Structure and functions of the placenta in common minke (Balaenoptera acutorostrata), Bryde's (B. brydei) and sei (B. borealis) whales.

The structure and functions of placentas were examined in 3 species of rorqual whales, common minke (Balaenoptera acutorostrata), Bryde's (B. brydei) and sei (B. borealis) whales, with the aim of confirming the structural characteristics of the chorion, including the presence of the areolar part, and clarifying steroidogenic activities and fetomaternal interactions in the placentas of these whales.

Areolae of the placenta in the Antarctic minke whale (Balaenoptera bonaerensis).

In this study, we examined the existence and structure of areolae and the steroidogenesis of areolar trophoblast cells in the Antarctic minke whale placenta morphologically and immunohistochemically. Placentas were collected from the 15th, 16th and 18th Japanese Whale Research Program under Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. The opening and cavity of fetal areolae formed by taller columnar trophoblast cells (areolar trophoblast cells) with long microvilli and a bright cytoplasm, as compared with the trophoblast cells of the chorionic villi interdigitating with the endometrial crypts, were recognized in observations of serial sections.

Variation of cholesterol contents in porcine cumulus-oocyte complexes is a key factor in regulation of fertilizing capacity.

The aims were to explore substances affecting maturation of porcine oocytes and effects of cholesterol efflux by methyl-β-cyclodextrin (MBCD). Cumulus-oocyte complexes (COCs) were collected from ovaries with or without corpora lutea (CL). Ovarian cholesterol content was determined and histologic sections were prepared for immunostaining of lanosterol 14α-demethylase, a catalytic enzyme during cholesterogenesis.

Structure and steroidogenesis of the placenta in the Antarctic minke whale (Balaenoptera bonaerensis).

There are few reports describing the structure and function of the whale placenta with the advance of pregnancy. In this study, therefore, the placenta and nonpregnant uterus of the Antarctic minke whale were observed morphologically and immunohistochemically. Placentas and nonpregnant uteri were collected from the 15th, 16th and 18th Japanese Whale Research Programme with Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan.

A novel method for detection of chromosomal integrity in cryopreserved livestock spermatozoa using artificially fused mouse oocytes.

Purpose: The aim of the present study was to investigate the effect of mouse oocyte volume on the efficiency of chromosomal analysis in livestock spermatozoa.

Methods: Oocytes were injected with bull, ram, boar and dog sperm heads, and then fused with enucleated mouse oocytes.

Results: The increment of oocyte volume increased the rates of morphologically normal oocytes after sperm injection, which induced much higher rates of overall chromosome detection in bull, ram and dog spermatozoa.

Factors affecting the fertility of ewes after intrauterine insemination with frozen-thawed semen during the non-breeding season.

In this study, two successive field trials were conducted during the non-breeding season to investigate various factors affecting on fertility of Suffolk ewes after intrauterine insemination with frozen-thawed semen. In the first year (Experiment 1), three sperm numbers per insemination dose (0.25, 0.

Reproductive ability after uterine artery embolization in a sheep model: observation over 2 seasons.

Objective: To investigate the mid-term effect of uterine artery embolization (UAE) on fertility after bilateral UAE with either tris-acryl gelatin microspheres (TAGM) or gelatin particles (GP).

Materials And Methods: Fertility was compared in 6 ewes that underwent UAE with TAGM, 6 ewes that underwent UAE with GP, and 6 control ewes. All ewes were artificially inseminated or naturally bred for 2 consecutive breeding seasons after UAE.

Effects of seasonal changes on in vitro developmental competence of porcine oocytes.

The aim of this study was to investigate whether seasonal changes affected in vitro developmental competence of porcine oocytes. The relationship between atmospheric temperature and embryonic development of in vitro matured porcine oocytes following intracytoplasmic sperm injection was examined throughout the year. The blastocyst rate (31.

Fertilizability, developmental competence, and chromosomal integrity of oocytes microinjected with pre-treated spermatozoa in mice.

The aim of the present study was to investigate the safety of sperm pre-treatment during the ICSI procedure using a mouse model. Mouse spermatozoa were treated with methyl-beta-cyclodextrin, lysolecithin, Triton X-100, and dithiothreitol (DTT), and injected into mouse oocytes. The injected oocytes were monitored for chromosomal integrity and pre- and post-implantation development.

Fertility after uterine artery embolization: investigation using a sheep model.

Objective: To investigate the influence of uterine artery embolization (UAE) on fertility after bilateral UAE with either tris-acryl gelatin microspheres (TAGM) or gelatin particles (GP).

Materials And Methods: Six ewes that underwent UAE with TAGM, 6 ewes that underwent UAE with GP, and 6 control ewes were compared. After hormonal synchronization of the menstrual cycle, artificial insemination (AI) was performed.

Production of cloned sei whale (Balaenoptera borealis) embryos by interspecies somatic cell nuclear transfer using enucleated pig oocytes.

In this study, we examined the feasibility of using subzonal cell injection with electrofusion for interspecies somatic cell nuclear transfer (iSCNT) to produce sei whale embryos and to improve their developmental capacity by investigating the effect of osmolarity and macromolecules in the culture medium on the in vitro developmental capacity. Hybrid embryos produced by the electrofusion of fetal whale fibroblasts with enucleated porcine oocytes were cultured in modified porcine zygote medium-3 to examine the effects of osmolarity and fetal serum on their in vitro developmental capacity. More than 66% of the whale somatic cells successfully fused with the porcine oocytes following electrofusion.

Production of Sei whale (Balaenoptera borealis) cloned embryos by inter- and intra-species somatic cell nuclear transfer.

The objectives of this study were to choose an effective embryo reconstruction method and an effective post-activation agent for in vitro production of sei whale (Balaenoptera borealis) interspecies somatic cell nuclear transfer (iSCNT) embryos. Moreover, trichostatin A (TSA) treatment of whale iSCNT embryos was performed to improve the in vitro embryo development. In Experiment 1, the fusion rate was significantly higher (88.

Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes.

Purpose: The aim of the present study was to investigate the fertilizing capacity of fresh, frozen-thawed and freeze-dried canine spermatozoa.

Methods: After canine spermatozoa were injected into mouse oocytes, the rates of oocyte activation, male pronuclear formation and chromosomal aberrations were investigated.

Results: The rates of oocyte activation were comparable (90.

Effects of the presence and the numbers of corpora lutea in non-delivered and delivered pigs on in vitro oocyte maturation and embryonic development.

The aim of this study was to investigate the effects of the presence and the numbers of corpora lutea (CL) in porcine ovaries on in vitro oocyte maturation and embryonic development following intracytoplasmic sperm injection (ICSI). At oocyte collection, the ovaries of non-delivered and delivered pigs were classified into four groups by CL presence. The effect of the number of CL was also investigated following re-division of the non-delivered groups into four groups.

Effects of semen extenders and storage temperatures on characteristics of frozen-thawed Bryde's (Balaenoptera edeni) whale spermatozoa.

The present study investigated effects of three semen extenders and storage temperatures on post-thaw characteristics of Bryde's whale spermatozoa. Spermatozoa were collected from the vasa deferens of three mature Bryde's whales captured during the Japanese whale research in the north-west Pacific (May to August 2007) after death. The three semen extenders used for freezing were 1) a commercialized synthetic extender (AndroMed: AM), 2) Tris-based + 10% bovine serum albumin (BSA) and 3) Tris-based + egg yolk (EY).

Fertility after different artificial insemination methods using a synthetic semen extender in sheep.

The present study aimed to investigate the fertility of ewes artificially inseminated with three different methods using a synthetic semen extender, AndroMed. The three methods of artificial insemination (AI) were cervical AI with fresh-diluted or frozen-diluted semen at observed estrus, and an intrauterine AI with frozen-thawed semen. A total of 80 ewes were treated with a controlled internal drug release (CIDR) containing 0.

Fertility after artificial insemination using a soybean-based semen extender in sheep.

The present study aimed to compare the fertility of ewes intrauterinally inseminated with frozen-thawed semen using a soybean-based semen extender (AndroMed) with those of ewes intrauterinally inseminated with frozen-thawed semen using a Tris-based extender containing either egg yolk or BSA. Suffolk ewes (n=104) were treated with an intravaginal sponge containing 40 mg fluoroprogesterone acetate (FGA) for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin to induce estrus and ovulation during the non-breeding season (July, 2007). Intrauterine insemination was carried out 40-46 h after removal of the FGA sponge (n=90), regardless of the incidence of estrus.

Chemical properties of epidermal lipids, especially sphingolipids, of the Antarctic minke whale.

It is well known that sphingolipids specifically exist in the terrestrial mammal epidermis and correlate with skin barrier functions. However, the lipid properties of the marine mammal epidermis have not been examined in detail. We thus investigated the chemical composition of lipid components, especially sphingolipids, in the black epidermis (outer skin) of Antarctic minke whales (six mature and six immature specimens).

Follicle size-dependent changes in follicular fluid components and oocyte diameter in Antarctic minke whales (Balaenoptera bonaerensis).

The concentrations of various components of follicular fluid were compared among three groups of follicles (small, <5 mm; medium: 5-10 mm; large, >10 mm) with a control that consisted of the components of umbilical serum using seven pregnant Antarctic minke whales. Follicular oocytes recovered from the follicles were also used for measurement of oocyte diameter after removing the cumulus cells. The mean diameter of the ooplasm in the oocytes from the large follicles (143.

Attempt at intracytoplasmic sperm injection of in vitro matured oocytes in common minke whales (Balaenoptera acutorostrata) captured during the Kushiro Coast Survey.

The present study was conducted during the Kushiro Coast Survey in an attempt to produce common minke whale embryos. In Experiment 1, we attempted to determine the appropriate culture duration (30 or 40 h) for in vitro maturation (IVM) of immature oocytes using the Well of the Well method. In Experiment 2, and intracytoplasmic sperm injection (ICSI) was applied to matured oocytes from prepubertal and adult common minke whales after IVM culture (40 or 48 h), and then their embryonic development was assessed.

Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin.

The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal.

Chemical characterization of the oligosaccharides in Bryde's whale (Balaenoptera edeni) and Sei whale (Balaenoptera borealis lesson) milk.

Samples of milk from a Bryde's whale and a Sei whale contained 2.7 g/100 mL and 1.7 g/100 mL of hexose, respectively.

Changes of fructose concentrations in seminal plasma and glucose and testosterone concentrations in blood plasma in rams over the course of a year.

The present study was performed to examine seasonal changes in the fructose concentrations of seminal plasma and glucose and testosterone concentrations of blood plasma over the course of a year (from November 2004 to November 2005) using 5 Suffolk rams. Osmolality of the seminal plasma was also measured. The fructose concentrations in the seminal plasma increased as the breeding season approached, with the maximum in October (179.

In vitro culture conditions using chemically defined media for in vitro matured and intracytoplasmically inseminated porcine oocytes.

The present study investigated in vitro culture methods [droplet and Well of the Well (WOW)] using semi-defined and defined media [modified porcine zygote medium (mPZM)] and the additional effects of insulin on in vitro matured and intracytoplasmically inseminated porcine oocytes. In Experiment 1, in vitro matured and intracytoplasmically inseminated porcine oocytes were cultured for 6 days in the following four groups: 1) mPZM-3 (containing bovine serum albumin) + droplet (30 mul), 2) mPZM-3 + WOW, 3) mPZM-4 (containing polyvinyl alcohol) + droplet, and 4) mPZM-4+ WOW. The culture media (mPZM-3 and mPZM-4) and methods (droplet and WOW) did not significantly affect the cleavage rate, but the blastocyst rate of the oocytes cultured in mPZM-3 was significantly (P<0.

Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa.

In this study, two following experiments were performed to improve post-thaw motility and viability of frozen-thawed ram spermatozoa. We examined i) the effects of different concentrations of bovine serum albumin (0, 0.3, 1, 5, 10 and 15% BSA) in semen diluents lacking egg yolk and ii) the effects of four semen diluents, fructose (F: control) and trehalose (T) in semen diluents containing egg yolk, 15% BSA in semen diluents without egg yolk (BSA), and modified phosphate buffered saline (m-PBS).