Introduction of a point mutation in the KRAS gene of in vitro fertilized porcine zygotes via electroporation of the CRISPR/Cas9 system with single-stranded oligodeoxynucleotides     

Manita Wittayarat  Maki Hirata  Zhao Namula  Yoko Sato  Nhien T. Nguyen  Quynh A. Le  Qingyi Lin  Koki Takebayashi  Fuminori Tanihara  Takeshige Otoi 《Animal Science Journal》2021,92(1):e13534

This study aimed to investigate the efficiency of KRAS gene editing via CRISPR/Cas9 delivery by electroporation and analyzed the effects of the non-homologous end-joining pathway inhibitor Scr7 and single-stranded oligodeoxynucleotide (ssODN) homology arm length on introducing a point mutation in KRAS. Various concentrations (0–2 µM) of Scr7 were evaluated; all concentrations of Scr7 including 0 µM resulted in the generation of blastocysts with a point mutation and the wild-type sequence or indels. No significant differences in the blastocyst formation rates of electroporated zygotes were observed among ssODN homology arm lengths, irrespective of the gRNA (gRNA1 and gRNA2). The proportion of blastocysts carrying a point mutation with or without the wild-type sequence and indels was significantly higher in the ssODN20 group (i.e., the group with a ssODN homology arm of 20 bp) than in the ssODN60 group (gRNA1: 25.7% vs. 5.4% and gRNA2: 45.5% vs. 5.9%, p < .05). In conclusion, the CRISPR/Cas9 delivery with ssODN via electroporation is feasible for the generation of point mutations in porcine embryos. Further studies are required to improve the efficiency and accuracy of the homology-directed repair.  相似文献   

Effect of K-type and Ca-type artificial zeolites applied to high sodic soil on the growth of plants different in salt tolerance     

Mina YAMADA  Hideyasu FUJIYAMA  Tsuneyoshi ENDO  Maki Uehira RIKIMARU  Yoko SASAKI  Sadahiro YAMAMOTO  Toshimasa HONNA  Tahei YAMAMOTO 《Soil Science and Plant Nutrition》2007,53(4):471-479

  相似文献   

Abnormal functions of Leydig cells in crossbred cattle–yak showing infertility     

Yoko Sato  Ryota Kuriwaki  Shiki Hagino  Megumi Shimazaki  Rentsenkhand Sambuu  Maki Hirata  Fuminori Tanihara  Mitsuhiro Takagi  Masayasu Taniguchi  Takeshige Otoi 《Reproduction in domestic animals》2020,55(2):209-216

In Mongolia, yak (Bos grunniens) are able to live in alpine areas and their products greatly influence the lives of the local people. Increased vigour in hybridized yak and cattle can offer benefits for livestock farmers. However, male hybrids show reproductive defects resulting from spermatogenesis arrest, affecting the conservation and maintenance of dominant traits in the next generation. The underlying mechanisms involved in hybrid cattle–yak infertility have recently been investigated; however, the genetic cause is still unclear. Androgens and androgen receptor (AR) signalling are required for spermatogenesis. We, therefore, evaluated the expression of AR, 3β-hydroxysteroid dehydrogenase (3βHSD) and 5α-reductase 2 (SRD5A2) in Leydig cells to investigate their function in cattle–yak spermatogenesis. Testicular tissues from yaks (1–3 years old) and hybrids (F1–F3, 2 years old) were collected and subjected to immunohistochemistry and image analyses to investigate the expression of each parameter in the Leydig cells. After maturation at 2 years, the expression levels of AR increased and the levels of 3βHSD decreased, but the SRD5A2 levels remained constant in yak. However, the cattle–yak hybrid F2 showed immature testicular development and significantly different expression levels of AR and 3βHSD compared with mature yak. These results suggest that the decreased expression of AR and increased expression of 3βHSD in the Leydig cells of cattle–yak hybrid testes may represent one of the causes of infertility. Our study might help in solving the problem of infertility in crossbreeding.  相似文献   

Assessment of repeatability of a wireless, inertial sensor-based lameness evaluation system for horses   总被引：1，自引：0，他引：1  

Keegan KG  Kramer J  Yonezawa Y  Maki H  Pai PF  Dent EV  Kellerman TE  Wilson DA  Reed SK 《American journal of veterinary research》2011,72(9):1156-1163

  相似文献   

A case of Iriomote cat (Prionailurus bengalensis iriomotensis) with Hepatozoon felis parasitemia     

Sakuma M  Nishio T  Nakanishi N  Izawa M  Asari Y  Okamura M  Shimokawa Miyama T  Setoguchi A  Endo Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(10):1381-1384

We herein present clinical findings of an Iriomote cat with Hepatozoon felis parasitemia. A male Iriomote cat was captured for ecological analyses three times from January 2010 to January 2011. Although this cat did not show any hematological abnormalities at the time of the first capture, H. felis parasitemia and increased serum creatine kinase levels were detected at the second and third captures. H. felis infection was confirmed by polymerase chain reaction, and amplified 18S ribosomal RNA gene fragments were 100% identical to those of H. felis in leopard cats in Korea. Although the virulence of H. felis in this cat was suggested to be low, this is the first report of an H. felis-infected Iriomote cat with parasitemia.  相似文献   

A "possible" involvement of TNF-alpha in apoptosis induction in peripheral blood lymphocytes of cats with feline infectious peritonitis     

Takano T  Hohdatsu T  Hashida Y  Kaneko Y  Tanabe M  Koyama H 《Veterinary microbiology》2007,119(2-4):121-131

Feline infectious peritonitis (FIP) cats show a decrease in peripheral blood lymphocyte counts, and a particularly marked decrease in T cells including CD4+ and CD8+ cells. In this study, we showed that lymphopenia observed in FIP cats was due to apoptosis, and that the ascitic fluid, plasma, and culture supernatant of peritoneal exudate cells (adherent cells with macrophage morphology, or PEC) from FIP cats readily induced apoptosis in specific pathogen-free cat peripheral blood mononuclear cells, particularly CD8+ cells. In addition, TNF-alpha released from macrophages and TNF-receptor (TNFR) 1 and TNFR2 mRNA expression in lymphocytes were closely involved in this apoptosis induction. In particular, in CD8+ cells cultured in the presence of the PEC culture supernatant, the expression levels of TNFR1 and TNFR2 mRNA were increased, indicating that CD8+ cells are more susceptible to apoptosis induction by TNF-alpha than other lymphocyte subsets, particularly B cells (CD21+ cells). The results of this study suggest that TNF-alpha, produced by virus-infected macrophages, is responsible for induction of apoptosis in uninfected T cells, primarily CD8+ T cells.  相似文献   

Pancreatic torsion in a dog     

Tamera L. Brabson  Lynn C. Maki  Susan M. Newell  S. Christopher Ralphs 《The Canadian veterinary journal. La revue veterinaire canadienne》2015,56(5):476-478

A 6-month-old male intact Cane Corso mastiff dog was presented for a recent history of vomiting, abdominal pain, and lethargy. A diagnosis of pancreatic torsion was made during abdominal exploratory surgery and was confirmed with histopathology. The dog underwent partial pancreatectomy and recovered with no complications.  相似文献   

Melatonin protects the integrity of granulosa cells by reducing oxidative stress in nuclei,mitochondria, and plasma membranes in mice     

Manabu TANABE  Hiroshi TAMURA  Toshiaki TAKETANI  Maki OKADA  Lifa LEE  Isao TAMURA  Ryo MAEKAWA  Hiromi ASADA  Yoshiaki YAMAGATA  Norihiro SUGINO 《The Journal of reproduction and development》2015,61(1):35-41

Melatonin protects luteinized granulosa cells (GCs) from oxidative stress in the follicle during ovulation. However, it is unclear in which cellular components (e.g., nuclei, mitochondria, or plasma membranes) melatonin works as an antioxidant. GCs from immature (3 wks) ICR mice were incubated with hydrogen peroxide (H2O2; 0.01, 0.1, 1, 10 mM) in the presence or absence of melatonin (100 μg/ml) for 2 h. DNA damage was assessed by fluorescence-based immunocytochemistry using specific antibodies for 8-hydroxydeoxyguanosine (8-OHdG), an indicator of oxidative guanine base damage in DNA, and for histone H2AX phosphorylation (γH2AX), a marker of double-strand breaks of DNA. Mitochondrial function was assessed by the fluorescence intensity of MitoTracker Red probes, which diffuse across the membrane and accumulate in mitochondria with active membrane potentials. Lipid peroxidation of plasma membranes was analyzed by measuring hexanoyl-lysine (HEL), a oxidative stress marker for lipid peroxidation. Apoptosis of GCs was assessed by nuclear fragmentation using DAPI staining, and apoptotic activities were evaluated by caspase-3/7 activities. H2O2 treatment significantly increased the fluorescence intensities of 8-OHdG and γH2AX, reduced the intensity of MitoTracker Red in the mitochondria, increased HEL concentrations in GCs, and enhanced the number of apoptotic cells and caspase-3/7 activities. All these changes were significantly decreased by melatonin treatment. Melatonin reduced oxidative stress-induced DNA damage, mitochondrial dysfunction, lipid peroxidation, and apoptosis in GCs, suggesting that melatonin protects GCs by reducing oxidative stress of cellular components including nuclei, mitochondria, and plasma membranes. Melatonin helps to maintain the integrity of GCs as an antioxidant in the preovulatory follicle.  相似文献   

The influences of hyperbaric oxygen therapy with a lower pressure and oxygen concentration than previous methods on physiological mechanisms in dogs     

Maki ISHIBASHI  Akiyoshi HAYASHI  Hideo AKIYOSHI  Fumihito OHASHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(3):297-304

Recently, hyperbaric oxygen therapy with a lower pressure and oxygen concentration (L-HBOT) than previous methods has been used for dogs in Japan; however, the influences of L-HBOT on dogs have not been clarified. To verify the influences of L-HBOT on physiological mechanism in dogs, we investigated blood gas parameters, glutathione peroxidase (GPx) activity, heart rate variability, stress-related hormones and skin conductance (SC) in 4 clinically normal beagle dogs with catheters in their carotid arteries and jugular veins when they were quiet, after running, after receiving L-HBOT (30% oxygen concentration, 1.3 atmospheres absolute, 30 min) or after not receiving L-HBOT. The results showed there were no changes in blood gas parameters, heart rate variability and catecholamine levels after L-HBOT. GPx activity was significantly higher, and the SC and cortisol level were lower in dogs that received L-HBOT than those when they were quiet. These results suggested that L-HBOT may have a small influence on oxygenation dynamics, activate antioxidant enzymes such as GPx, restrain autonomic nervous activity and control the balance between oxidation and antioxidation inside the body.  相似文献   

Sex and age differences in meat composition of Yeso sika deer (Cervus nippon yesoensis) reared for a short period after capture in the wild          下载免费PDF全文

Maki Hayashida  Kousaku Souma  Kazuki Sugo  Shin‐ichi Araki  Fumiaki Ishizaka  Masami Ueda  Takamasa Kasai  Takayoshi Masuko 《Animal Science Journal》2015,86(2):207-213

Yeso sika deer captured in winter around Lake Akan in Hokkaido were reared for 8–10 months at Tokyo University of Agriculture in Abashiri. Six 1‐year‐old females and males and six 2‐year‐old or older (adult) females and males were slaughtered and their carcasses were processed. The chemical composition, mineral contents and fatty acid composition of the loin were measured. No marked influence of gender or age was noted in the chemical composition of loin. In the mineral contents, significant differences were noted. The potassium and sulfur contents were lower and the sodium content was higher in adult deer meat (P < 0.05, respectively) and the potassium content was higher in male deer meat (P < 0.05). Arsenic, cadmium or lead were not detected. In the unsaturated fatty acid, a significant interaction was detected (P < 0.05), and it was high in 1‐year‐old female deer meat and low in 1‐year‐old male deer meat. Significant gender or age differences were noted only in the mineral contents in the loin of deer reared for a short period after capture.  相似文献