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81.
82.
Maya OI Keisuke YAMADA Hiroyuki HAYAKAWA Hiroshi SUZUKI 《The Journal of reproduction and development》2013,59(1):92-96
Effective preselection of sex has been accomplished in several species of livestock and
also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting
accuracy is a key prerequisite for the widespread use of sperm sexing. The standard
validation method is flow cytometric remeasurement of the DNA content of the sexed sperm.
Since this method relies on the same instrument that produced the original sperm
separation, it is not truly independent. Therefore, to be able to specifically produce
either male or female offspring in the dog, we developed a method of direct visualization
of sex chromosomes in a single sperm using fluorescence in situ
hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by
immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97%
hybridization efficiency and a good preservation of sperm morphology. There was no
significant difference between the theoretical ratio (50:50) and the observed ratio of X-
and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean
purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for
the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was
evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that
the FISH protocol worked reliably for both unsorted and sexed sperm samples. 相似文献
83.
Mst. Parvin MOSTARI Nahoko IEDA Chikaya DEURA Shiori MINABE Shunji YAMADA Yoshihisa UENOYAMA Kei-ichiro MAEDA Hiroko TSUKAMURA 《The Journal of reproduction and development》2013,59(3):266-272
Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B
(NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on
pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The
present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative
feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine
(nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V)
on LH pulses was determined in ovariectomized (OVX) adult female rats with/without
replacement of negative feedback levels of estradiol (low E2). The mean LH
concentrations and baseline levels of LH secretion in nor-BNI-injected, low
E2-treated rats were significantly higher compared with vehicle-treated
controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse
parameters in OVX rats without low E2 treatment. These results suggest that Dyn
is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The
low E2 treatment had no significant effect on the numbers of ARC
Pdyn (Dyn gene)-,Kiss1- and
Tac2 (NKB gene)-expressing cells. The treatment also did not affect
mRNA levels of Pdyn and Oprk1 (KOR gene) in the
ARC-median eminence region, but significantly increased the ARC kisspeptin
immunoreactivity. These findings suggest that the negative feedback level of estrogen
suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism
without affecting the Dyn and KOR expressions in the ARC. Taken together, the present
result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on
GnRH/LH pulses by reducing the kisspeptin release in female rats. 相似文献
84.
Naoaki Tsutsui Ayano Nagakura-Nakamura Chiaki Nagai Tsuyoshi Ohira Marcy N. Wilder Hiromichi Nagasawa 《Fisheries Science》2013,79(1):33-38
Seven major peptides belonging to the crustacean hyperglycemic hormone family were purified from the sinus gland located in the eyestalk of the kuruma prawn Marsupenaeus japonicus, and their effects on vitellogenin gene expression were examined using the ex vivo ovary incubation system. Six molecular species of crustacean hyperglycemic hormone, Pej-SGP-I, -II, -III, -V, VI, and VII, displayed significant inhibitory effects on vg expression with almost the same efficacies, whereas Pej-SGP-IV (known as molt-inhibiting hormone) did not. Two chromatophorotropic peptides, red pigment-concentrating hormone and pigment-dispersing hormone, which were also present in the sinus glands, did not have a clear effect on the gene expression levels in this incubation system. These results suggest that the six crustacean hyperglycemic hormones are potentially capable of acting as vitellogenesis-inhibiting hormones in M. japonicus. 相似文献
85.
Naoaki Tsutsui Tsuyoshi Ohira Tomoyuki Okutsu Junpei Shinji Sun-Hye Bae Bong Jung Kang Marcy N. Wilder 《Fisheries Science》2013,79(3):357-365
The cDNA encoding a precursor of Liv-SGP-G, the most abundant crustacean hyperglycemic hormone-family peptide in the sinus gland of the whiteleg shrimp Litopenaeus vannamei, was cloned by RT-PCR coupled with 5′- and 3′-RACE. The determined cDNA sequence consisted of 621 nucleotides comprising a 69-bp 5′-untranslated region, a 357-bp open reading frame, and a 195-bp 3′-untranslated region. The deduced sequence of 72 amino acid residues corresponding to mature Liv-SGP-G was completely identical to that of natural Liv-SGP-G, determined in our previous study. The recombinant Liv-SGP-G was thereafter heterologously expressed in Escherichia coli, and its vitellogenesis-inhibiting activity in ex vivo cultured ovarian fragments was examined. The recombinant peptides inhibited vitellogenin gene expression with almost the same efficacy as that of natural Liv-SGP-G purified from sinus glands. 相似文献
86.
87.
Yuuki ISAJI Koki YOSHIDA Hiroshi IMAI Masayasu YAMADA 《The Journal of reproduction and development》2015,61(6):503-510
In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the
nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of
oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum
albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection
medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm
of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in
vitro development of cloned embryos to the blastocyst stage compared with that of a conventional
nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation
rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a
hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments
revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine
12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also
increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of
d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro
and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear
reprogramming. 相似文献
88.
Tagawa M Takeuchi T Fujisawa T Konno Y Yamamoto S Matsumoto K Yokoyama N Inokuma H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(1):99-102
A 2-year-old East Friesian sheep imported from Australia exhibited severe anemia after contagious pustular dermatitis in Hokkaido, Japan. Hemoplasma infection was confirmed in blood smears. Both Mycoplasma ovis and 'Candidatus Mycoplasma haemovis' were detected by PCR and sequence analyses. In the epidemiological analysis, dual pathogens were detected in 6 of 12 (50.0%) sheep imported from Australia with the infected ewe at the same time, 1 of 5 (20.0%) sheep introduced from a domestic farm in Hokkaido, and in 1 of 16 (6.3%) sheep from an epidemiologically unrelated ranch. It is the first clinical case of sheep to confirm coinfection of these pathogens in Japan. 相似文献
89.
Tomohiro Yonezawa Kohei Sato Mona Uchida Naoaki Matsuki Atusi Yamazaki 《Animal Science Journal》2017,88(1):195-200
Contagious yawning is triggered by others yawning, and it has previously been reported in humans, primates and several experimental and companion mammals. Whereas it might be a response to an innate releasing mechanism, contagious yawning is also considered to involve emotional contagion. Here, we demonstrate that sheep, the animal model of livestock animals, also experience contagious yawning. Twelve adult castrated Corriedale sheep were used in this study. Pairs of sheep were adjacently restrained with or without a wooden divider panel to shield them from viewing the other. Their behaviors were video‐recorded for 3 days in each condition. Sheep yawned 2.0 ± 1.1 and 1.2 ± 1.1 times/day/head in the unshielded and shielded conditions, respectively. Unshielded restrained sheep yawned within 1 min after the other one 11.1% of the time, while shielded pairs did not exhibit contagious yawning. Rumination was also highly synchronized under the unshielded condition. These data reveal that contagious yawning and behavioral synchronicity occur in ruminants like sheep, making them a suitable animal model to investigate contagious yawning and the underlying mechanism. 相似文献
90.
Akemi YAMAMOTO Minoru ITOH Yoshikatsu KADOYA Hirokazu KANNO Michi YAMADA Shu FURUYA 《Animal Science Journal》2002,73(4):301-304
An experiment was conducted to determine the effects of supplementing a reduced crude protein (CP) diet with apple pomace on the urinary nitrogen excretion and ammonia emission from slurry in the growing pigs. Four barrows (35 kg bodyweight) were assigned to one of two diets. Each pig was placed in a metabolism cage and fed a low CP, amino acid-supplemented diet (CP 11.33%; low CP diet) or a low CP, amino acid-supplemented diet containing 23.08% dried apple pomace (CP 9.47%; apple pomace diet) for two 14-day experimental periods. After the completion of the first period, the pigs were switched to the other diet. Urine and feces were quantitatively collected daily for 5 days after a 9-day adaptation period. The daily nitrogen intake for the low CP diet and the apple pomace diet was 17.76 g/pig and 18.64 g/pig, respectively. The pigs fed the apple pomace diet excreted more fecal nitrogen (6.86 g/day) than the pigs fed the low CP diet (3.63 g/day) ( P < 0.001), but urinary nitrogen excretion with the apple pomace diet was 3.11 g/day, which was much lower than that for the low CP diet (5.95 g/day) ( P < 0.001). The daily ammonia emission from the mixture of urine and feces determined by an in vitro method was much lower for pigs fed the apple pomace diet (120 mg) than that for pigs fed the low CP diet (603 mg) ( P < 0.01). The addition of apple pomace to a reduced CP, amino acid-supplemented diet reduces urinary nitrogen excretion and thereby ammonia emission. 相似文献