary to Achieve Pregnancy in Natural Cycle

Nakagawa

Ozawa

Takamatsu

et al. 2005

J Assist Reprod Genet

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Purpose : Color Doppler imaging permits the accurate localization of vessels and highfrequency pulsed Doppler ultrasonography has improved the resolution of flow velocity waveforms. In this study, intraovarian arterial blood flow before and after follicle rupture in the natural cycle was examined using transvaginal color flow Doppler imaging and changes in intraovarian arterial resistance in relation to the outcome of infertility treatment was analyzed. Methods : In a prospective study, 227 spontaneous cycles in 118 infertile patients who were undergoing infertility treatment at the division of Reproductive Medicine in our center were recruited in this study. The impedance to flow in intraovarian vessels was measured by means of transvaginal color flow Doppler imaging during the periovulatory period in the natural cycle of all patients. The pulsatility index (PI) of intraovarian arterial blood flow and pregnancy rate was evaluated. Results : On the basis of PI values before and after follicular rupture, 227 cycles were classified into severely decreased (113 cycles) and not-severely decreased groups (114 cycles). The pregnancy rate per cycle in the severely decreased group was 18.6% (21/113), significantly higher than that in the not-severely decreased group (7/114; 6.1%, p = 0.004). The miscarriage rate was similar in the two groups. Conclusions : A reduction in intraovarian blood vessel resistance is necessary to achieve pregnancy in a natural cycle.

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“…In humans, abnormally fertilized oocytes, as well as embryos arrested to grow in culture, demonstrate a high rate of mtDNA deletions, indicating the crucial role that mitochondria play in oocyte function and early embryo development [1,21]. Recent studies have shown a relationship between GC apoptosis and ART outcomes, but the data are still controversial [22].…”

Section: Discussionmentioning

confidence: 99%

Difference in mitochondrial gene expression in granulosa cells between recombinant FSH and hMG cycles under in vitro fertilization and transfer

Saito

Yamashita

Higuchi

et al. 2013

Reprod Medicine & Biology

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Purpose Examination of the mitochondrial mRNA expression in granulosa cells from an unspecified population of infertile patients to evaluate whether recombinant follicle stimulating hormone (recFSH) is more effective in producing higher quality embryo rates compared with human menopausal gonadotropin (hMG). Method Thirty-nine patients who underwent the in vitro fertilization and embryo transfer program were retrospectively examined. Patients were administered recFSH (n = 18) or hMG (n = 20) in a long protocol where GnRH agonist was used. Granulosa cells were obtained during oocyte retrieval and examined for mitochondria mRNA expression ratio against GAPDH. Expressions of mitochondria mRNA were evaluated by real-time PCR analysis. Results The high-quality embryo rate in the hMG cycle was higher than in the recFSH cycle, and the total dose of hMG showed a positive correlation with the expression level of mitochondrial genes in granulosa cells. Moreover, mitochondria mRNA expression was higher in the hMG cycle than in the recFSH cycle. Conclusions Compared with recFSH, hMG induces a higher mitochondrial gene expression ratio in granulosa cells at the time of oocyte retrieval and, therefore, may lead to higher quality embryo rates.

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Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program

Cited by 117 publications

References 25 publications

Histological evaluation and in situ localization of apoptosis in fresh and cryopreserved ovarian tissue

Histological evaluation and in situ localization of apoptosis in fresh and cryopreserved ovarian tissue

A Reduction in Intraovarian Arterial Blood Flow Resistance After Ovulation is Necessary to Achieve Pregnancy in Natural Cycle

Difference in mitochondrial gene expression in granulosa cells between recombinant FSH and hMG cycles under in vitro fertilization and transfer

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