降钙素基因相关肽在鸭腔上囊胚胎及胚后发育过程中的表达

应用免疫组织化学技术研究降钙素基因相关肽在鸭腔上囊胚胎及胚后发育过程中的表达特征.结果显示,降钙素基因相关肽强阳性细胞出现于20 d胚龄至胚后14周龄的滤泡间上皮,20 d胚龄至胚后17周龄的小结相关上皮,新生雏至胚后14周龄的滤泡皮质,24 d胚龄至胚后14周龄的滤泡髓质,20 d胚龄至胚后14周龄的肌层和小动脉平滑肌.结果说明,降钙素基因相关肽表达于鸭腔上囊的不同部位及不同发育阶段;在腔上囊退化期,降钙素基因相关肽的表达表现出明显的变化特征,这可能是导致腔上囊退化的原因之一;滤泡间上皮与小结相关上皮在神经肽表达方面存在差异.     

5种弥散神经内分泌细胞在鸭腔上囊中的表达特征（英文）

应用免疫组化技术并结合图像分析软件研究了胃泌素、β-内啡肽、胰高血糖素、5-羟色胺、生长抑素在鸭腔上囊中的表达特征。结果显示：鸭腔上囊小结相关上皮细胞呈胃泌素强阳性反应;滤泡间粘膜上皮呈β-内啡肽中强度阳性反应;淋巴滤泡皮质部有少量的β-内啡肽、胃泌素、5-羟色胺阳性细胞,呈中、强度免疫反应;淋巴滤泡髓质有胃泌素强阳性细胞;生长抑素和胰高血糖素均呈阴性反应。结果说明,鸭腔上囊中存在β-内啡肽、胃泌素、5-羟色胺弥散神经内分泌细胞,它们在淋巴滤泡皮质部的表达特征有利于其通过内分泌和旁分泌方式调节B细胞的发育。     

5种弥散神经内分泌细胞在鸭腔上囊中的表达特征

应用免疫组化技术并结合图像分析软件研究了胃泌素、β-内啡肽、胰高血糖素、5-羟色胺、生长押素在鸭腔上囊中的表达特征.结果显示鸭腔上囊小结相关上皮细胞呈胃泌素强阳性反应;滤泡间粘膜上皮呈β-内啡肽中强度阳性反应;淋巴滤泡皮质部有少量的β-内啡肽、胃泌素、5-羟色胺阳性细胞,呈中、强度免疫反应;淋巴滤泡髓质有胃泌素强阳性细胞;生长抑素和胰高血糖素均呈阴性反应.结果说明,鸭腔上囊中存在β-内啡肤、胃泌素、5-羟色胺弥散神经内分泌细胞,它们在淋巴滤泡皮质部的表达特征有利于其通过内分泌和旁分泌方式调节B细胞的发育.     

胚胎及胚后发育期鸭腔上囊淋巴细胞增殖与凋亡的研究

采用PCNA免疫组化法和TUNEL染色法,并结合光、电镜技术研究天府肉鸭胚胎及胚后发育期腔上囊淋巴细胞增殖与凋亡的动态变化规律。结果显示胚胎期腔上囊淋巴细胞增殖明显,其滤泡淋巴细胞增殖指数（PIF）随胚龄增加而逐渐增高,26d胚龄达峰值。胚后期各组滤泡皮质淋巴细胞增殖指数（PIC）和髓质淋巴细胞增殖指数（PIM）均呈下降趋势;各组PIM均明显高于PIC。胚胎期腔上囊滤泡淋巴细胞凋亡指数（AIF）随胚龄增大而逐渐增高。胚后期O～3周龄滤泡髓质淋巴细胞凋亡指数（AIM）继续增高,滤泡皮质淋巴细胞凋亡指数（AIC）则无明显变化;AIC和AIM在5周龄下降,17周龄明显升高,29周龄达峰值。胚后0～14周龄,各组AIM均明显高、于AIC,而17～29周龄AIM则明显低于AIC。凋亡淋巴细胞核呈现多种形态,线粒体肿胀,嵴断裂。结果提示淋巴细胞增殖与凋亡在鸭腔上囊胚胎及胚后发育的整个过程中普遍存在,具有明显增龄变化特性,二者协同参与腔上囊发育和退化过程。     

雏鸭腔上囊的生长及组织发育

通过对1～49日龄雏鸭腔上囊绝对质量和生长指数测定以及组织学观察,探讨雏鸭腔上囊生长及组织发育规律。结果:随日龄增长,腔上囊绝对质量逐渐增高;腔上囊生长指数21日龄达最高;黏膜大皱褶高、宽度、淋巴滤泡面积、皮质宽度均不断增加;小结相关上皮向腔面形成突起,并不断增高增宽。结果表明,雏鸭腔上囊1～14日龄生长较为缓慢,14～35日龄生长较快,35～49日龄发育基本趋于稳定,其中21～28日龄是雏鸭腔上囊生长发育的高峰时期,28日龄时,雏鸭腔上囊组织结构基本发育成熟。     

神经肽Y阳性神经元在皖西白鹅小肠的分布

用免疫组化SABC法,观察神经肽Y(NPY)免疫反应阳性物质在成年皖西白鹅小肠的分布。结果表明神经肽Y阳性神经元主要分布于皖西白鹅的十二指肠、空肠和回肠的黏膜、黏膜下组织。细胞体形态多样,数量从十二指肠向空肠和回肠逐渐减少,密度分别为(62.10±4.25)/mm2、(51.14±2.12)/mm2、和(41.21±3.15)/mm2。神经纤维主要分布于小肠肌层。本文还讨论了神经肽Y的起源及对小肠黏膜功能的调节作用。     

鸡回肠中T淋巴细胞及其亚群发育的试验

为探寻鸡回肠中T淋巴细胞及其亚群的发育规律,本试验通过免疫组织化学方法,应用CD3、CD4和CD8单克隆抗体研究鸡回肠中T淋巴细胞及其亚群出现、迁移、定位分布及数量变化过程.结果显示,CD3+、CD8+T淋巴细胞最初于18胚龄时出现,CD4+T淋巴细胞于出壳后1日龄时出现.在定位分布上,CD3+细胞在黏膜上皮内以及固有层中均匀分布,CD4+细胞以固有层中的分布为主,黏膜上皮内的分布较少.CD8+细胞最初主要分布在黏膜固有层中;随后,CD8+细胞逐渐向上皮内迁移;最终,黏膜上皮内出现广泛的CD8+细胞浸润.在数量变化上,CD3+、CD4+及CD8+整体呈逐渐增加趋势,第2周时阳性细胞数量稍有下降,21日龄时显著增加到达较高水平后保持稳定.结果表明,鸡出壳后,回肠的细胞免疫功能逐渐增强,并在21日龄时到达成熟水平.     

神经内分泌细胞在鸭胸腺中的表达特征

应用免疫组化技术并结合图像分析软件研究了β-内啡肽、胃泌素、胰高血糖素、5-羟色胺、血管活性肠肽、神经肽Y、生长抑素细胞在鸭胸腺中的表达特征。结果显示：7种神经内分泌细胞在胸腺中呈强阳性表达;β-内啡肽阳性细胞在皮质部较多,而胃泌素、胰高血糖素、5-羟色胺、血管活性肠肽、神经肽Y、生长抑素阳性细胞在髓质和皮髓质交界区的数量高于皮质部;除胰高血糖素外,其余6种神经内分泌激素在胸腺小体中呈不同程度的阳性反应。本试验结果说明,鸭胸腺不仅是中枢免疫器官,而且具有重要的神经内分泌功能;β-内啡肽阳性细胞在T淋巴细胞的发育中发挥重要作用;胃泌素、胰高血糖素、5-羟色胺、生长抑素、血管活性肠肽、神经肽Y阳性细胞在髓质部和皮髓质交界区的表达有利于其通过内分泌、自分泌或旁分泌方式调节T细胞的发育。同时,也探讨了胸腺小体的功能。     

鸭腔上囊胚胎及胚后发育期凋亡相关蛋白的免疫组化分析

研究天府肉鸭腔上囊胚胎及胚后发育期Bcl-2、Bax、Fas、FasL蛋白的表达。将20只天府肉鸭分为4组,即24天胚龄(E24),胚后3、8、29周龄(P3、P8、P29),采用免疫组化技术。结果显示,Bcl-2、Bax、Fas、FasL在各组滤泡淋巴细胞中均有表达,FasL还表达于滤泡间上皮。滤泡皮质和髓质淋巴细胞Bcl-2阳性率呈递减的变化趋势(髓质P3～8除外);皮质和髓质淋巴细胞Bax阳性率在E24～P8恒定,P29上升;皮质和髓质淋巴细胞Bcl-2/Bax值呈下降趋势,但皮质在P3～8恒定,髓质在P3～8上升;滤泡皮质和髓质淋巴细胞Fas阳性率在E24～P3上升,P3～8下降,P8～29上升;皮质和髓质淋巴细胞FasL阳性率变化规律与Fas相似,但皮质在P3～8恒定。滤泡皮质淋巴细胞Bcl-2阳性率及Bcl-2/Bax值在E24～P8明显高于髓质,在P29则显著低于髓质;皮质淋巴细胞Bax阳性率在E24～P8与髓质无显著差异,在P29则明显高于髓质;皮质淋巴细胞Fas和FasL阳性率在E24～P8明显低于髓质,在P29则显著高于髓质。结果提示,Bcl-2、Bax、Fas、FasL是鸭腔上囊胚胎及胚...     

鸡十二指肠中T淋巴细胞及其亚群发育的研究

通过免疫组织化学法,应用CD3、CD4和CD8单克隆抗体研究了CD3+T淋巴细胞及CD4+和CD8+T淋巴细胞亚群在鸡盲肠扁桃体中的出现、迁移、组织定位分布及数量变化规律等一系列发育过程。结果显示:CD3+、CD8+T淋巴细胞最初于11胚龄时出现,而CD4+T淋巴细胞在15胚龄时出现。在定位分布变化上,CD3+主要分布在黏膜上皮中,CD4+主要分布在黏膜固有层中,CD8+在黏膜固有层和黏膜上皮内都有大量分布。在数量变化上,1～7日龄雏鸡各种阳性细胞的数量都骤然增加;而到7日龄时,雏鸡CD3+淋巴细胞的数量明显减少,CD4+、CD8+T细胞的数量无明显变化;从21日龄开始直到35日龄,雏鸡CD3+、CD4+和CD8+T淋巴细胞的数量持续增加。试验证明,鸡在出壳后初期十二指肠的细胞免疫功能增强。     

Ontogeny expression of ghrelin,neuropeptide Y and cholecystokinin in blunt snout bream,Megalobrama amblycephala

Ghrelin, neuropeptide Y (NPY) and cholecystokinin (CCK) all have important roles in the regulation of feeding in fish and mammals. To better understand the role of the three peptides in appetite regulation in the early developmental stages of blunt snout bream (Megalobrama amblycephala), partial cDNA sequences of ghrelin, NPY and CCK genes were cloned. And then, real‐time quantitative PCR and RT‐PCR were used to detect and quantify the mRNA expressions of these genes from zygotes to larvae of 50 days after hatching (DAH). Ghrelin, NPY and CCK were all expressed throughout the embryonic and larval development stages, and the expression levels were higher in larval stages than in embryonic stages. Ghrelin and NPY mRNA expressions were upregulated at 1, 3, 5 DAH, while CCK mRNA expression was reduced significantly at 3 DAH. The mRNA expression levels of three genes in larvae varied significantly until 30 DAH. In adult fish, all three peptides were detected to be expressed in brain and several peripheral tissues. Ghrelin mRNA was mainly expressed in the intestine, whereas NPY and CCK mRNAs were mainly expressed in the brain. Taken together, these results indicate that ghrelin, NPY and CCK may have roles in early development and participate in the regulation of feeding of larvae in blunt snout bream and will be helpful for further investigation into feed intake regulation in adults of this species.     

Histological evaluation of immune organs in chicken embryos inoculated with Marek's disease virus and lymphokines

The aim of the present study was to evaluate the presence of lymphocytes and granulocytes in different stages of embryonic development and on the first posthatching day. The lymphocytes present in the bursa of Fabricius and thymus were evaluated by histological analysis of the yolk sac, bursa of Fabricius, thymus, liver and bone marrow of 100 chicken embryos divided into groups and treated with: (I) Marek's disease vaccine as viral antigen, (II) Marek's disease vaccine plus lymphokines, (III) lymphokines, and (IV) vaccine diluent. Group V was not treated. Samples were taken on days 14, 17 and 20 of incubation and on the first posthatching day. An increase in the number of epithelial matrix as precursors of lymphoid follicles was observed in the bursa of Fabricius of embryos inoculated with lymphokines compared to embryos in all the other groups (p < 0.05). In addition, a higher amount of granulocytes was found in the yolk sac and liver of embryos inoculated with lymphokines than in the embryos of all other groups (p < 0.05). In the bone marrow, no significant difference was observed among the treated groups concerning the amount of granulocytes. The results suggest that administration of antigens or protein molecules at an early stage of embryonic development increases the presence of granulocytes in the liver and granulopoiesis in the yolk sac, and also increases the number of epithelial matrixs in the bursa of Fabricius.     

Developmental changes in cell proliferation and apoptosis in the normal duck bursa of Fabricius

The aim of this work was to investigate developmental changes in cell proliferation and apoptosis in normal duck bursa of Fabricius using flow cytometry and immunohistochemistry. Studies were carried out on Tianfu ducks on days 24 and 27 of embryogenesis (E24 and E27) along with days 20, 70, and 200 of postnatal development (P20, P70, and P200). Results showed that the percentage of G₀/G₁ bursa cells significantly increased between E24 and P200 while the percentage of cells in the S phase or G₂ + M phase as well as the proliferating index obviously decreased during the same period. Proliferation cell nuclear antigen was detected in lymphocyte and interfollicular epithelium. The proliferative lymphocyte density tended to decrease from E24 to P200. Apoptotic bodies in macrophages, free apoptotic bodies, or nuclei with condensed chromatin in lymphocytes in follicles were identified by transferase-mediated dUTP nick-end labeling. Both flow cytometry and microscopic analysis reveal that the proportion of apoptotic cells and apoptotic lymphocyte density increased from E24 to P20, fell on P70, then rose again on P200. Our foundings demonstrate that cell proliferation decreases and apoptosis increases with age. These changes may account for duck bursa development and involution.     

谷氨酰胺对肉鸡免疫器官胚后发育的影响

160只1日龄艾维茵肉仔鸡随机分成4组。分别饲喂添加0％、0．2％、0．4％和0．8％谷氨酰胺（Gln）饲粮28d。每周末取鸡24只，每组6只，颈静脉放血致死，取胸腺、法氏囊和脾脏，称重．制作切片．光镜观察，研究基础日粮中添加Gln对肉鸡免疫器官胚后发育的影响。结果显示：（1）添加0．8％Gln组1、4周龄和添加0．4％Gln组2、4周龄胸腺重量显著高于对照组（P〈0．05），添加0．8％Gln组2周龄极显著高于对照组（P〈0．01）；添加0．4％Gln组1、2、4周龄和添加0．8％Gln组4周龄法氏囊重量显著高于对照组（P〈0．05），添加0．8％Gln组1、2周龄极显著高于对照组（P〈0．01）；试验各组脾脏重量均高于对照组。添加0．8％Gln组4周龄显著高于对照组（P〈0．05）。（2）试验组胸腺小叶皮质增厚，淋巴细胞密集，髓质比例减小。胸腺小体减少、减小。试验组法氏囊皱襞内淋巴滤泡增多，皮质增厚．淋巴细胞增多。试验组脾脏脾小结增多、增大；动脉周围淋巴鞘增厚；椭球增多、增大．细胞排列疏松．鞘毛细血管管腔变大．内皮细胞增高。间隙增大。添加0．8％Gln对肉鸡免疫器官发育影响最为明显。结果表明，日粮中添加Gln对肉鸡中枢淋巴器官胚后发育的促进作用优于外周淋巴器官。并有延缓法氏囊退化作用。本试验从组织学角度证明Gln能够促进机体淋巴细胞的增殖分化及免疫器官的胚后发育，进而提高机体的免疫功能。     

Effects of estrogen on estrogen receptor expression in the bursal cells of chick embryos and steroidogenic enzymes gene expression in the bursa: Relevance of estrogen receptor and estrogen synthesis in the bursa of chick embryos

Estrogen has been reported to act on B cell genesis in the bursa of Fabricius of chick embryos. In this study, we attempted to demonstrate the hypothesis that B cell genesis is controlled by estrogen receptor (ER) in the bursal cells and steroidogenic enzymes synthesized in the bursa. We previously reported the presence of estrogen receptor α (ERα) in the bursa during the late stage of embryogenesis and an increase in the expression of ERα messenger RNA (mRNA) between the 13th day and 16th day. The number of ER-positive cells was maximal on the 16th day. In the present study, ER-positive cells in the bursa during the late stage of embryogenesis increased 4 h after β-estradiol treatment on the 14th to 18th day. The concentration of β-estradiol in the embryonic bursa increased. These results suggest that this stage of embryogenesis is critical in B cell development in the bursa in connection with the effect of estrogen treatment. Our findings also showed that the mRNA expression of five steroidogenic enzymes occurred in the bursa of chick embryos. These results suggest that estrogen is synthesized in the embryonic bursa and estrogen acts on the bursal cells in a paracrine fashion.     

Embryotoxicity assay of aflatoxin produced by Aspergillus parasiticus NRRL 2999

1. The embryotoxicity of mixed aflatoxins (AF) and aflatoxin B1 (AFB1) were evaluated by a modified chick embryotoxicity screening test (CHEST). Adverse effects on the early embryonic development of thymus and bursa of Fabricius were also investigated by light microscopy. AF consisted of 83.06% AFB1, 12.98% AFB2, 2.84% AFG1 and 1.12% AFG2. 2. A total of 448 fertilised laying hens' eggs were used. AF and AFB1 were injected into the eggs at doses of 10, 100 and 1000 ng/egg. Embryonic developmental stages were evaluated according to the Hamburger-Hamilton scale (HH-scale). 3. The results showed that AFB1 given at 10 ng/egg had a significantly (P<0.05) greater embryotoxic effect than AF given at a similar dose. The higher doses of both AF and AFB1 caused higher embryonic mortality and also an increase in early deaths. 4. In the groups receiving 100 ng/egg AF and AFB1 an abnormal development was seen, with a protruded central region, corresponding to the area pellucida of the blastoderm. No other developmental abnormality attributable to AF or AFB1 was found.     

Effects of age and strain on the expression of leptin receptor, neuropeptide Y and pro-opiomelanocortin in the hypothalamus of young chickens

1. An experiment was conducted to study changes in the expression of the hypothalamic leptin receptor, neuropeptide Y (NPY) and proopiomelanocortin (POMC) with age during the early neonatal period in two different strains of chickens: Beijing-You (BY) and Arbor Acres (AA). 2. Compared with BY chickens, AA chickens ate more, and grew faster. Hypothalamic NPY concentrations of both strains increased with age until d 7 followed by a decline. Hypothalamic NPY of BY chickens on d 7 was lower than in AA chickens at the same age. 3. No difference with age was observed in hypothalamic α-melanocyte-stimulating hormone (α-MSH) of BY chickens, while hypothalamic α-MSH in AA chickens on d 0 was higher than on the other days. Compared with AA chickens, BY chickens showed lower hypothalamic α-MSH on d 0. 4. Similar developmental changes between two strains were observed in the expression of leptin receptor, NPY or POMC genes in the hypothalamus. Hypothalamic mRNA of leptin receptor on d 0 was higher than on d 1 and 7. Unlike NPY, hypothalamic NPY mRNA on d 0 was higher than on the other days. 5. Hypothalamic POMC mRNA decreased gradually with age until 7 d followed by a slight increase. 6. The results showed that the developmental changes of hypothalamic signal molecules varied with age and strain. NPY, α-MSH and leptin receptor might be involved in the early programming of feed intake in newly hatched chickens.