Detection of APAF1 mutation in Holstein cows and mummified foetuses in Japanese dairy herds

Some of the highest genetic merit sires have been shown to harbour recessive mutations affecting fertility, which may spread rapidly in the population through AI. These disorders may result in abortion and decline in pregnancy per insemination in cows. This study was carried out on 240 Holstein‐Friesian cows and 15 mummified foetuses. Blood and tissue samples were collected from the cows and mummified foetuses, respectively, for DNA extraction. Allele‐specific PCR was designed for the detection of the cows and foetuses carrying the nonsense mutation (C/T) in apoptosis peptide activating factor 1 gene (APAF1). The mutant allele frequency of the APAF1 in carrier cows and mummified foetuses was calculated. Milk samples were taken from the carrier and non‐carrier cows for progesterone assay. The allele‐specific PCR reaction efficiently distinguished the C/T mutation in APAF1. Of 240 cows, seven cows (2.9%) were diagnosed to carry one copy of the mutant allele of APAF1. However, the carrier frequency was 33.3% in mummified foetuses (five of 15). The mutant allele frequency was 0.02 and 0.17 in the cows and mummified foetuses, respectively. Concentrations of progesterone did not differ between cows with APAF1 mutation and non‐carrier cows during 45 days post‐insemination. This study provided allele‐specific PCR for the detection of APAF1 mutation in cows. Moreover, it reports the carrier and mutant allele frequencies of APAF1 in dairy cows and mummified foetuses in Japan.     

Detection of the Matrix Metalloproteinases MMP‐2 and MMP‐9 and Tissue Inhibitors of Metalloproteinases TIMP‐1 and TIMP‐2 in Llama (Lama glama) Oviduct

Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are involved in several reproductive events like oocyte–spermatozoa interaction and semen liquefaction. In order to study their role in the llama oviductal reproductive process, MMP activity in oviductal fluid (OF) was assayed. Considering that llama genome sequences are partially known, a strategy to procure cDNA sequences of MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 was designed. Afterwards, their expression patterns in the different llama oviductal segments were assayed. Gelatine zymograms detected 62 and 94 kDa protease activities that matched MMP‐2 and pro‐MMP‐9, respectively. Expression pattern analysis showed that MMP and TIMP mRNAs were present in ampulla, isthmus, utero‐tubal junction (UTJ) and papilla. Altogether, these findings support the argument that MMPs/TIMPs are produced in the oviduct and secreted into the oviductal lumen. Our results encourage further studies to elucidate the role of these proteins in reproductive oviductal events.     

乌冈栎种群天然更新研究

种群更新对维持种群的稳定和发展具有极其重要的意义。通过野外实地调查,对不同地区乌冈栎种群的更新情况进行了分析,结果如下:1)大多数地区乌冈栎种群的幼苗结构呈倒金字塔型,说明乌冈栎种群的更新存在一定的困难;但在乌冈栎顶级群落中,由于仍存在一定数量的幼树,短期内的更新仍能继续进行;2)从乌冈栎幼苗空间分布格局来看,林窗或林缘处呈聚集分布,而林冠下呈均匀分布,这与乌冈栎幼苗生长的需光性有极大的关系;3)乌冈栎的萌发实验表明,1～2个月的沙藏处理有利于提高萌发率,而萌发过程中鼠害和虫蛀是限制出苗率的主要原因之一;4)乌冈栎种子的传播除靠重力外,啮齿类动物对它的影响最大,他们之间存在一定的权衡关系。为了提高乌冈栎种群的天然更新率,适当干扰地开辟林窗是较佳的途径。