scholarly journals Bovine oocytes in secondary follicles grow and acquire meiotic competence in severe combined immunodeficient mice

Zygote ◽  
2003 ◽  
Vol 11 (2) ◽  
pp. 139-149 ◽  
Author(s):  
Shoichiro Senbon ◽  
Atsushi Ota ◽  
Masao Tachibana ◽  
Takashi Miyano
Keyword(s):  
Scid Mice ◽  
Immunoblotting Analysis ◽  
Bovine Albumin ◽  
Immunodeficient Mice ◽  
Antral Follicles ◽  
Mean Diameter ◽  
The Mean ◽  
The One ◽  
Bovine Oocytes ◽  
Meiotic Competence

Cortical tissues containing only primordial and primary follicles, or secondary follicles 140-190 μm in diameter, were collected from bovine ovaries and xenografted under the kidney capsules of female severe combined immunodeficient (SCID) mice. Histological examination revealed that all grafts were well vascularised and contained surviving follicles at 4 or 6 weeks after grafting. Primordial and primary follicles survived but did not develop beyond the one-layer stage. Secondary follicles, on the other hand, had formed antra at 4 weeks after grafting. The mean diameter of secondary follicles, which was 165.2 ± 17.0 μm (n = 42) before grafting, had developed to 442.9 ± 77.9 μm (n = 37) and 592.9 ± 116.0 μm (n = 45) in diameter at 4 and 6 weeks after grafting, respectively. The mean diameter of oocytes, which was 55.1 ± 4.9 μm (n = 42) before grafting, also increased significantly (4 weeks: 105.6 ± 6.3 μm; 6 weeks: 122.2 ± 2.6 μm; p < 0.05). Oocytes were recovered from follicles that had developed to more than 400 μm in diameter after 6 weeks, and were subjected to subsequent mature culture. Of these oocytes, 34% (11/32) resumed meiosis and 6% (2/32) matured to the second metaphase. Follicular fluid in bovine antral follicles developed in SCID mice had the 69 kDa protein, which was detected by anti-mouse albumin antibody but not by anti-bovine albumin antibody in immunoblotting analysis. These results demonstrated that bovine secondary follicles develop to the antral stage in SCID mice, and that the oocytes in the follicles acquire the meiotic competence.

scholarly journals Fertilization and development of bovine oocytes grown in female SCID mice

Zygote ◽  
2005 ◽  
Vol 13 (4) ◽  
pp. 309-315 ◽  
Author(s):  
Shoichiro Senbon ◽  
Kyosuke Ishii ◽  
Yoshiyuki Fukumi ◽  
Takashi Miyano
Keyword(s):  
Chorionic Gonadotropin ◽  
Blastocyst Stage ◽  
Scid Mice ◽  
The Other ◽  
Cell Stage ◽  
Mean Diameter ◽  
Bovine Spermatozoa ◽  
The Mean ◽  
Equine Chorionic Gonadotropin ◽  
Bovine Oocytes

We previously reported that xenografted bovine secondary follicles developed to the antral stage in severe combined immunodeficient (SCID) mice. In the present study, bovine secondary follicles 100–240 μm in diameter were xenografted under the kidney capsules of female SCID mice for 6 and 8 weeks, and we examined the oocytes' fertilization and developmental abilities. Bovine follicles developed with prolongation of grafting and became significantly larger than those before grafting. Injection of equine chorionic gonadotropin (eCG) into host mice made some surviving follicles develop larger than the other follicles. Furthermore, bovine oocytes grew in the follicles, and the mean diameter of the oocytes was 100 μm or more at 6 and 8 weeks of transplantation. Bovine oocytes that had grown in eCG-stimulated SCID mice 8 weeks after grafting were subjected to maturation culture. Some of the oocytes that had grown to 110 μm or more matured to the second metaphase (7% of oocytes 110–119 μm and 44% of those >120 μm). When the oocytes were inseminated with bovine spermatozoa, 15% (6/39) formed a female and a male pronucleus, and 2 days after insemination 24% (18/75) of oocytes cleaved and 2% (2/75) developed to the 5- to 8-cell stage. However, no embryo reached the blastocyst stage. These results indicate that bovine oocytes grown in SCID mice could be fertilized but acquired insufficient competence for embryonic development in the present conditions.

Effect of androstenedione on the growth and meiotic competence of bovine oocytes from early antral follicles

Zygote ◽  
2011 ◽  
Vol 20 (4) ◽  
pp. 407-415 ◽  
Author(s):  
Hiroaki Taketsuru ◽  
Yuji Hirao ◽  
Naoki Takenouchi ◽  
Kosuke Iga ◽  
Takashi Miyano
Keyword(s):  
Granulosa Cell ◽  
Low Dose ◽  
Cell Complexes ◽  
Antral Follicles ◽  
Culture Substrate ◽  
17Β Estradiol ◽  
The Mean ◽  
Bovine Oocytes ◽  
Meiotic Competence

SummaryMedium that contains 17β-estradiol has been reported to support in vitro growth of bovine oocytes, isolated from early antral follicles, until the final stage. The aim of this study was to determine the effects of androstenedione in medium on such growing bovine oocytes. Oocyte–granulosa cell complexes were collected from early antral follicles and cultured for 14 days in medium supplemented with 17β-estradiol (0, 10 and 100 ng/ml) or androstenedione (0, 10 and 100 ng/ml). The mean diameter of oocytes measured after seeding on the culture substrate was 96.9 μm (n = 191). Either steroid was necessary for maintainance of the organization of oocyte–granulosa cell complexes over the 14-day culture period. In the 17β-estradiol- or the androstenedione-supplemented medium about 80% or 65%, respectively, of viable oocytes were recovered. In both groups the increase in oocyte size was significant after 14 days. The in vitro grown oocytes were cultured for a further 22–24 h for oocyte maturation; 13% and 30% of oocytes grown in the 10 and 100 ng/ml 17β-estradiol-supplemented medium reached metaphase II, respectively; more than 64% of oocytes grown in the androstenedione-supplemented medium matured to metaphase II. These results show that androstenedione, as 17β-estradiol, can maintain the viability of bovine oocyte–granulosa cell complexes and support the growth of oocytes, and that androstenedione promotes the acquisition of oocyte meiotic competence efficiently at a low dose.

scholarly journals Association between p34cdc2 levels and meiotic arrest in pig oocytes during early growth

Zygote ◽  
1995 ◽  
Vol 3 (4) ◽  
pp. 325-332 ◽  
Author(s):  
Yuji Hirao ◽  
Youki Tsuji ◽  
Takashi Miyano ◽  
Akira Okano ◽  
Masashi Miyake ◽  
...  
Keyword(s):  
Germinal Vesicle ◽  
Early Growth ◽  
Catalytic Subunit ◽  
The Other ◽  
Antral Follicles ◽  
Stages Of Growth ◽  
Mean Diameter ◽  
M Phase ◽  
Chromatin Configuration ◽  
Meiotic Competence

SummaryThe molecules involved in determining meiotic competence were determined in porcine oocytes isolated from preantral and antral follicles of different sizes. Oocytes isolated from preantral follicles had a mean diameter of 78 μm, contained diffuse filamentous chromatin in the germinal vesicle and were incapable of progressing from the G2 to the M phase of the cycle even after 72 h in culture. Oocytes from early antral follicles had a mean diameter of 105 μm, showed a filamentous chromatin configuration and about half resumed meiosis but arrested at metaphase I (MI) when cultured. Oocytes from mid-antral (3–4 mm) and large antral follicles (5–6 mm) had mean oocyte diameters of 115 and 119 μm respectively, contained condensed chromatin around the nucleolus and progressed to metaphase II (MII) in 48% and 93% of instances respectively. Analysis of p34cdc2, the catalytic subunit of maturation promoting factor (MPF), by immunoblotting indicates that the inability of small (78 μm) oocytes to resume meiosis is due, at least in part, to inadequate levels of the catalytic subunit of MPF. On the other hand, the inability of intermediate-sized (105 μm) oocytes from antral follicles to complete the first meiotic division by progressing beyond MI appears not to be limited by levels of p34cdc2, which are maximal by this stage. We postulate that an inadequacy of molecules other than p34cdc2 limits progression of MI to MII; the acquisition of these molecules during the final stages of growth may be correlated with the formation of the perinucleolar chromatin rim in the germinal vesicle.

scholarly journals DoE-It-Yourself: A Case Study for Implementing Design of Experiments into Nanoparticle Synthesis

2019 ◽  
Author(s):  
Niamh Mac Fhionnlaoich ◽  
Ye Yang ◽  
Runzhang Qi ◽  
Federico Galvanin ◽  
Stefan Guldin
Keyword(s):  
Gold Nanoparticles ◽  
Design Of Experiments ◽  
Marked Reduction ◽  
Nanoparticle Synthesis ◽  
Amount Of Information ◽  
Mean Diameter ◽  
The Mean ◽  
The One

Predictable and repeatable outcome is a major issue in nanoparticle synthesis. Traditionally, chemists rely on the one-factor-at-a-time method to investigate and optimise synthetic processes; however, this method is inefficient and often misleading. Design of Experiments (DoE), in contrast, can provide a greater amount of information in fewer experiments and lends itself to producing more reproducible results. Nevertheless, DoE techniques are only used by a relatively low number of practitioners in nanoparticle research. Here, we provide a case study on the synthesis of oleylamine-capped gold nanoparticles (AuNPs). Through the use of DoE, we achieved a marked reduction in dispersity and developed a model to carefully control the mean diameter of the nanoparticle populations.

Fertilization of bovine oocytes grown in vitro

1997 ◽  
Vol 9 (8) ◽  
pp. 781 ◽  
Author(s):  
Shigeru Osaki ◽  
Kenji Matsumura ◽  
Ken Yamamoto ◽  
Takashi Miyano ◽  
Masashi Miyake ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Calf Serum ◽  
Collagen Gels ◽  
Normal Morphology ◽  
Bovine Spermatozoa ◽  
The Mean ◽  
First Time ◽  
Bovine Oocytes ◽  
Meiotic Competence

Early antral follicles 0·5–0·7 mm in diameter were dissected from bovine ovaries, and oocyte–cumulus complexes with pieces of parietal granulosa (OCCGs) were then collected from the follicles. The OCCGs containing oocytes of 90–99 m diameter (94·7 ± 2·8 µm, n = 196) were selected and embedded in collagen gels and cultured for 14 days in TCM199 containing 10% fetal calf serum and 4 mM hypoxanthine. From cultured OCCGs, 144 surviving oocytes were recovered, of which 53 were granulosa cell-enclosed oocytes and 91 were denuded oocytes. The mean diameter of the surviving oocytes was 114·2 ± 8·4 µm, significantly larger than that measured before culture (P < 0·05). The granulosa cell-enclosed oocytes and denuded oocytes were further cultured for maturation for 24 h. After culture, 72% (38/53) of the granulosa cell-enclosed oocytes and 59% (54/91) of the denuded oocytes showed normal morphology. These oocytes were then inseminated with bovine spermatozoa. After 29 h of insemination, all of the denuded oocytes had degenerated, while 32% (12/38) of the granulosa cell-enclosed oocytes showed normal morphology. Of the 12 oocytes, 5 were penetrated by spermatozoa, and 2 formed both male and female pronuclei. These results demonstrate for the first time that bovine oocytes grown in vitro acquire meiotic competence and can be penetrated by spermatozoa.

Bovine oocytes from early antral follicles grow to meiotic competence in vitro: Effect of FSH and hypoxanthine

1997 ◽  
Vol 48 (5) ◽  
pp. 743-755 ◽  
Author(s):  
M. Harada ◽  
T. Miyano ◽  
K. Matsumura ◽  
S. Osaki ◽  
M. Miyake ◽  
...  
Keyword(s):  
Antral Follicles ◽  
Vitro Effect ◽  
Bovine Oocytes ◽  
Meiotic Competence

scholarly journals DoE-It-Yourself: A Case Study for Implementing Design of Experiments into Nanoparticle Synthesis

2019 ◽  
Author(s):  
Niamh Mac Fhionnlaoich ◽  
Ye Yang ◽  
Runzhang Qi ◽  
Federico Galvanin ◽  
Stefan Guldin
Keyword(s):  
Gold Nanoparticles ◽  
Design Of Experiments ◽  
Marked Reduction ◽  
Nanoparticle Synthesis ◽  
Amount Of Information ◽  
Mean Diameter ◽  
The Mean ◽  
The One

Predictable and repeatable outcome is a major issue in nanoparticle synthesis. Traditionally, chemists rely on the one-factor-at-a-time method to investigate and optimise synthetic processes; however, this method is inefficient and often misleading. Design of Experiments (DoE), in contrast, can provide a greater amount of information in fewer experiments and lends itself to producing more reproducible results. Nevertheless, DoE techniques are only used by a relatively low number of practitioners in nanoparticle research. Here, we provide a case study on the synthesis of oleylamine-capped gold nanoparticles (AuNPs). Through the use of DoE, we achieved a marked reduction in dispersity and developed a model to carefully control the mean diameter of the nanoparticle populations.

scholarly journals 225GROWTH OF OOCYTES IN PIG PRIMORDIAL FOLLICLES XENOTRANSPLANTED INTO SCID MICE

2004 ◽  
Vol 16 (2) ◽  
pp. 233 ◽  
Author(s):  
M. Moniruzzaman ◽  
S. Senbon ◽  
T. Miyano
Keyword(s):  
Histological Examination ◽  
Scid Mice ◽  
The Other ◽  
Oocyte Growth ◽  
Growth Property ◽  
Primordial Follicles ◽  
Mean Diameter ◽  
The Mean ◽  
Newborn Pigs ◽  
Cortical Slices

The mammalian ovary is endowed with a large number of primordial follicles that contain small oocytes. A limited number of these oocytes initiate growth, whereas others are either degenerate or remain as completely resting oocytes throughout the reproductive life of the female. The mechanism for the initiation of oocyte growth is not understood well. Small oocytes in primordial follicles of newborn rodents start to grow in the cultured ovary or ovarian tissue. For domestic animals, however, culture systems for mouse oocytes have not been valid. Xenotransplantation of ovarian tissues to immunodeficient mice can be a substitute for an effective culture system for small oocytes. Indeed, recent reports reveal the growth of small oocytes of fetal and newborn animals in xenografts (Hosoe et al., 2001;; Kaneko et al., 2003 Biol. Reprod. 53, 931–939). This experiment was conducted to study the growth of oocytes in primordial follicles of adult pig (6-month-old) in comparison to those of newborn pigs (10-day-old) in xenografts. The effect of the sex of host mice on oocyte growth in xenografts was also examined. Cortical slices containing only primordial follicles were collected under a dissection microscope from the ovaries of 6-month-old gilts (n=8) and 10-day-old piglets (n=6). Size of the slices was about 2mm×1mm×0.5mm. Each slice was cut into 2 pieces;; one was fixed for histological examination and the other was transplanted. For transplantation, 6- to 8-week-old male and female SCID (severe combined immune deficiency) mice were anesthetized, their left kidneys were exteriorized, and cortical slices were inserted under the kidney capsules. After 2 months, the grafts were recovered and processed for histological examination. Histological examination confirmed that the cortical slices contained only primordial follicles before transplantation. After transplantation, 47% (171/364) of the primordial follicles of adult pig ovaries survived in the xenografts but none of those developed into primary follicles or beyond. The mean diameter of the oocytes after transplantation was 32.1±0.7μm (n=171) which was similar to that of the oocytes before transplantation (30.7±0.8μm, n=364). On the other hand, in the xenografts of newborn pig ovaries, 13.5, 9.7 and 0.3% of 1122 follicles developed to the primary, secondary and antral stages, respectively, in male SCID mice. In the female SCID mice, there were no antral follicles but the distribution of primordial, primary and secondary follicles was 84.5, 9.4 and 6.1%, respectively, among a total of 1094 follicles. The mean diameter of the secondary follicles in the xenografts in the male SCID mice was 263.3±92.0μm (n=109) which was significantly higher than that of the secondary follicles in the female SCID mice (189.3±44.2μm, n=98) (P&lt;0.05, t-test). The results show that primordial follicles of 6-month-old pig ovaries survive but do not develop in xenografts, whereas newborn pig primordial follicles develop to the antral stage. This suggests that the growth property of the oocytes in primordial follicles in adult pig ovaries is different from that in newborn pigs.

Evaluation of Road Utilisation as Space For Holding Social Ceremonies in Indigenous Residential Area of Ogbomoso, Nigeria

2018 ◽  
Vol 1 (March 2018) ◽  
Author(s):  
S.A Okanlawon ◽  
O.O Odunjo ◽  
S.A Olaniyan
Keyword(s):  
Transport Infrastructure ◽  
Noisy Environment ◽  
Policy Makers ◽  
The Core ◽  
Local Roads ◽  
Strict Enforcement ◽  
The Mean ◽  
The One ◽  
The City ◽  
The Town

This study examined Residents’ evaluation of turning transport infrastructure (road) to spaces for holding social ceremonies in the indigenous residential zone of Ogbomoso, Oyo State, Nigeria. Upon stratifying the city into the three identifiable zones, the core, otherwise known as the indigenous residential zone was isolated for study. Of the twenty (20) political wards in the two local government areas of the town, fifteen (15) wards that were located in the indigenous zone constituted the study area. Respondents were selected along one out of every three (33.3%) of the Trunk — C (local) roads being the one mostly used for the purpose in the study area. The respondents were the residents, commercial motorists, commercial motorcyclists, and celebrants. Six hundred and forty-two (642) copies of questionnaire were administered and harvested on the spot. The Mean Analysis generated from the respondents’ rating of twelve perceived hazards listed in the questionnaire were then used to determine respondents’ most highly rated perceived consequences of the practice. These were noisy environment, Blockage of drainage by waste, and Endangering the life of the sick on the way to hospital; the most highly rated reasons why the practice came into being; and level of acceptability of the practice which was found to be very unacceptable in the study area. Policy makers should therefore focus their attention on strict enforcement of the law prohibiting the practice in order to ensure more cordial relationship among the citizenry, seeing citizens’ unacceptability of the practice in the study area.

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