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1.
试验采用两步酶消化法对2月龄公羔睾丸进行处理,得到了总数为7.44×109的生精细胞悬液,经台盼蓝染色,细胞活率在90%以上;支持细胞的贴壁性极好,生精细胞附着在支持细胞上,随着培养时间延长,生精细胞、支持细胞逐渐退化,呈现“集合”的趋势,细胞数量逐渐减少;经BrdU标记、H.E.染色,检测培养的生精细胞到分化到圆形精子细胞阶段,表明成功构建了山羊睾丸生精细胞共培养体系,可为后续深入研究山羊精子体外成熟培养提供一定的实验依据.  相似文献   

2.
采用玻璃化冷冻方法对野外采集的牦牛睾丸组织进行冷冻,通过HE(hematoxylin-eosin)染色分析发现玻璃化冷冻后的牦牛睾丸组织曲细精管结构保存较完好,曲细精管可见大量形态完好的各类生精细胞。采用台盼蓝染色检测细胞活率,发现冷冻复苏后细胞活率可达80.20%。分别通过生精细胞和精原干细胞标志蛋白DDX4和GFRA1免疫荧光染色发现复苏后培养14天后的生精细胞和精原干细胞的数量明显减少。通过RT-qPCR对复苏后不同实验处理组牦牛睾丸细胞标志基因的表达分析,发现培养30天的生精细胞中的精原干细胞标志基因Thy1和UCHL1的表达量显著增高。因此,玻璃化冷冻保存的牦牛睾丸组织中曲细精管及其生精细胞得到了较好的保护,该方法对于其他哺乳动物生精细胞的长久有效保存具有重要的参考价值。  相似文献   

3.
探讨了在二乙基己烯雌酚(DES)诱发成年动物生精细胞凋亡过程中睾丸一氧化氮(NO)生成和生精细胞一氧化氮合酶(eNOS和iNOs)表达的变化,以期为阐明DES诱发生精细胞凋亡机理的研究提供基础资料。成年雄性仓鼠皮下注射不同剂量DES(分别为0.01、0.1和1mg/kg体重),连续注射7d后取其睾丸,进行NO含量的测定和eNOS、iNOS免疫组化染色。电镜观察生精细胞超微结构的变化,并用TUNEL法检测睾丸中生精细胞凋亡的变化,苏丹Ⅲ染色法检测睾丸生精小管内脂滴分布的变化。结果显示:NO的生成与DES呈剂量依赖性。DES处理后,在1mg/kg体重剂量组,大量生精细胞表达eNOS和iNOS,并出现大量凋亡,退化的生精细胞胞浆内有大量髓样结构,并有大量脂滴分布于生精细胞内和细胞间。eNOS和iNOS阳性生精细胞与凋亡的生精细胞数量和类型基本一致,主要为精母细胞和圆形精子细胞。  相似文献   

4.
睾丸是雄性动物的生殖器官,具有产生雄激素和雄性生殖细胞的功能。因此,睾丸的发育形态对猪养殖生产具有重大影响。本实验选取DLY公猪胚胎期(妊娠105 d)和成年期(日龄200 d)睾丸为研究对象,对睾丸组织进行形态学观察和高通量转录组测序,并对差异表达基因进行功能富集分析。形态学观察结果表明:相较于雄性胎猪,成年公猪生精小管直径更大,生精上皮更厚,已有完整的生精结构,并可见各发育阶段精子细胞。转录组测序结果表明,共鉴定出6 296个差异表达基因。差异基因显著富集在精子发生、多细胞生物发育和细胞分化等生物过程,可能在代谢、细胞黏附、细胞周期和AMPK等通路发挥作用。本研究结果揭示了公猪胚胎期和成年期睾丸组织的形态学和转录组学的差异,为后期猪睾丸发育研究提供参考。  相似文献   

5.
研究糖尿病对小鼠睾丸的组织学影响,检测分析Ghrelin在糖尿病小鼠睾丸内的表达变化特点。通过腹腔注射四氧嘧啶的方法建立糖尿病小鼠模型,分别于造模后第5、10、15天取睾丸组织,采用石蜡切片技术和HE染色方法观察糖尿病小鼠睾丸的组织学变化特点;采用免疫组化染色方法研究糖尿病小鼠睾丸内Ghrelin的表达和分布情况。HE染色结果显示,与对照组相比,随着糖尿病病程的延长,糖尿病小鼠的生精小管内生精细胞数量呈减少趋势,排列层数及紧凑程度呈现不同程度的下降,排列松散无序,并且可见精细胞从基膜脱落。免疫组化结果显示,Ghrelin在对照组小鼠睾丸的初级精母细胞和次级精母细胞的胞膜和胞质内均呈强阳性表达,精原细胞、精子细胞和精子内无阳性表达,支持细胞和睾丸间质细胞的细胞核呈阳性表达。Ghrelin在糖尿病小鼠睾丸内的表达和分布特点与对照组相似,但阳性表达水平较对照组明显增强;随着糖尿病病程的延长,组内Ghrelin表达没有明显变化。试验证实了糖尿病对小鼠睾丸发育和Ghrelin在睾丸内的表达水平均具有一定影响,提示糖尿病对小鼠的生精能力具有一定的抑制作用。  相似文献   

6.
以中华鳖2个繁殖阶段的睾丸为研究对象,探究睾丸界膜细胞成分的变化,为中华鳖界膜的深入研究提供理论依据。分别在冬眠期(4月)以及繁殖期(9月)摘取健康成熟的野生中华鳖睾丸,使用HE染色、马松染色、甲苯胺蓝染色、免疫组化染色等方法观察不同阶段中华鳖睾丸组织及细胞成分变化情况,使用免疫组化对肌样成纤维细胞进行表达定位,对中华鳖睾丸生精小管及睾丸界膜变化进行统计分析。结果显示:繁殖期中华鳖睾丸生精小管面积显著大于冬眠期(P0.05),生精上皮厚度极显著大于冬眠期(P0.01);冬眠期中华鳖睾丸界膜面积显著大于繁殖期(P0.05),界膜厚度极显著大于繁殖期(P0.01)。冬眠期细胞间质内含有大量细胞,胶原纤维增多,而繁殖期中华鳖睾丸界膜细胞成分少,胶原纤维减少;冬眠期中华鳖睾丸的肌样成纤维细胞较繁殖期有着明显的多层弥散状结构,而繁殖期中华鳖睾丸肌样成纤维细胞的分层结构并不明显。提示:睾丸结构的改变可能与中华鳖不同繁殖阶段的生理功能相关,这也为研究界膜成分的变化对于生精障碍提供了一定的参考。  相似文献   

7.
探讨不同时期山羊睾丸组织中干细胞的分布和特点,以45 d和75 d山羊胎儿及出生后4月龄山羊睾丸为试验材料,进行了HE染色和Oct4抗体的免疫组化染色。结果显示,在45 d山羊胎儿睾丸组织中睾丸实质细胞聚集呈团状,形成岛屿状组织结构;发育到75 d山羊胎儿睾丸组织中睾丸的间质与实质组织结构差异明显,睾丸生精小管已经形成,生精小管管腔闭锁;出生后4月龄山羊睾丸组织结构明显,生精小管形态特征清晰,但管腔仍处于闭锁状态;Oct4免疫组织化学显示在45 d山羊胎儿睾丸组织中Oct4阳性细胞呈岛屿状分布,多见于生精小管前期的组成细胞;对于75 d山羊胎儿睾丸组织Oct4阳性细胞呈环状分布于生精小管中,睾丸间质组成细胞中则较少;在出生后4月龄山羊睾丸中阳性细胞主要分布于睾丸组织生精小管基底膜,沿基底膜环状分布。试验证实了在早期山羊睾丸组织中Oct4主要表达在生精前体细胞。  相似文献   

8.
制作睾丸组织切片,运用HE染色和细胞TUNEL染色分别观察不同发育阶段猪睾丸的形态学和细胞凋亡;提取睾丸组织蛋白,利用免疫印迹检测不同发育阶段猪睾丸蛋白的Kcr修饰水平;最后利用免疫荧光技术检测Kcr蛋白在猪睾丸组织中的定位.结果 显示,随着月龄的增长,猪睾丸精细管管腔直径增加,睾丸组织在4月龄时精细胞开始分裂,凋亡细...  相似文献   

9.
探讨高原老龄牦牛睾丸组织结构特点与其生殖机能的关系。采集9~13岁健康老龄牦牛睾丸组织,应用组织化学染色和透射电镜技术,观察其细胞化学特点及结构特征。结果显示,老龄牦牛睾丸体积及重量左侧略大于右侧,光镜下睾丸被膜、间质组织胶原纤维及网状纤维丰富,小叶不明显,Leydig细胞散在于结缔组织之间,生精小管不同程度退化,生精上皮层数为2~5层,细胞间隙变大,部分脱落,Sertoli细胞低矮,细胞质较少。电镜观察Sertoli细胞核外形不规则,内质网呈疏松的泡状,次级溶酶体增加,体积增大。相邻Sertoli细胞间没有观察到并行的内质网层和细丝束这一血-睾屏障所特有外质特化区形态结构,部分生精小管固有膜胶原纤维增生,上皮皱缩;Leydig细胞内脂滴异常丰富,毛细血管内皮细胞间存在紧密连接。间质组织偶有肥大细胞,间质血管及生精小管固有膜中PAS及AB-PAS阳性反应明显。老龄牦牛Sertoli细胞细胞器结构异常,血-睾屏障中Sertoli细胞间的连接处缺陷,且Sertoli细胞与基膜连接的改变可能影响了Sertoli细胞和生精细胞之间的相互作用,进而抑制了生精细胞的功能。  相似文献   

10.
为进一步深入研究CREM基因在雄性动物中的表达调控机制,试验成功建立体外生精细胞和支持细胞共培养体系,并应用免疫细胞化学研究CREM在体外培养山羊睾丸生精细胞的表达特点。结果显示:CREM仅表达于圆形精子细胞,其他生精细胞和支持细胞检测不到CREM的表达。表明CREM基因在圆形精子细胞发育过程中起重要调节作用,但其作用机制有待深入研究。  相似文献   

11.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

12.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

13.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

14.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

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16.
An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.  相似文献   

17.
Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein  相似文献   

18.
A knowledge of the microbiological status of milk and of the different structures in the mammary glands has great importance in elucidating the pathogenesis of mammary gland infections. The objective of this study was to evaluate the microbiological status of various structures in the mammary glands from naturally infected dairy cows following slaughter. A total of 94 samples of milk, 184 samples of mammary parenchyma, 168 samples of gland cisterns, and 168 samples of teat cisterns were collected for microbiological examination. Microorganisms were detected in 59.9% of all samples, 67.0% of the milk samples, 70.1% of the mammary parenchymas, 55.9% of the gland cisterns and 48.8% of the teat cistern samples. When all samples were considered, coagulase-negative Staphylococcus were the most prevalent (35.7%) followed by coagulase-positive Staphylococcus (12.2%), Corynebacterium bovis (2.4%), Prototheca sp. (1.9%), and Streptococcus dysgalactiae (1.5%). There was a significantly higher occurrence of microorganisms in the milk and mammary parenchyma compared to the gland cisterns and teat cisterns. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

19.
Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L3) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L3 developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.  相似文献   

20.
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