Expression and localisation of claudin-1, -2, -3, -4, -5, -7 and -10 proteins in the normal canine mammary gland

The recently identified claudins are dominant components of tight junctions, responsible for cell adhesion, polarity and paracellular permeability. Certain claudins have been shown to have relevance in tumour development. The aim of the present study was to analyse the expression of claudin-1, -2, -3, -4, -5, -7 and -10 in normal canine mammary glands. Samples from the inguinal mammary regions of 20 non-castrated, 1-13 years old female dogs were studied. Immunohistochemical analysis was performed on conventional specimens and tissue microarrays. The results of the immunohistochemical reactions detecting claudins in tissue sections were photodocumented. The immunoreactivity of claudins was quantitatively analysed on digital images using Leica QWin morphometry software. Intense membranous immunolabelling was found for claudin-1, -3 and -7, intense membranous with non-granular cytoplasmic immunolabelling for claudin-2, moderate membranous immunolabelling for claudin-4 and -5, and weak membranous immunolabelling for claudin-10. The occurrence of tight junctions was confirmed by ultrathin section electron microscopy. The available data suggested that claudins might be proteins preserved throughout the evolution of mammals. The results of our study support the concept that they are indeed preserved, since the same type of claudins, in identical distribution, could be detected in our canine mammary tissue samples as could be found in human mammary tissue.     

Comparative Immunolocalization of Heat Shock Proteins (Hsp)-60, -70, -90 in Boar, Stallion, Dog and Cat Spermatozoa

Heat shock proteins (Hsp)-60, -70 and -90 are important testis chaperones that fulfil several functions during sperm cell maturation. In post-meiotic cells, their expression may change or may be undetectable and in some species it may be evident in mature spermatozoa. The aims of this study were to verify whether Hsp60, -70 and -90 are present in the sperm, and to compare their localization in boar, stallion, cat and dog spermatozoa by immunofluorescence. Hsp-60 immunoreactivity was detected in sperm midpiece in all the species examined. In stallion sperm, Hsp70 signal was localized in the sub-equatorial band, whereas immunoreactivity was evident on the neck of dog spermatozoa and on both neck and sub-equatorial region of cat spermatozoa. In agreement with our previous observations, a triangular fluorescent signal in the equatorial segment of fresh boar sperm was detected. Hsp90 immunoreactivity was present in different portions of sperm tail: in the midpiece of both boar and cat spermatozoa and in the neck and throughout the tail in dog and stallion spermatozoa, respectively. When capacitation and acrosome reaction were induced in boar, stallion and dog spermatozoa, no changes in both Hsp60 and -90 were recorded by either Western blot or immunofluorescence. After induction of acrosome reaction, a Hsp70 redistribution in boar spermatozoa and an increased percentage of stallion spermatozoa showing the post-acrosomal signal were observed although no changes were recorded by Western blot; in dog spermatozoa, no changes in Hsp70 were found by Western blot and immunofluorescence after capacitation and acrosome reaction.     

德国汉诺威国际畜牧展报告

<正>上周我们参加了德国汉诺威国际畜牧展。我们看到:德国汉诺威国际畜牧展两年举办一次,展会所在的大展览中心有超过20个大型建筑。这是我们参加过的最大的畜禽展。有数百家参展商。有很多非常大的展位,特别是一些养猪设备公司。我们估计有些公司花了上百万美元来参展并布置展位。有一些令人印象深刻的展位,似乎养猪设备行业展开了军备竞赛来布置更大更漂亮的展位方面。必达公司为其它公司设置了高门槛。     

草原畜牧展

<正>上周,我们参加了在马尼托巴省温尼伯举办的草原畜牧展。这是首届草原畜牧展,这个展会的前身为马尼托巴养猪日。草原畜牧展是多畜种的活动,不仅有猪业人士参展,还有牛业和禽业等。活动在温尼伯附近的维多利亚酒店会议中心举行。以下是我们的观察:维多利亚酒店的会议场所比温尼伯市区会议中心或布兰登中心更好。演讲者的区域位于会议区,演讲者的声音带来了噪音。我们希望演讲者能发现四处走动的人们所带来的影响。在我们参加的活动中,这是首次会议区没有食物的。     

犬瘟热病毒的蚀斑克隆试验

对血清、胰酶以及染色时间等可能影响犬瘟热病毒蚀斑形成的条件进行了摸索。结果发现,对必须同步接毒的CDV小熊猫（LP）株来讲,其形成蚀斑的最佳条件是：同步接毒10h后,加入血清含量的2%的无酚红的DMEM营养琼脂,第5d时,加入1/5体积的胰酶含量为1/8000的含中性红的DMEM营养琼脂染色,于第6d时,便可形成蚀斑。挑选不同大小的蚀斑,克隆3次后,获得了蚀斑直径为0.4～0.6,0.6～0.8及0.7～0.9mm的克隆株各一个。     

microRNA-1，-133，-206在鼠肌肉中的表达谱

MicroRNAs（miRNAs）是一类重要的调节基因转录的非编码小RNA。其中miR-1、miR-133、miR-206是肌肉特异表达的microRNA,同属miR-1家族。它们在胚胎发育时期的表达及作用已有很多文章阐述,但很少有文章详细分析它们在成体中的表达。为检测这3类microRNA的表达差异,选择BALB/c品系小鼠的股二头肌,提取RNA。设计microRNA专用的茎环状引物,通过实时定量PCR,检验miR-1、miR-133、miR-206和miR-181a在骨骼肌组织中的表达情况。结果发现,miR-133的表达量要明显高于miR-1和miR-206,miR-1略高于miR-206的表达量,作为对照,miR-181a的表达量非常微小。分析该现象,根据资料和网络软件预测,绘制这3类microRNA对肌肉生长发育作用的关系图,初步解释各种microRNA之间呈现表达差异的原因。为下一步研究成体中microRNA及其靶位点提供依据和方向。