Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.     

Gene dropping analysis of founder contributions in a closed Japanese black cattle population

Gene dropping simulation was applied to Japanese Black cattle population in Hyogo prefecture, to examine the survivals of alleles originated from founder animals. In the analysis, unique alleles were assigned to founders, and the genotypes of all descendants along the actual pedigree were generated through Monte Carlo simulation following Mendelian segregation rules. By replicating this process 10 000 times, the distribution of frequencies of alleles from each founder was estimated. From the distribution, several quantities useful for the management of genetic diversity, such as the probability of allele extinction and the probability of alleles surviving at a critically low frequency were derived. The materials used were 68 781 animals born in 1955–1998 and their pedigree records traced back to the population in 1937 or before. The expected number of alleles retained in the population drastically decreased during the analyzed period, and reached to 57.9 in the population of 1998, which was only 3.3% of the total number of alleles assigned to founders. Detailed analysis of major founders with relatively high genetic contributions to the current population revealed that alleles from most of the major founders are now at high risk of future extinction. These results strongly suggest that for the management of genetic diversity, the genetic contributions of founders are not fully informative, and emphasize the importance of the detection of live animals having founder alleles with high extinction possibilities.     

粳稻食味品质改良研究现状与展望

进入21世纪,我国北方特别是东北粳稻面积不断扩大,单产明显提高,品质稳定改善,改良食味品质是今后重要研究内容。本文主要从食味品质鉴定方法,稻米淀粉种类和含量及分子结构、蛋白质种类和含量、其他物质含量与食味品质的关系,饭粒表面和内部超微结构与食味品质的关系,食味品质相关性状遗传分析与育种应用等方面介绍了日本粳稻食味品质研究现状,认为结合东北粳稻实际深入开展食味品质育种、栽培和生物技术等应用基础研究,充分利用籼稻二次枝梗粒偏向穗轴上部分布、穗颈大维管束数和一次枝梗数多等有利基因,同时减少对食味品质的不良影响,是今后北方尤其是东北籼粳稻杂交优质高产育种的重要研究课题。     

鲢肌球蛋白重链球状结构域的cDNA克隆及结构解析

根据已经获得的两种鲢肌球蛋白重链同工型基因(低温型sc-w和高温型sc-s)在3′端展现的明显差异,设计了2个特异性的反向引物,以鲤科鱼类肌球蛋白重链5′端的保守序列为正向引物,通过Long-PCR对编码鲢两种肌球蛋白重链同工型的球状结构域(Subfragment-1,S1)的全长基因进行了克隆和测序,并推断出它们一级结构的氨基酸序列。研究结果表明,sc-w与sc-s在S1的初级结构上显示80.5%的同源性、与已经报道的草鱼低温型(gc10)有97.2%的高同源性;sc-s则与草鱼中间型(gcI)和高温型(gc30)显示了分别为98.4%和97.1%的高同源性。低温型的sc-w和gc10在S1初级结构上展现的特有变异主要发生在43个氨基酸残基位点,其中15个属保守性残基。对S1区域中两个功能性的表面环loop1(与ATP结合位点有关)和loop2(与肌动蛋白结合位点有关)的结构解析发现,sc-w和gc10在两个表面环的长度、残基电荷分布和氨基酸组成等方面与其它同工型之间存在明显差异,揭示了这两个表面环的结构差异可能影响了栖息于不同环境温度下的淡水鱼的肌球蛋白分子马达功能。分子系统树的分析结果进一步证明,鱼类栖...     

Piebald mutation on a C57BL/6J background

The classic piebald mutation in the endothelin receptor type B (Ednrb) gene was found on rolling Nagoya genetic background (PROD-s/s) mice with white coat spotting. To examine whether genetic background influenced the phenotype in the piebald mutant mice, we generated a congenic strain (B6.PROD-s/s), produced by repeated backcrosses to the C57BL/6J (B6) strain. Although B6.PROD-s/s mice showed white coat spotting, 7% of B6.PROD-s/s mice died between 2 and 5 weeks after birth due to megacolon. The PROD-s/s, s/s and Japanese fancy mouse 1 (JF1) strains, which also have piebald mutations on different genetic backgrounds with B6, showed only pigmentation defects without megacolon. In expression analyses, rectums of B6.PROD-s/s with megacolon mice showed ~5% of the level of Ednrb gene expression versus B6 mice. In histological analyses, aganglionosis was detected in the rectum of megacolon animals. The aganglionic rectum was thought to lead to severe constipation and intestinal blockage, resulting in megacolon. We also observed an abnormal intestinal flora, including a marked increase in Bacteroidaceae and Erysipelotrichaceae and a marked decrease in Lactobacillus and Clostridiales, likely inducing endotoxin production and a failure of the mucosal barrier system, leading ultimately to death. These results indicate that the genetic background plays a key role in the development of enteric ganglion neurons, controlled by the Ednrb gene, and that B6 has modifier gene (s) regarding aganglionosis.     

Histopathological changes in the pancreas due to decreased pancreatic blood flow in a canine tachycardia-induced cardiomyopathy model

In dogs, pancreatic acinar cell injury is thought to be caused by decreased pancreatic blood flow due to heart failure. In previous our report, it demonstrated that decreased heart function causes a significant decrease in pancreatic blood flow in heart failure dog model caused by rapid ventricular pacing (RVP). However, the types of histopathological changes remain unclear. We aimed to verify the types of histopathological changes occurring in the pancreatic tissue due to decreased heart function. After RVP for 4 weeks, atrophy of pancreatic acinar cells, characterized by a decrease in zymogen granules, was observed in all areas of the pancreas. In conclusion, the result of this study suggests that attention should be paid to ischemia/hypoperfusion injury in the pancreas.