Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

B cell differentiation in the bursa of Fabricius and spleen of embryos and chicks immediately after hatching

Flow cytometric analysis and immunohistochemical observation were used to qualitatively and quantitatively clarify the nature of B cell differentiation in the bursa of Fabricius of chick embryos and to determine the timing of antibody class switching in chicken spleens based on positivity of IgM and IgG on and in the cells. In the bursa, the sIgM‐positive cell population formed from the 12^th to 15^th day of embryogenesis. The proportion of sIgM‐high expressing (sIgM^high) cells was lower among bursacytes than splenocytes of hatched chicks, suggesting that the sIgM^high bursacytes are to be released to peripheral sites. The proportion of sIgM^high cells was higher at 0 days old than at any other examined stage of development. Colonization of the spleen by B cells occurred between the 18^th day of embryogenesis and 0 days old. Antibody class switching was thought to start in the spleen between 1 and 2 weeks of age, because IgG‐positive cells were present in the spleen of 2‐week‐old chicks, but not 0‐day‐old or 1‐week‐old chicks.     

神经肽Y及其Y1受体阳性细胞在鸭腔上囊中的分布及发育变化(英文)

神经肽Y及其Y1受体在神经-免疫-网路中发挥着重要作用。二者在许多脊椎动物胸腺和脾脏中的表达已有研究,但它们在腔上囊中的表达及发育变化特征还未见报道。本试验应用免疫组化技术研究神经肽Y及其Y1受体阳性细胞在鸭腔上囊胚胎及胚后发育中的分布及变化特征。结果显示,在鸭腔上囊中神经肽Y及其Y1受体分别最早表达于26和22d胚龄。神经肽Y阳性细胞分布于黏膜上皮、滤泡、肌层和小动脉平滑肌。Y1受体阳性细胞分布于黏膜上皮、滤泡淋巴细胞。各部位神经肽Y及其Y1受体阳性细胞数量表现出明显增龄变化特征。结果说明鸭腔上囊中存在神经肽Y及其Y1受体,二者可能存在的相互作用主要表现在胚后发育阶段,这与B淋巴细胞的发育与分化以及腔上囊的退化密切相关。     

Immunolocalization of steroidogenic enzymes and their expression during the breeding season in the testes of wild raccoon dogs (Nyctereutes procyonoides)

The objective of this study was to investigate immunolocalization of steroidogenic enzymes 3βHSD, P450c17 and P450arom and their expression during the breeding season in wild male raccoon dogs. The testicular weight, size and seminiferous tubule diameters were measured, and histological and immunohistochemical observations of testes were performed. The messenger RNA expression (mRNA) of 3βHSD, P450c17 and P450arom was measured in the testes during the breeding season. 3βHSD was found in Leydig cells during the breeding and non‐breeding seasons with more intense staining in the breeding season. P450c17 was identified in Leydig cells and spermatids in the breeding season, whereas it was present only in Leydig cells in the non‐breeding season. The localization of P450arom changed seasonally: no immunostaining in the non‐breeding season; more extensive immunostaining in Leydig cells, Sertoli cells and elongating spermatids in the breeding season. In addition, 3βHSD, P450c17 and P450arom mRNA were also expressed in the testes during the breeding season. These results suggested that seasonal changes in testicular weight, size and seminiferous tubule diameter in the wild raccoon dog were correlated with spermatogenesis and immunoreactivity of steroidogenic enzymes and that steroidogenic enzymes may play an important role in the spermatogenesis and testicular recrudescence and regression process.     

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.     

Effect of growth hormone on estrogen receptor binding properties in the chick oviduct in vivo

The equilibrium dissociation constant (K_d) and the maximum binding capacity (B_max) of estrogen receptor in cytosolic and nuclear fractions in oviduct of pullets following various daily injections (1–10 times) of growth hormone (GH: 50 µg/chick) were examined by Scathchard analysis of specific [³H]estradiol‐17β (E₂) binding. The K_d values of receptor in both fractions decreased after three times of GH‐injection. In the B_max values, three times of the injection caused a marked decrease in that value in the cytosolic fraction with a concomitant increase in that in the nuclear fraction, whereas the total B_max (sum of B_max in the cytosolic and nuclear fractions) did not change. A similar relationship between the K_d values in the binding of the two fractions was also observed in 4–10 times of GH‐injection. However, in 4–10 times of GH‐injection the B_max of the both fractions and total B_max was greater than that in vehicle‐injection (control). When the chicks were injected with 6–10 times of GH‐injection, the weight of oviduct was increased. No change in the plasma concentrations of E₂ was found following GH‐ and vehicle‐injections into the chicks. The results suggest that growth hormone stimulates the growth of the chick oviduct by increasing the binding affinity and capacity of estrogen receptor.     

鸡包涵体肝炎鸡胚肝细胞包涵体特性的研究

应用鸡包涵体肝炎病毒 FAV- Hb株试验感染 SPF鸡胚 ,通过透射电镜对感染鸡胚肝脏的观察 ,表明 FAV- Hb株可引起鸡胚肝细胞内形成三种类型的包涵体 ,即非病毒性中等电子密度包涵体 ,病毒性包涵体和非病毒性高电子密度包涵体 ,各型包涵体均可见于胞核或胞浆内 ,但胞核是包涵体首先形成的部位 ,核内包涵体通过核膜进入胞浆。各型包涵体在其形成过程中有较密切的关系。     

Expression of estrogen receptor 1 and progesterone receptor in primary goat mammary epithelial cells

The exact role and sensitivity of cells to estrogen and progesterone mediated through the steroid receptors during lactation is not known. Expression of estrogen receptor 1 (ESR1) and progesterone receptor (PGR) was quantified in mammary tissue‐derived primary goat mammary epithelial cells (pgMECs) to determine the influence of donor tissue physiology (lactating and juvenile) and cell culture growth conditions (basal and lactogenic) on ESR1 and PGR expression in the derived cells. Relative messenger RNA (mRNA) levels for both receptors were the highest in cell lines derived from mammary tissue of juvenile goats. Maintaining pgMECs in lactogenic conditions resulted in up‐regulation of ESR1 (1.36‐ to 12.35‐fold) and in down‐regulation of PGR (‐2.53‐ to ‐3.62‐fold), compared to basal conditions. Based on Western blotting analysis we suggest that the differences in mRNA expression are translated to the protein level. We suggest that differential expression in lactating conditions is correlated with terminal differentiation of the pgMECs. Double immunostainings showed that estrogen receptor alpha (ER‐α) positive cells do not exclusively belong to the luminal lineage and that ER‐α and PGR can be expressed individually or co‐expressed in the pgMECs. The derived primary cultures/lines in early passages are hormone‐responsive and represent a useful surrogate for mammary tissue in research experiments.     

Kinetics of phosphorus absorption and expressions of related transporters in primary cultured duodenal epithelial cells of chick embryos

Two experiments were conducted to investigate the kinetics of phosphorus (P) absorption and expressions of type IIb sodium-dependent phosphate cotransporter (NaP-IIb), inorganic phosphate transporters 1 and 2 (PiT-1 and PiT-2) in primary cultured duodenal epithelial cells of chick embryos. In experiment 1, the P absorptions across duodenal epithelial cell monolayers at different incubation time points (0, 10, 20, 40, 60, 80, 100 and 120 min) were compared. In experiment 2, the kinetics of P absorption was performed at 40 min after incubation of duodenal epithelial cells with the media containing 0, 0.75, 1.5, 3.0, 6.0, 12.0, 24.0 and 48.0 mmol P/L as KH₂PO₄, and the mRNA and protein expression levels of NaP-IIb, PiT-1 and PiT-2 in duodenal epithelial cells with the media containing 0, 6.0 and 48.0 mmol P/L were determined at 87 min after incubation. The results from experiment 1 showed that the P absorption increased linearly (p < .0001) from 0 to 80 min and the fastest increase occurred at 40 min; the asymptotic model was shown to have the best fit degree, and the optimal incubation time for saturable P absorption was determined to be 87 min. The kinetic curves of P absorption from experiment 2 demonstrated that P absorption was a mixed process of a non-saturable diffusion plus a saturable carrier-mediated transport across the duodenal epithelial cells. The high P concentration (48.0 mmol/L) decreased (p < .05) NaP-IIb and PiT-1 mRNA and protein levels and increased (p < .0001) PiT-2 mRNA level. These results indicated that the P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos was a mixed process of a non-saturable diffusion plus a saturable carrier-mediated transport and could be restricted by reducing the NaP-IIb and PiT-1 expressions while increasing the PiT-2 expression at a high P concentration.     

催情助孕液对小鼠雌、孕激素水平及其受体mRNA表达的影响

分别以0.1、0.2、0.4 mL/d剂量的催情助孕液给21日龄小鼠灌服7d后,检测体质量、脏器指数、血清雌激素和孕激素水平、子宫组织中雌激素和孕激素受体基因mRNA的表达情况,并与腹腔注射雌二醇4d的小鼠进行比较.结果显示与对照组和雌二醇注射组比较,0.2 mL的催情助孕液能显著促进小鼠子宫发育,并增强雌激素水平及其受体基因表达,而对孕激素水平及其受体基因表达无显著影响.结果表明,催情助孕液对动物生殖发育与促进发情具有较好的效果.     

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。     

IBDV感染对法氏囊RNA m~6A和DNA 5mC修饰以及相关催化酶基因表达的影响

传染性法氏囊病毒(IBDV)感染造成病鸡法氏囊严重损伤,引起免疫抑制,然其具体机制仍不明晰。本研究旨在探讨IBDV感染对法氏囊表遗传修饰及其相关酶表达的影响。试验采用21日龄商品肉鸡,给予IBDV处理,感染后3,5 d收集法氏囊组织。应用基于ELISA试剂盒检测总5mC和m~6A,Real-time PCR检测相关催化酶基因表达。结果显示,感染后5 d,总m~6A水平及其甲基转移酶METTL3和METTL14 mRNA表达均显著降低(P0.01);去甲基化酶FTO的表达在感染后3,5 d均显著降低(P0.05)。总5mC水平在感染后3 d明显升高(P0.05),其去甲基化酶Tet1 mRNA表达明显降低(P0.01);感染后5 d,甲基转移酶DNMT1 mRNA表达显著降低(P0.01),而DNMT3a的表达明显升高(P0.01);结果表明IBDV能够影响法氏囊5mC和m~6A谱及相关酶的表达,这可能是IBDV影响宿主免疫功能的途径之一。     

雌激素对大鼠小脑中Bcl-2和Bax蛋白表达的影响

采用免疫组织化学超敏SP法检测摘除卵巢及用外源性17β-雌二醇治疗后SD大鼠小脑中Bcl-2和Bax蛋白表达的变化.结果显示,摘除双侧卵巢后,SD大鼠小脑皮质及深层核团中Bcl-2的表达有不同程度的降低,而Bax的表达出现不同程度的上升;给予外源性17β-雌二醇治疗后,2种蛋白的比例基本恢复正常.表明,雌激素能够下调小脑皮层及小脑深层核团中Bax蛋白的表达,上调Bcl-2蛋白的表达,对小脑神经元具有保护作用.     

Molecular expression and identification of caprine estrogen receptor gene 1 for fertility status in bucks

Estrogen and its receptors are essential for sexual development and reproduction. Oestrogen receptor alpha (ERα) is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene ESR1 (oestrogen receptor1) responsible for better fertility. ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. The present study was undertaken to investigate the expression and presence of ESR1 gene in fertile and low-fertile male goat breeds. We identified ESR1 gene through various molecular tools. Genotyping was carried out by high resonance melting analysis using Roche Light Cycler 480(LC-480) system and found three different genotypes. Genotypic frequency-AA (blue-0.67), BB(Red-0.2), AB(Green-0.08) with allele frequency A(0.71 and B (0.29). The predominance of this gene in head of epididymis in fertile bucks was confirmed by SDS-PAGE, Western blotting and immunohistochemistry. From the results, we corroborated that the present study provides a useful and effective way to predict male fertility in goat breeds, which in turn increases the percentage of fertility in flock leading to more number of offspring in a kidding season.     

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.