大鼠颌下腺促性腺激素释放激素及其mRNA的免疫组织化学与原位杂交 …

用免疫组织化学及原位杂交法,研究了促性腺激素释放激素及其ｍＲＮＡ在大鼠颌下腺的分布。结果显示,大鼠颌下腺的浆液腺泡的上皮细胞,各级导和的上皮细胞及副交感神经节细胞均呈促性腺激素释和激素免疫反应阳性,阳生反应物质分布在胞质,胞核呈阴性反应。     

大鼠颌下腺及其离体培养细胞脱氢表雄酮（DHEA）的免疫组织化学定位

用免疫组织化学ABC法,研究了颌下腺及无血清培养的颌下腺上皮细胞DHEA的定位,结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞及各级导管上皮细胞均呈DHEA免疫反应阳性,无血清培养腺上皮细胞也呈DHEA免疫反应阳性,阳性物质分布于胞质,胞核呈阴性反应,此结果提示：大鼠颌下腺能自身合成DHEA,DHEA对消化功能可能具有重要的调节作用。     

胍基丁胺在成年大鼠组织中的表达和分布

目的观察胍基丁胺（agmatine,AGM）在成年Wistar大鼠全身组织的表达和分布。方法采用免疫组织化学法,用自制的抗胍基丁胺多克隆抗体检测胍基丁胺在成年Wistar大鼠全身组织中的表达分布情况。结果胍基丁胺在胃、十二指肠、空肠、回肠、结肠的粘膜腺上皮细胞胞质内呈强阳性表达;胍基丁胺在大脑皮层、丘脑、海马、延髓的神经元胞质和神经轴突内呈强阳性表达;胍基丁胺在子宫、输卵管、前列腺的腺上皮细胞和睾丸的生精细胞胞质内呈强阳性表达;胍基丁胺在肝细胞、肾上腺皮髓质细胞胞质内呈阳性表达,在肾脏、脾脏呈弱阳性表达;胍基丁胺在肺、阴茎的微血管平滑肌细胞胞质内呈强阳性表达,但在心脏、骨骼肌、食道、主动脉的肌细胞内未见其表达。结论胍基丁胺在成年Wistar大鼠体内广泛分布,可能具有重要的生理作用。     

大鼠颌下腺促性腺激素释放激素及其mRNA的免疫组织化学与原位杂交研究

用免疫组织化学及原位杂交法,研究了促性腺激素释放激素及其ｍＲＮＡ在大鼠颌下腺的分布。结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞,各级导管的上皮细胞及副交感神经节细胞均呈促性腺激素释放激素免疫反应阳性,阳性反应物质分布在胞质,胞核呈阴性反应。颌下腺的浆液性腺泡上皮细胞,各级导管上皮细胞同样被检测到很强的促性腺激素释放激素ｍＲＮＡ杂交信号。以上结果提示,大鼠颌下腺能自身合成促性腺激素释放激素,促性腺激素释放激素对消化功能可能有重要调节作用。     

半胱胺促进空怀奶山羊乳腺发育的试验研究

目的：研究半胱胺对空怀奶山羊乳腺发育的影响。方法：选6只空怀奶山羊进行自身前后对照试验。对照期饲喂基础日粮,试验期饲喂基础日粮加CS。结果：与对照期相比,试验期山羊乳腺组织的DNA含量明显升高,RNA含量明显下降,故RNA／DNA值显著下降。试验期血浆GH和PRL水平分别提高61．6％和31．6％。组织学观察发现,对照期乳腺的腺泡很少发育,几乎看不到乳导管;试验期乳腺有明显导管生长和少量腺泡发育。结论：半胱胺能够在一定程度上促进空怀奶山羊乳腺发育。     

黑缘烟蟋螽丝腺结构

【目的】蟋螽是直翅目中唯一具有吐丝筑巢行为的类群。本研究旨在探讨蟋螽丝腺的结构特点。【方法】应用解剖学观察、免疫荧光、苏木精-伊红染色、PAS苏木精染色、扫描电镜和透射电镜等方法从细胞水平对黑缘烟蟋螽Capnogryllacris nigromarginata丝腺的显微与超微结构进行了观察。【结果】黑缘烟蟋螽丝腺由导管和腺泡构成。腺泡由鞘细胞延伸形成的结缔组织鞘包围。腺泡的主体有4种细胞,分别为Ⅰ型分泌细胞、Ⅱ型分泌细胞、围细胞和腔细胞。Ⅰ型和Ⅱ型分泌细胞为大的腺细胞,形状不规则。分泌细胞细胞核很大,胞质内有大量的内质网和分泌颗粒。Ⅰ型分泌细胞靠近腺泡中心,PAS-苏木精染色表明Ⅰ型分泌细胞内含糖蛋白,Ⅱ型分泌细胞在腺泡外周,位于Ⅰ型分泌细胞与围细胞或结缔组织鞘之间。腔细胞分散在分泌细胞之间,包围形成胞外运输分泌物的通道。围细胞与鞘细胞接触,具有由细胞膜内陷形成的微绒毛腔,胞质内有大量的线粒体。围细胞微绒毛腔与腔细胞包围的细胞外运输通道相连,分泌细胞分泌的颗粒聚集在分泌细胞和胞外运输通道之间的连接处,并将分泌物排出至胞外运输通道。多个腺泡的胞外运输通道汇集到由单层细胞组成的丝腺导管。单层导管细胞靠近管腔外围具有规则排列的质膜内陷和大量伸长的线粒体;靠近管腔的一侧具连续的细胞膜突起,在导管壁的表皮下紧密排列。【结论】黑缘烟蟋螽丝腺分泌细胞分为Ⅰ型分泌细胞和Ⅱ型分泌细胞。分泌物质产生及分泌过程依次经过分泌细胞、腔细胞包围的胞外通道、分支导管、总导管和唾窦。其中在腺泡细胞之间,分泌物向外运输过程中,围细胞微绒毛腔的微丝束可能对分泌物的外排提供推动力。     

不同年龄与不同时期林麝香腺组织结构观察

本研究旨在探究林麝(Moschus berezovskii)在不同年龄和泌香期与非泌香期香腺的组织结构变化,为深入研究林麝香腺发育和麝香分泌机制提供基础资料。收集6只雄性林麝的香腺组织,包括泌香期2岁龄林麝1例,非泌香期6月龄、2岁龄、4岁龄、6岁龄和8岁龄林麝各1例,采用大体解剖、石蜡切片及常规H.E染色技术,对香腺的形态、组织特征及腺泡直径进行了比较分析。结果显示,林麝香腺位于腹部阴囊与腹脐之间,与阴囊的距离约为4.5 cm。根据功能,香腺可划分为香囊部和腺体部。负责分泌麝香液的腺体组织为肉眼可见的白色颗粒,嵌入腺体部肌层深处并环绕香囊颈分布。6月龄林麝香腺已具有成熟腺体结构,但腺上皮仍处于休止状态;成年林麝香腺的腺泡则增大且数量增多。成年林麝泌香期腺泡被挤压成团状分布,上皮游离面破碎不整,腺泡腔内含有大量明显的颗粒状分泌物与细胞碎屑混合堆积;非泌香期腺泡直径显著大于泌香期(P < 0.01),2岁林麝腺泡表现为紧密排列的椭球形,而4岁及以上的腺泡则呈不规则团状分布,腔内可见颗粒物,且4岁、6岁和8岁的腺泡直径无显著差异(P > 0.05)。林麝香腺组织结构的变化反映了泌香能力与生长阶段的关系。     

红腹锦鸡胃肠道内分泌细胞的免疫组织化学定位

应用ABC免疫组织化学方法,对红腹锦鸡(Chrysolophus pictus)胃肠道5-羟色胺(5-hydroxtryptamine,5-HT)、胃泌素(gastfin,GAS)、生长抑素(somatostatin,ss)3种内分泌细胞的分布密度和形态进行了观察.结果显示,5-HT细胞在空肠和直肠分布密度最高,回肠和盲肠次之,十二指肠较少,腺胃和肌胃最少;GAS细胞在十二指肠和直肠分布密度最高,其次是空肠和盲肠,腺胃部最低,肌胃则呈免疫阴性;SS细胞数量较少,在直肠、盲肠处分布密度相对高,其次是十二指肠和空肠,腺胃部最低,肌胃则呈免疫阴性.3种内分泌细胞的形态多样,有圆形、椭圆形、锥体形、杆状和不规则形,其中以圆形、椭圆形为主.细胞分布于固有膜、黏膜上皮细胞基部、黏膜上皮细胞之间、腺泡上皮细胞基部或腺泡上皮细胞之间.红腹锦鸡胃肠道内分泌细胞的形态与其内、外分泌功能是相适应的.     

人和大鼠胃窦部神经内分泌细胞分布和形态学的比较研究

目的探讨人和大鼠胃窦部神经内分泌细胞的分布和形态学特征。方法采用免疫细胞化学方法,检测人和大鼠胃窦部粘膜内生长抑素细胞(D细胞)、胃泌素细胞(G细胞)、5-羟色胺细胞(5-HT细胞)、嗜铬粒素A细胞(CgA细胞)的分布。结果人和大鼠的G、D细胞的特征是都具有细胞突起,但是在细胞密度及其分布上是不同的;5-HT细胞的分布在两组稍有不同,在大鼠胃窦部,大多数5-HT细胞位于腺体基部,而人的5-HT细胞主要在间质,少数位于腺上皮内;在两组中,CgA细胞几乎布满整个胃粘膜,其数量也高于其他神经内分泌细胞,CgA细胞形态多样,胞质内充满细小颗粒。结论1)人与大鼠的G、D细胞通常都伴有突起,但其分布不同。2)5-HT细胞形态多样,分布于间质和腺上皮内。3)CgA细胞特征是胞质内充满细小颗粒,细胞形态多样,几乎布满整个粘膜。     

消乳散结胶囊治疗兔乳腺增生的实验研究

目的:观察消乳散结胶囊(XRSJ)在治疗兔乳腺增生中的作用,探讨其治病机理。方法:将90只日本大耳兔随机分为6组,分别为XRSJ高、中、低剂量组,逍遥丸组,模型和正常对照组,每组15只,前5组用肌内注射己烯雌酚、黄体酮1个月建立兔乳腺增生模型。造模完成后,行XRSJ灌胃给药,分别取XRSJ给药前、XRSJ给药3个月后及XRSJ停药3个月后作为3个时间观测点。观察乳房高度变化;乳腺光、电镜形态学变化;放射免疫法监测血清雌激素水平。结果:与治疗前及模型对照组相比,中、高剂量XRSJ治疗能明显降低乳房高度(P<0.05);降低增生乳腺小叶腺泡数目、导管腺上皮细胞层数、结缔组织和毛细血管数量(P<0.05);减少乳腺上皮细胞胞质内线立体、高尔基体、粗面内质网数量(P<0.05),使部分增生细胞调亡;显著降低血清雌二醇(E2)含量(P<0.01),停药3个月后治疗效果稳定无反弹。结论:XRSJ治疗具有减轻乳腺组织增生病变、降低E2水平、对兔乳腺增生具有显著的治疗作用。     

Use of peanut lectin and rat mammary stem cell lines to identify a cellular differentiation pathway for the alveolar cell in the rat mammary gland.

The presence of the carbohydrate receptor for PNL has been used to identify the previously described morphological types of epithelial cell produced as the stem cell line rat mammary 25 (Rama 25) differentiates to casein secretory alveolar-like cells in vitro. Thus when cultures of the epithelial stem cell line Rama 25 are treated with neuraminidase, fluorescently-conjugated PNL fails to stain cuboidal cells, stains weakly grey cells, and stains strongly the surface of dark cells. When superconfluent cultures of Rama 25 are treated with dimethyl sulfoxide or retinoic acid and prolactin, estradiol, hydrocortisone, and insulin to induce differentiation to alveolar cells, PNL stains strongly the untreated surfaces of droplet cells and casein-secreting vacuolated cells. PNL-staining of the derivative cell lines with truncated cellular pathways, and quantitative binding of [125I]-labeled PNL to the cultured cells are consistent with this cellular staining pattern. The presence of the carbohydrate receptor for peanut lectin (PNL) has also been used to identify specific epithelial cell types in different mammary structures of the developing rat mammary gland, as they differentiate to casein secretory alveolar cells in vivo. Thus when different structures of the developing rat mammary gland are treated with neuraminidase, peroxidase-conjugated PNL fails to stain histochemically the majority of epithelial cells in ducts, stains the cytoplasm of the majority of epithelial cells in terminal end-buds (TEBs), and stains strongly the luminal surfaces of the majority of epithelial cells in alveolar buds (ABs). PNL also stains the untreated luminal surfaces of alveolar cells, whether or not the cells can be stained with a monoclonal antibody to rat beta-casein. Stimulation of mammary differentiation by an analogue of ethyl retinoate or by perphenazine causes cells in end-buds to bind PNL without the necessity for their desialylation similar to that seen in casein secretory alveoli of lactating rats. In conclusion the different interconverting cell types of Rama 25 which form a pathway to casein-secretory cells in vitro are thus equated with recognisable epithelial cell types in vivo. These results suggest that casein-secretory cells in vivo are generated by similar successive interconversions between the major epithelial cell types present in the different mammary structures in the order: ducts, TEBs, ABs, alveoli, and secretory alveoli.     

Expression and localisation of GLUT1 and GLUT12 glucose transporters in the pregnant and lactating rat mammary gland

Glucose plays a major role in mammary gland function during lactation as it is used both as a fuel and as a precursor of milk components. In rats, previous studies have shown that the facilitative glucose transporter GLUT1 is expressed in mammary epithelial cells. We have used confocal immunofluorescence to localise GLUT1 and GLUT12, a recently identified member of the sugar transporter family, in pregnant and lactating rat mammary gland. GLUT12 staining was observed in the cytoplasm of mammary epithelial cells at day 20 of pregnancy, and at 1 and 6 days postpartum. Furthermore, GLUT12 staining was present at the apical plasma membrane of epithelial cells during lactation. In contrast, GLUT1 protein localised to the cytoplasm and basolateral surface of mammary epithelial cells. Forced weaning resulted in decreased cytoplasmic GLUT1 staining intensity, but no change in GLUT12 staining. The results suggest a possible role for GLUT12 in the metabolism of mammary epithelial cells during pregnancy and lactation.     

Expression, activity, and localization of hormone-sensitive lipase in rat mammary gland during pregnancy and lactation

We examined the presence of hormone-sensitive lipase (HSL) in mammary glands of virgin, pregnant (12, 20, and 21 days), and lactating (1 and 4 days postpartum) rats. Immunohistochemistry with antibody against rat HSL revealed positive HSL in the cytoplasm of both alveolar epithelial cells and adipocytes. In virgin rats, immunoreactive HSL was observed in mammary adipocytes, whereas diffuse staining was found in the epithelial cells. Positive staining for HSL was seen in the two types of cells in pregnant and lactating rats. However, as pregnancy advanced, the staining intensity of immunoreactive HSL increased in the epithelial cells parallel to their proliferation, attaining the maximum during lactation. An immunoreactive protein of 84 kDa and a HSL mRNA of 3.3. kb were found in the rat mammary gland as in white adipose tissue. Both HSL protein and activity were lower in mammary glands from 20 and 21 day pregnant rats than from those of virgin rats, although they returned to virgin values on days 1 and 4 of lactation. Mammary gland HSL activity correlated negatively to plasma insulin levels. Immunoreactive HSL and HSL activity were found in lactating rats' milk. The observed changes indicate an active role of HSL in mammary gland lipid metabolism.     

The 24p3 gene is induced during involution of the mammary gland and induces apoptosis of mammary epithelial cells

To understand the molecular mechanism of mammary gland involution we identified involution-induced clones by differential screening of a mouse mammary gland cDNA library. Characterization of clones by sequencing and Northern analysis showed that expression of 24p3 was induced during involution of the mammary gland. RNA in situ hybridization showed that it was mainly expressed in the secretory epithelial cells surrounding the lumen of the mammary gland alveoli. Induction of 24p3 was also observed in apoptotic HC11 mammary epithelial cells under serum starvation. In these cells, dexamethasone increased 24p3 gene expression four-fold. Transient expression of 24p3 increased the percentage of apoptotic cells 3- to 4-fold over a period of 3 days after transfection. This study provides evidence that overexpression of 24p3 gene can induce apoptosis of mammary epithelial cells.     

大鼠自发性乳腺癌瘤株的建立和病理学研究

[目的]试图建立大鼠自发性乳腺癌产生C型肿瘤病毒的瘤株,探讨大鼠自发性乳腺癌病理学形态特点与C型肿瘤病毒形态学结构。[方法]将大鼠乳腺原发性癌用细胞悬液法和瘤块法交替传代建立瘤株;大鼠乳腺原发性癌和移植性癌分别用10%福尔马林液固定,石蜡包埋切片,HE和Ag染色,光镜观察;分别取大鼠乳腺原发性癌和移植性癌用2.5%戊二醛与1%锇酸双重固定,Epon812包埋,超薄切片,醋酸双氧铀和柠檬酸铅双重染色,透射电镜观察。[结果]大鼠乳腺原发性癌细胞体积较小,直径为10μm,可见小球状结构,乳头状结构,弯曲的条索状与肾小球样结构形成典型的癌性腺管,Ag染色显示癌细胞由网状纤维分隔成不规则巢状,表明大鼠乳腺癌起源于上皮组织,其组织学类型为大鼠乳腺上皮性间变型腺癌。移植性癌细胞呈圆型、立方形、多边形,出现瘤巨细胞与苍白细胞等多种形态,排列成巢状、片状、小梁状、乳头状、小球状、癌性腺管和裂隙状结构,异形性显著。癌细胞形态不规则,细胞核大,核仁增多,胞质中有聚集成束的张力原纤维,罕见分泌颗粒,细胞间存在原始腺腔和桥粒;在瘤细胞间与胞浆内发现了C型肿瘤病毒颗粒,其形态学为球形,直径100nm,外部有囊膜、内膜、中央核心。[结论]首次建立了大鼠自发性乳腺癌产生C型肿瘤病毒的瘤株,其组织学类型为大鼠乳腺上皮性间变型腺癌,病理学形态是多态性的。     

Membranes of mammary gland : XV. 5-Thio- -glucose decreases lactose content and inhibits secretory vesicle maturation in lactating rat mammary gland

Short-term administration of the glucose analog 5-thio- -glucose to primiparous lactating rats reduced mammary tissue lactose concentrations to half of control levels. Treatment with colchicine alone caused slight reductions in mammary tissue lactose content. These treatments did not alter the morphology or degree of development of rough endoplasmic reticulum or Golgi apparatus, but did cause alterations in secretory vesicles. In mammary tissue from untreated lactating animals, large, swollen secretory vesicles were abundant in apical regions of epithelial cells. After thioglucose administration secretory vesicles in the apical cytoplasm were smaller and were more densely packed with contents. While administration of colchicine alone caused accumulation of large numbers of nearly fully swollen vesicles, treatment with both colchicine and thioglucose induced accumulation of smaller, less fully developed secretory vesicles which contained morphologically recognizable casein micelles. Mammary tissue from late gestation rats was low in lactose; vesicles in this tissue resembled secretory vesicles in tissue from rats treated with thioglucose in that they were small and densely packed. These observations suggest that lactose, an osmoregulator in mammary gland, is transferred from Golgi apparatus to the apical cell surface within secretory vesicles. Lactose appears to be important for secretory vesicle maturation in mammary epithelial cells.     

Expression of M-N#1, a histo-blood group B-like antigen, is strongly up-regulated in nonapoptosing mammary epithelial cells during rat mammary gland involution.

Antibodies against the histo-blood group B-like antigen M-N#1 efficiently block the growth in vivo of rat mammary carcinoma cells that bear the antigen (Sleeman et al., 1999, Oncogene 18, 4485--4494). To try to understand the function of the M-N#1 antigen, we investigated when and where the antigen is expressed during the normal function of the rat mammary gland. Expression was virtually only seen during mammary gland involution. Here, strong expression of the antigen was observed in mammary epithelial cells, beginning around 2 days postweaning and lasting throughout the involution process. Dexamethasone treatment of animals postlactation inhibited alveolar collapse and remodeling in the mammary gland but inhibited neither the apoptosis of mammary epithelial cells nor the expression of the M-N#1 antigen. We show that up-regulation of carbohydrate antigens is not a general phenomenon during mammary gland involution, and thus that M-N#1 antigen expression is specifically regulated. Up-regulation of alpha(1,2)fucosyltransferase A, an enzyme required for M-N#1 antigen synthesis, is at least partly responsible for regulated M-N#1 antigen expression postlactation. Most significantly, we observed that the M-N#1 antigen is virtually exclusively expressed on nonapoptosing epithelial cells in the involuting mammary gland. These data suggest that M-N#1 antigen expression might either provide a survival function and/or be expressed in epithelial cells that are destined to grow and remodel mammary duct structures.