β-葡糖醛酸酶在恒河猴组织中的分布

目的　阐明β -葡糖醛酸酶 (β-G)在正常恒河猴组织中的分布。方法　对实验恒河猴主要组织的冰冻切片采用后偶合技术进行了染色和显微观察。结果　肾上腺皮质束状带和网状带细胞 ,肝脏、睾丸、卵巢、胃肠道的实质细胞、枯否细胞、软骨细胞、卵巢间质腺细胞等富含β -G而深染蓝色。肾上腺皮质球状带细胞、血细胞、脑、肌肉、胆囊和胰腺的实质细胞等酶含量少而染色较弱。软骨基质 ,小脑皮质分子层和颗粒层等部位酶呈阴性。结论　动物的种系和品种不同 ,器官组织内 β-G的含量也不相同     

肾上腺皮质细胞的单层培养方法

目的: 体外单层培养大鼠肾上腺皮质细胞,以便为肾上腺皮质细胞的有关实验提供必要条件.方法: 采用胶原酶Ⅱ、透明质酸酶消化等步骤分离Wistar大鼠的肾上腺皮质细胞;分离的细胞于含15% NBS的DMEM培养基、37℃、5% CO2的环境中培养,3β-羟基类固醇脱氢酶特异染色鉴定.结果: 培养的肾上腺皮质细胞呈梭形,胞体较大、有少量突起,3β-羟基类固醇脱氢酶特异染色后胞体显著着蓝色.肾上腺皮质细胞不呈典型的"S"形增殖趋势,适于作原代培养.上皮样细胞和成纤维样细胞同时存在.结论: 通过细胞形态观察及特异酶染色鉴定,本文培养的细胞为大鼠肾上腺皮质细胞.     

雌激素对大鼠肾上腺皮质中Bcl-2和 Bax蛋白表达的影响

利用免疫组织化学SP法检测摘除卵巢及补充17β-雌二醇后大鼠肾上腺皮质各层Bcl-2和Bax蛋白表达的变化。结果表明,摘除卵巢后肾上腺皮质束状带和网状带Bcl-2和Bax表达都增加;球状带中Bax表达增加,而Bcl-2表达阴性。补充17β-雌二醇后,束状带和网状带中Bcl-2蛋白表达显著降低,Bax在束状带中表达极显著降低,而在网状带中没有阳性产物;在球状带中Bcl-2表达上升,而Bax表达下降。表明雌激素在肾上腺皮质各层发挥着复杂的作用。     

肾上腺皮质束状带特异性表达Cre重组酶转基因小鼠的构建

肾上腺是人体重要的内分泌器官。由于缺乏肾上腺皮质束状带特异性表达Cre酶的工具鼠,目前对肾上腺皮质束状带细胞中特异表达基因的功能缺乏深入的解析。CYP11B1基因编码类固醇11β-羟化酶,该酶是糖皮质激素合成的关键酶,在肾上腺皮质束状带中特异性表达。本研究旨在利用CYP11B1基因在束状带特异性表达的特点,构建在肾上腺皮质束状带中特异性表达Cre重组酶的转基因动物。采用CRISPR/Cas9技术在CYP11B1基因终止密码子位点定点敲入2A-GfpCre表达框,获得CYP11B1-2A-GfpCre同源重组载体,进而构建CYP11B1Cre小鼠,并通过mTmG和LacZ染色确定Cre酶主要表达在小鼠肾上腺皮质束状带。在此基础上,本研究还用该工具鼠与胱硫醚-γ-裂解酶(cystathionineγ-lyase, CTH)条件性敲除鼠交配,获得了肾上腺皮质束状带CTH特异性敲除的小鼠,并证实了该动物肾上腺皮质束状带中CTH表达缺失。以上结果充分说明肾上腺皮质束状带特异性表达Cre重组酶小鼠构建成功。该工具鼠的成功构建,为深入研究肾上腺皮质束状带相关功能提供了有力工具。     

日本血吸虫的组织化学研究 Ⅰ.成虫体内核酸、氨基酸、糖元和磷酸酶的分布

1.应用组织化学的方法研究了从小白鼠肠系膜静脉内取出的日本血吸虫成虫体内核酸、氨基酸、糖元和磷酸酶的分布情况。 2.成虫体内核酸的分布虽较广泛,但以雄虫的睾丸、雌虫的卵巢和卵黄腺中的含量为最高。在睾丸中主要为DNA,而卵巢和卵黄腺中则为RNA。 3.虫体的肌纤维、卵巢和睾丸中的生殖细胞核、卵黄细胞和肠管上皮细胞对显示酪氨酸、色氨酸和组氨酸的偶联重氮反应呈现阳性。角皮下肌层、实质组织细胞和卵黄细胞的颗粒滴呈强的溴酚蓝阳性反应。而对铁氰化物的反应仅卵黄细胞的颗粒滴呈现阳性。 4.糖元主要分布于虫体的实质组织和各种肌纤维内,尤以雄虫抱雌沟附近的实质组织内含量最多。在生殖器官中,仅成熟的卵细胞胞浆含有少许。 5.成虫体表角皮含有大量的碱性磷酸酶,而卵模上皮细胞、卵黄细胞和排泄管壁亦呈阳性反应。酸性磷酸酶主要分布于虫体的角皮下肌层、实质组织细胞核、肠管上皮细胞、雌雄生殖细胞和卵黄细胞内。 6.对于日本血吸虫体内各器官组织中所含的上述各种化学物质的生理意义进行了讨论,并认为血吸虫除肠道摄食外,尚可通过体表角皮吸收碳水化合物等营养物质。     

甲状腺激素受体在扬子鳄卵巢中的免疫组织化学定位和不同繁殖时期的mRNA表达水平变化检测

目的研究扬子鳄甲状腺激素受体(thyroid hormone receptor,TR)基因的系统进化关系和在卵巢组织中的定位,分析成年扬子鳄在不同繁殖活动期卵巢组织中甲状腺激素受体的动态变化规律。方法通过Clustal W软件对扬子鳄和其它脊椎动物TRs氨基酸序列进行序列比对,使用MEGA 6.0软件构建NJ系统发育树;应用q RT-PCR方法检测TRα和TRβm RNA在不同繁殖时期卵巢组织中的表达水平差异;应用免疫组织化学法对扬子鳄卵巢卵泡中TR进行组织定位。结果系统发育分析显示扬子鳄与鸟类亲缘关系较近;扬子鳄卵巢中TR的m RNA表达水平在繁殖期最高,在繁殖前期和繁殖后期相对较低;免疫组织化学染色显示,TR阳性反应主要位于卵泡的颗粒层细胞、卵泡膜、卵母细胞质和卵母细胞核中,其中颗粒层细胞阳性反应最明显。结论 TR基因的进化时间较早,因此序列保守性较高;TRα和TRβ对扬子鳄卵泡发育成熟具有重要作用;颗粒层细胞是TRs的功能作用主要位点。     

硒对AOM所致结肠癌模型大鼠肾上腺皮质束状带细胞组织化学的影响

目的观察硒对致癌剂氧化偶氮甲烷(azoxymethane,AOM)所致结肠癌模型大鼠肾上腺皮质束状带细胞组织化学的变化。方法随机将20只3周龄SPF级断乳雄性Sprague Dawley(SD)大鼠随机分4组:正常对照组、实验对照组、致癌剂前补硒组和致癌剂后补硒组。用AOM(15mg/kg)每周腹腔注射,连续2周,诱导大鼠结肠癌的形成。亚硒酸钠(Na2SeO3)水溶液(4mg/L)分别在AOM前、后干预,并持续至实验结束。各组均于34周取大鼠肾上腺,用组织化学的方法,观察大鼠肾上腺皮质束状带细胞组织化学的变化,对脂类、琥珀酸脱氢酶(SDH)和3β-羟类固醇脱氢酶(3β-HSD)进行染色,并图像分析。结果亚甲基蓝染色光镜下观察,可见AOM腹腔注射的大鼠结肠黏膜出现异常隐窝(aberrant crypt,AC)和异常隐窝灶(aberrant crypt foci,ACF)。组织化学显示,与正常对照组相比,实验对照组大鼠肾上腺皮质束状带细胞的SDH和3β-HSD的MOD值明显增加(P<0.05),脂类的MOD值明显减少(P<0.05);硒干预的各组与实验对照组相比,大鼠肾上腺皮质束状带细胞的SDH和3β-HSD的M...     

SHIV感染恒河猴十二指肠黏膜活检组织病理学观察

目的使用消化内窥镜对SHIV感染恒河猴十二指肠黏膜进行观察。方法SHIV感染恒河猴,消化内镜检查并进行活检取材,对活检标本进行固定,石蜡包埋,常规HE染色镜检。结果内镜大体观察胃黏膜和十二指肠黏膜基本上完整平滑,1例胃黏膜有弥漫性白斑,3例有小面积的出血;光镜下十二指肠黏膜完整未见脱落,固有层中有散在分布的慢性炎细胞,活检取材最深可达黏膜下层,未见其他特异性病理改变。结论不同毒株,不同感染时间的SHIV感染恒河猴和对照恒河猴的十二指肠黏膜活检组织HE染色组织结构清晰,呈慢性炎症改变,病理形态上没有明显的差异。     

小鼠早期胚胎发育期间TGF-β_1免疫组织化学定位

用兔抗人血小板TGF-β_1 N末端1—29氨基酸残基人工合成多肽抗血清作探针以及免疫荧光和免疫酶染色技术,分析了1—12天小鼠早期发育期间胚胎的TGF-β_1物质分布。结果表明,着床前胚胎包括卵裂细胞,桑椹胚和胚泡的ICM及滋养外胚层等细胞均显示TGF-β_1阳性免疫荧光染色。免疫酶染色还证明,沿囊胚腔顶部单层排列的原始内胚层细胞比邻近的ICM细胞有较深的染色反应。随着胚胎着床和进一步发育,7天龄胚胎中胚层早期形成阶段,紧靠中胚层一侧的外胚层胞质中含有浓集的棕色颗粒;各胚层的部分区域也存在着染色强度上差别。8—12天龄胚胎中,体节,心壁、间质细胞和肠道以及卵黄囊的脏壁中胚层均有显著的TGF-β_1免疫酶阳性物质。这些结果表明,着床前小鼠胚胎富含TGF-β_1物质,着床后的胚外组织,例如卵黄囊也为胚胎进一步发育提供了富含TGF-β_1物质的微环境;同时也提示,小鼠早期胚胎发育期间的胚泡形成,ICM细胞分化出原始内胚层,卵柱期中胚层形成,以及以后的神经管、体节和肢芽形成阶段等一系列形态发生和器官形成过程中,TGF-β_1可能是参与重要作用的一种生长调节因子。     

猴艾滋病急性期肠粘膜相关淋巴组织NK细胞表型及功能

目的研究猴艾滋病毒感染急性期恒河猴肠道相关淋巴组织（mucosal associated lymphoid tissues,MALTs）NK细胞亚群和功能变化。方法 SIV静脉感染恒河猴后,定期进行动物感染指标测定,并在感染后不同时间点取肠组织,分离派氏淋巴结单个核细胞（peyer＇s patch mononuclear cells,PPMC）和粘膜固有层单个核细胞（lamina propria mononuclear cells,LPMC）,进行T细胞和NK细胞表面抗体染色,流式分析。结果 SIV感染急性期MALTs CD56CD16＋NK细胞亚群比例增幅明显,同时细胞毒性功能增强;CD56-CD16-NK细胞亚群数量减少,功能无明显变化;CD56＋CD16＋和CD56＋CD16-NK细胞数量比例略有增加趋势,但免疫调节功能显著降低。结论SIV感染急性期恒河猴肠道MALTs中NK细胞脱颗粒作用增强,表型功能呈现出较强可塑性。该研究对探索艾滋病粘膜免疫机理、抗病毒治疗及药物研发具有参考意义。     

Immunocytochemical localization of annexin 5, a calcium-dependent phospholipid-binding protein, in rat endocrine organs

Annexin 5, a unique calcium- and phospholipid-binding protein, has been investigated for its specific distribution in rat endocrine organs by immunocytochemistry with a specific antiserum to recombinant rat annexin 5. Follicular epithelial cells and parafollicular cells of the thyroid gland, adrenocortical cells of the zona fasciculata and zona reticularis, luteal cells, testicular interstitial cells, and Sertoli cells were shown to contain annexin 5. To examine whether the synthesis of annexin 5 would be affected by a change in humoral signal, the distribution of annexin 5 in the anterior pituitary was examined three weeks after ovariectomy. The withdrawal of ovarian hormones induced huge castration cells in the anterior pituitary gland, which contained abundant annexin 5. Annexin 5 was not detected in the pineal gland, the parathyroid gland, the islet of Langerhans, the adrenal medulla, zona glomerulosa cells, and granulosa cells. Since annexin 5 was shown to exist in many of the endocrine tissues examined, to be localized in specific cell types, and to be abundant in castration cells, it is suggested that annexin 5 contributes to secretory cell functions, which may be common to endocrine cells secreting chemically different hormones.     

Morphology and ultrastructure of the adrenal gland in Bactrian camels (Camelus bactrianus)

In the present study, we examined the morphological features of the adrenal gland in Bactrian camel by means of digital anatomy, light and electron microscopy. Our findings testified that the gland was divided into three parts, capsule, cortex and medulla from outside to inside as other mammals, and the cortex itself was further distinguished into four zones: zona glomerulosa, zona intermedia, zona fasciculate and zona reticularis. Notably, the zona intermedia could be seen clearly in the glands from females and castrated males, whereas it was not morphologically clear in male. There was a great deal of lipid droplets in the zona fasciculate, while it was fewer in the zona glomerulosa and zona reticularis. The cytoplasm of adrenocortical cell contained rich mitochondria and endoplasmic reticulum. The adrenal medulla was well-developed with two separations of external and internal zones. The most obvious histological property of adrenal medulla cells were that they contained a huge number of electron-dense granules enveloped by the membrane, and so medulla cells could be divided into norepinephrine cells and epinephrine cells. Moreover, the cortical cuffs were frequently present in adrenal gland. Results of this study provides a theoretical basis necessary for ongoing investigations on Bactrian camels and their good adaptability in arid and semi-arid circumstances.     

Proliferation and distribution of adrenocortical cells in the gland of ACTH- or dexamethasone-treated rats

The effects of prolonged (7-day) ACTH and dexamethasone administrations on rat adrenocortical-cell turnover have been investigated by combined stereological and metaphase-arrest techniques. ACTH was found to increase the number of parenchymal cells in each adrenal zone; however, ACTH altered the cell distribution in the cortex, lowering their percentage in the zona glomerulosa (ZG) and zona fasciculata (ZF) and enhancing it in the zona reticularis (ZR). The cell birth-rate was markedly raised by ACTH exclusively in ZG and ZF. Dexamethasone notably decreased the number of ZF and ZR cells, without altering that of ZG cells. Moreover, dexamethasone increased the percentage of parenchymal cells in ZG and ZF, and lowered it in ZR. In the adrenal cortices of dexamethasone-administered animals, metaphases were virtually absent. These data indicate that ACTH increases the cell birthrate in ZG and possibly ZF, and enhances the centripetal migration of newly-formed cells and their accumulation in ZR. Dexamethasone inhibits both proliferation of adrenocortical cells in the outer cortical layers and their centripetal migration into ZR. Moreover, it appears to cause parenchymal-cell loss in the inner adrenocortical layers.     

Cytochrome P-45011 beta and P-450scc in adrenal cortex: zonal distribution and intramitochondrial localization by the horseradish peroxidase-labeled antibody method

In an attempt to elucidate the regulation mechanism(s) of adrenocortical steroidogenesis, cytochrome P-450scc and cytochrome P-45011 beta were localized in bovine adrenal glands by the direct peroxidase-labeled antibody method. At the light microscopic level, parenchymal cells of the zona fasciculata and the zona reticularis stained heavily for both cytochromes, while the parenchymal cells of zona glomerulosa stained lightly for both. At the electron microscopic level, these two cytochromes were associated with the matrix side of the inner mitochondrial membranes of parenchymal cells from all three zones of the adrenal cortex. The association of cytochrome P-450 with the inner mitochondrial membrane, in a manner similar to that previously reported for adrenodoxin and adrenodoxin reductase (F Mitani, Y Ishimura, S Izumi, K Watanabe, Acta Endocrinol 90:317, 1979), establishes that the steroid monooxygenase systems exist at this site. The degree of immunocytochemical staining within a single cell varied from one mitochondrion to another: some stained intensely along the entire inner membrane, including the cristae, some stained only along segments of the inner membrane, and some did not stain at all. This heterogeneity in staining was observed in mitochondria stained in situ as well as in isolated mitochondria. These findings suggest that there is a heterogeneity in steroidogenesis among mitochondria contained within a single cell of the adrenal cortex.     

Sex differences in adrenocortical structure and function

Summary Adrenal glands of adult male hamsters are larger and secrete more cortisol than those of females. Stereology was therefore used to study zonal and cellular aspects of development of the adrenal cortex of male and female hamsters. Adrenal glands were studied at weekly intervals from day 21 to day 77 of postnatal ontogenesis. Within this period, body weight did not differ significantly between the sexes. During development, absolute and relative adrenal weights were higher in males; their zona glomerulosa (ZG), zona fasciculata (ZF) and zona reticularis (ZR) become markedly larger than those in females. No marked changes in the volume of individual ZG cells occurred although ZF cells and ZR cells become larger in male than female animals. The total number of adrenocortical cells increased within the period studied, a greater increase being observed in ZG and ZF in males. No distinct sex difference was observed in the number of ZR cells throughout development. From day 56 of postnatal life the adrenal cortex of male hamster contained more parenchymal cells than the female gland. These results thus indicate that sex differences in hamster adrenal cortex depend upon changes in number and size of parenchymal cells.Supported in part by a grant from the Committee of Zoology, Polish Academy of Sciences     

Immunocytochemical localization of adrenodoxin in bovine adrenal cortex by protein A-gold technique

To investigate the intracellular localization of the enzymes that are involved in steroid hormone synthesis, an immunocytochemical study of the distribution of adrenodoxin in cells of the bovine adrenal cortex was carried out by the post-embedding immunostaining method and by immuno-cryoultramicrotomy in combination with the protein A-gold technique. Gold particles were seen on the matrix and the inner membrane of all the mitochondria examined, which have typical vesicular or tubulo-vesicular cristae, in parenchymal cells of the zona fasciculata and the zona reticularis. Gold particles were distributed homogeneously among the mitochondria. The density of gold particles on mitochondria in the parenchymal cells of the zona glomerulosa was lower than that of the zona fasciculata, which was similar to that of the zona reticularis. Gold particles were also seen on round, electron-dense intramitochondrial bodies in the parenchymal cells. The intramitochondrial bodies were abundant in the zona glomerulosa and the outer region of the zona fasciculata.     

Immunocytochemical localization of adrenodoxin in bovine adrenal cortex by protein A-gold technique

Summary To investigate the intracellular localization of the enzymes that are involved in steroid hormone synthesis, an immunocytochemical study of the distribution of adrenodoxin in cells of the bovine adrenal cortex was carried out by the post-embedding immunostaining method and by immuno-cryoultramicrotomy in combination with the protein A-gold technique. Gold particles were seen on the matrix and the inner membrane of all the mitochondria examined, which have typical vesicular or tubulo-vesicular cristae, in parenchymal cells of the zona fasciculata and the zona reticularis. Gold particles were distributed homogeneously among the mitochondria. The density of gold particles on mitochondria in the parenchymal cells of the zona glomerulosa was lower than that of the zona fasciculata, which was similar to that of the zona reticularis. Gold particles were also seen on round, electron-dense intramitochondrial bodies in the parenchymal cells. The intramitochondrial bodies were abundant in the zona glomerulosa and the outer region of the zona fasciculata.Supported by a Grant-in-Aid for Scientific Research on Priority Areas (no. 63635505) from the Ministry of Education, Science and Culture of Japan     

Class II MHC-expressing cells in the rat adrenal gland defined by monoclonal antibodies

 The detailed distribution and heterogeneity of various immunocompetent cells were characterized in the normal adrenal gland of the rat, with special emphasis on major histocompatibility complex (MHC) class II-expressing cells and macrophages. All adrenals contained at least two different populations of cells reactive with the dendritic cell or the macrophage antibodies. These cells were clearly distinguished from adrenal parenchymal cells by their morphology and location. The majority of dendritic cells were immunoreactive for the MHC class II (Ia) antigen (MRC OX6) and/or the dendritic cell antibodies (MRC OX62), and negative for the macrophage antibodies (ED1, ED2, and/or MRC OX42), whereas the main population of macrophages was immunonegative for the former antibodies and positive for the latter. The OX62-positive cells and the OX42-labeled cells occurred exclusively throughout the medulla. The cellular density of dendritic cells in the adrenal cortex was significantly higher than that of macrophages. Double-immunoperoxidase staining for ED1 and OX6 revealed that positively stained cells could be classified into the following categories: ED1⁺OX6⁺, ED1⁺OX6⁻, and ED1⁻OX6⁺. More then 40% of OX6⁺ cells were immunoreactive for ED1 in the zona glomerulosa, while approximately 15%, 20%, and 30% of OX6⁺ cells were positive for ED1 in the zona fasciculata, zona reticularis and medulla, respectively. ED1⁺ED2⁻ cells were more frequently detected in the zona glomerulosa than in other adrenal zones. Only a few ED1⁻ED2⁺ cells were located in the zona glomerulosa, whereas a large number of them were found in the zona fasciculata. In the zona reticularis and medulla, ED1⁺ED2⁺, ED1⁺ED2⁻, and ED1⁻ED2⁺ cells were detected in the ratio 2:1:3. Our rsults suggest that dendritic cells and macrophages mature during their migration within the adrenal gland. These immunocompetent cells may contribute to a paracrine regulation of adrenal function under physiological conditions. Accepted: 3 November 1997     

Fine structure of the rabbit adrenal cortex and the effects of short-term ACTH administration

Summary The fine structure of the rabbit adrenal cortex was investigated. The parenchymal cells display the ultrastructural features of steroid-producing cells, and also contain numerous electron-dense bodies frequently located near intercellular canaliculi, which open into the subendothelial space. Short-term ACTH-administration induced a noticeable decrease in the volume of the lipid compartment in the cells of all three cortical zones and a significant increase in the volume of dense bodies in the cells of zona fasciculata and zona reticularis. The hypothesis that these dense bodies are secretory granules is discussed in the light of biochemical evidence showing that ACTH increases the concentration of both corticosterone and cortisol in the decapsulated-enucleated adrenal homogenate and does not affect the activity of two lysosome-marker enzymes (i.e., acid phosphatase and -glucuronidase).     

Effects of unilateral adrenalectomy on the remaining adrenal cortex of adrenocorticotropic hormone-treated male and female hamsters

Sham-operated and unilaterally adrenalectomized male and female hamsters were administered 25 micrograms adrenocorticotropic hormone (ACTH) for 5 days after the operation in order to examine the effects of ACTH on compensatory adrenal growth. In ACTH-treated male and female hamsters, unilateral adrenalectomy did not change the relative weight of the remaining adrenal. There were no significant differences in the volumes of the adrenocortical zones and their parenchymal cells, as well as in the number of adrenocortical cells per gland if compared with unilaterally adrenalectomized and sham-operated ACTH-treated male hamsters, while 3H-thymidine incorporation per gland was lower in monoadrenalectomized animals. On the contrary, in ACTH-treated females, unilateral adrenalectomy resulted in a significant hypertrophy of zona fasciculata cells and in an enhanced 3H-thymidine uptake by the remaining gland. These findings stress the existence of notable sex-related differences in the compensatory adrenal growth in hamsters.