Tyrosine phosphorylation of a 38-kDa capacitation-associated buffalo (Bubalus bubalis) sperm protein is induced by L-arginine and regulated through a cAMP/PKA-independent pathway

The aim of the present study was to determine the effect of L-arginine on nitric oxide (NO*) synthesis, capacitation and protein tyrosine phosphorylation in buffalo spermatozoa. Ejaculated buffalo spermatozoa were capacitated in the absence or presence of heparin, or L-arginine or N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS) for 6 h. Capacitating spermatozoa generated NO* both spontaneously and following stimulation with L-arginine and L-NAME quenched such L-arginine-induced NO* production. Immunolocalization of NOS suggested for existence of constitutive NOS in buffalo spermatozoa. L-Arginine (10 mm) was found to be a potent capacitating agent and addition of L-NAME to the incubation media attenuated both L-arginine and heparin-induced capacitation and suggested that NO* is involved in the capacitation of buffalo spermatozoa. Two sperm proteins of M(r) 38 000 (p38) and 20 000 (p20) were tyrosine phosphorylated extensively by both heparin and L-arginine. Of these, the tyrosine phosphorylation of p38 was insensitive to both induction by cAMP agonists as well as inhibition by a protein kinase A (PKA) inhibitor. Further, most of these L-arginine-induced tyrosine phosphorylated proteins were localized to the midpiece and principal piece regions of flagellum of capacitated spermatozoa and suggested that sperm flagellum takes active part during capacitation. These results indicated that L-arginine induces capacitation of buffalo spermatozoa through NO* synthesis and tyrosine phosphorylation of specific sperm proteins involving a pathway independent of cAMP/PKA.     

H2O2浸渍处理对水牛角胎力学特性的影响

目的将水牛角胎制成公称尺寸试件。研究脱蛋白处理水牛角胎后,其抗原性和生物力学强度与 30％H2O2处理的时间关系;选择最大限度降低角胎的抗原性和又尽量保留其生物力学强度的最佳时间。方法用氯仿-甲醇1:1浸渍48小时,再用30％H2O2浸渍,时间分别为0小时,24小时,48小时,用电测试法进行弹性模量和用直接加压法测其抗压强度。结果试验发现30％H2O2浸渍0小时组、24小时组和48小时组弹性模量没有显著差异P>0．05;抗压强度24小时组和48小时组明显低于0小时组;而24 小时组和48小时组没有显著性差异。结论试验提示: 水牛角胎经30％H2O2浸渍48小时处理,已能使其抗原性降至较低水平,并较好地保留了其生物力学强度。     

FISH mapping of theα-S2 casein gene on river buffalo and cattle chromosomes identifies a nomenclature discrepancy in the bovine karyotype

A mixture of five genomic clones spanning the-S2 casein gene (CSN1S2) were mapped to river buffalo (Bubalus bubalis L.) and cattle (Bos taurus L.) chromosomes by fluorescencein situ hybridization (FISH) and R-banding. Clear probe hybridization signals were detected on river buffalo chromosome 7q, band 32, and the homologous cattle chromosome. These mapping data allow the indirect assignment of the entire cattle U15 syntenic group to river buffalo chromosome 7. The assignment of U15 to river buffalo chromosome 7 is discussed in the light of chromosomal nomenclature discrepancies involving the homologous cattle chromosome.accepted for publication by D. Ward     

Comparative FISH-mapping of villin (VIL) gene in river buffalo, sheep and goat chromosomes

Two genomic clones of the villin (VIL) gene were independently hybridized on river buffalo (Bubalus bubalis, BBU), sheep (Ovis aries, OAR) and goat (Capra hircus, CHI) chromosomes by using sequential fluorescence in situ hybridization (FISH) and R-banding (RBP- and RBA-banding). Clear hybridization signals revealed that VIL is located in BBU 2q33, OAR 2q33 and CHI 2q33. These chromosomes and chromosome bands are believed to be homologous and the VIL locus is the same as that previously found on cattle chromosome 2q43. VIL localization in these three species allows us tentatively to assign all cattle U17 to BBU and CHI 2q and to extend the physical map to OAR 2q.This revised version was published online in November 2005 with corrections to the Cover Date.     

Effect of calmodulin-like protein from buffalo (Bubalus bubalis) seminal plasma on Ca2+, Mg2+-ATPase of purified plasma membrane of buffalo spermatozoa

Summary. Buffalo sperm heads and tails were cleaved by sonication and isolated in relatively pure proportions i.e. 95% and 98% respectively, by discontinuous sucrose density-gradient centri-fugation. Purified plasma membranes from the isolated sperm heads and tails were obtained by hypotonic treatment and brief sonication followed by discontinuous sucrose density-gradient centrifugation. Ca²⁺, Mg²⁺-ATPase activity was evident in plasma membrane from sperm heads and tails, although activity was greater in the latter. A calmodulin-like protein isolated from buffalo seminal plasma increased the Ca²⁺, Mg²⁺-ATPase of plasma membrane from the sperm heads and tails by 128 and 136% respectively. Based upon the data obtained here and elsewhere (Sidhu & Guraya, 1989a) a model is proposed which explains regulation of Ca²⁺ in buffalo spermatozoa and implicates calmodulin-like protein and Ca²⁺, Mg²⁺-ATPase in sperm acrosome reaction.     

水中草颗粒对实验动物复发性口腔溃疡红细胞免疫功能的研究

目的:探讨水中草颗粒对复发性口腔溃疡红细胞免疫功能的影响。方法:采用免疫方法建立复发性口腔溃疡动物模型,将成模的复发性口腔溃疡家兔随机分为模型对照组、左旋咪唑组、水中草低剂量组和高剂量组,同时设立正常对照组。从实验的第7周开始,左旋咪唑组、水中草低剂量组及水中草高剂量组每日分别以相应药物灌胃,共连续28天。最后根据红细胞C3b受体花环试验和免疫复合物花环试验检测红细胞免疫功能。结果:模型对照组家兔红细胞C3b受体花环率显著降低,免疫复合物花环率显著增高,给予左旋咪唑、水中草颗粒后,红细胞C3b受体花环率显著升高,红细胞免疫复合物花环率显著降低。结论:水中草颗粒能够提高红细胞C3b受体花环率及降低红细胞免疫复合物花环率。     

水牛成纤维细胞核移植的研究

本研究对影响水牛耳部成纤维细胞核移植效果的因素进行了探讨。体外成熟 2 2h的水牛卵母细胞经显微操作去核后 ,将一个经传代培养的水牛耳部成纤维细胞注入到胞质内。经核移植的水牛卵母细胞用 5μM离子霉素激活处理 5min ,然后在含有 2mM 6 -甲二氨基嘌呤 ( 6 -DMAP)的培养液培养 3h。当成纤维细胞用DNA合成抑制剂Aphidicolin( 0 .4μg/ml)培养 1d或 2d再进行核移植时 ,其囊胚发育率 ( 2 .0 %和 0 )明显低于 ( p <0 .0 5)对照组 ( 4.9% ) ,虽然其分裂率 ( 6 0 .9%和 6 0 .5% )与对照组 ( 6 2 .9% )无明显差异 ( p >0 .0 5)。当成纤维细胞的培养代数由G5增加到G6和G7时 ,囊胚发育率亦随之下降 ( 5.3% ,2 .0 %和 1 .8% ,p <0 .0 5) ,但分裂率无显著差异 ( 6 8.0 % ,6 0 .9%和 6 0 .7% ,p >0 .0 5)。结果表明 ,在现有核移植实验条件下 ,水牛成纤维细胞的培养代数应控制在 5代之内 ,且不必用DNA合成抑制剂对其进行周期调控     

重组日本血吸虫26 kDa GST抗原免疫水牛后抗体动态及免疫保护性效果

目的　观察重组日本血吸虫 2 6kDaGST抗原 (reSjc2 6GST)免疫役用放养水牛 (简称水牛 )后抗体动态及免疫保护性的效果。　方法　试验组 96头水牛 ,用reSjc2 6GST免疫 3次 ,每次间隔 2wk ,3次剂量分别为 0 2、 0 2和 0 1mg。对照组 90头水牛不作免疫 ;观察 2组水牛免疫前及免疫后 2、 5、 9、 12、 15和 2 0个月的抗体水平及血吸虫感染率的变化。　结果　试验组机体产生特异性抗reSjc2 6GST抗体 ,其抗体水平呈明显的梯形升高趋势。试验组免疫后 2 0个月血吸虫感染率比免疫前下降了 62 2 % ,比同期对照组低 67 7%。　结论　用reSjc2 6GST免疫水牛能产生特异性抗体 ,在免疫后 2 0个月内维持较高水平 ,有一定的抗血吸虫自然感染的保护力。     

African Buffalo Movement and Zoonotic Disease Risk across Transfrontier Conservation Areas,Southern Africa

We report on the long-distance movements of subadult female buffalo within a Transfrontier Conservation Area in Africa. Our observations confirm that bovine tuberculosis and other diseases can spread between buffalo populations across national parks, community land, and countries, thus posing a risk to animal and human health in surrounding wildlife areas.