Gene expression of connective tissue growth factor in adult mouse

The connective tissue growth factor (CTGF) is a well-known fibroblast mitogen and angiogenic factor that plays an important role in bone formation during embryogenesis. In the adult, CTGF is involved in wound healing as well as fibrotic and vascular disease. However, little is known about its physiological functions under non-pathological conditions in the adult organism. Here, we describe the cellular site of the CTGF mRNA expression in adult male and female mice as revealed by in situ hybridization histochemistry. Strong and persistent CTGF gene expression was particularly prominent in the mesenchyme of the cardiovascular system (aorta, auricular tissue, renal glomeruli), the mesenchyme surrounding the ovarian follicles or the testicular tubes in the gonadal tissue, and the subcapsular mesenchyme bordering densely innervated parts of whisker hair vibrissae. CTGF hybridization signals were not observed in the mesenchyme of many other organs including gut, muscle, liver or most parts of the lymphatic tissue. Strong expression was also present in the primary (early) ovarian follicles, the epithelium of the deep uterine glands and on myenteric ganglia neurons. These data suggest a selective and continuous mesenchymal function in the gonads and those tissues attracting very strong vascular supply or peripheral innervation. CTGF may also be involved in the cyclical proliferation of the uterine gland epithelium and in the early stages of follicular maturation, as well as in the neuropeptide regulation in the gut, cardiovascular and renal systems.     

Gene expression of connective tissue growth factor in adult mouse

The connective tissue growth factor (CTGF) is a well-known fibroblast mitogen and angiogenic factor that plays an important role in bone formation during embryogenesis. In the adult, CTGF is involved in wound healing as well as fibrotic and vascular disease. However, little is known about its physiological functions under non-pathological conditions in the adult organism. Here, we describe the cellular site of the CTGF mRNA expression in adult male and female mice as revealed by in situ hybridization histochemistry. Strong and persistent CTGF gene expression was particularly prominent in the mesenchyme of the cardiovascular system (aorta, auricular tissue, renal glomeruli), the mesenchyme surrounding the ovarian follicles or the testicular tubes in the gonadal tissue, and the subcapsular mesenchyme bordering densely innervated parts of whisker hair vibrissae. CTGF hybridization signals were not observed in the mesenchyme of many other organs including gut, muscle, liver or most parts of the lymphatic tissue. Strong expression was also present in the primary (early) ovarian follicles, the epithelium of the deep uterine glands and on myenteric ganglia neurons. These data suggest a selective and continuous mesenchymal function in the gonads and those tissues attracting very strong vascular supply or peripheral innervation. CTGF may also be involved in the cyclical proliferation of the uterine gland epithelium and in the early stages of follicular maturation, as well as in the neuropeptide regulation in the gut, cardiovascular and renal systems.     

周期性张应力下人牙周膜成纤维细胞结缔组织生长因子的表达

背景：前期研究发现,周期性张应力在一定时间内可以诱导人牙周膜成纤维细胞增殖。 目的：观察周期性张应力对人牙周膜成纤维细胞表达结缔组织生长因子的影响;明确JNK、p38MAPK、PI3K信号通路在周期性张应力诱导人牙周膜成纤维细胞表达结缔组织生长因子过程中的作用。 方法：采用多通道细胞牵张应力加载系统对体外培养的人牙周膜成纤维细胞分别给予周期性张应力刺激1,6,12,24 h,并以未加力组为对照组。对加力12 h的细胞分别添加JNK、p38MAPK、PI3K信号通路特异性抑制剂预处理,并与未加抑制剂的细胞作对比。应用ELISA法检测培养细胞分泌到上清液中的结缔组织生长因子蛋白;应用荧光定量PCR技术检测细胞结缔组织生长因子mRNA的表达。 结果与结论：加载周期性张应力组与对照组相比较,1 h人牙周膜成纤维细胞表达结缔组织生长因子开始增强、6 h表达明显增强,12 h达最高峰值、24 h表达开始下降。加入JNK信号通路特异性抑制剂后,人牙周膜成纤维细胞表达结缔组织生长因子出现下降;而加入p38MAPK信号通路和PI3K信号通路的特异性抑制剂后结缔组织生长因子表达未发生明显改变。提示在一定时间范围内,周期性张应力引起结缔组织生长因子mRNA与蛋白水平的表达与时间呈依赖性升高;其后随着时间的延长,结缔组织生长因子的表达则开始下降。周期性张应力通过JNK通路的介导调控人牙周膜成纤维细胞结缔组织生长因子的表达。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Distribution of macromolecular components in calf dermal connective tissue

Distribution of type I and type III collagens, glycosaminoglycans and non-collagenous glycoprotein(s) in calf skin has been investigated in five horizontally sliced layers of 250 microns thickness from the surface to the bottom. The upper dermis (the top three layers) consisting of fine collagen fibers had a higher ratio of hyaluronic acid/dermatan sulfate than the lower layers where coarse collagen fibers are mainly located. The ratios of total glycosaminoglycans/collagen and glycoprotein(s) collagen were higher in the upper dermis, while dermatan sulfate/collagen remained constant throughout the dermis. Relative content of type III collagen/total collagen (type I + III collagens) was also higher in the upper dermis. Possible involvement of these macromolecular constituents in collagen fibrillogenesis in vivo is also discussed.     

Resorption of connective tissue in the gingiva of the mouse incisor

During eruption of the mouse incisor part of the periodontal ligament moves along with the tooth in occlusal direction and is degraded in the gingiva, just apically of the junctional epithelium. When studied with the electron microscope some connective tissue cells (fibroblasts) showed disorganization of both nucleus and cytoplasm. In other cells vacuoles were observed containing partly degraded cellular constituents. Sometimes within these vacuoles electron dense material with the appearance of clumped chromatin was observed. On the basis of this observation it is concluded that heterophagocytosis contributes to the process of fibroblast breakdown. The ultrastructure of these heterophagic cells resembled that of fibroblasts. Collagenous fibrils were observed within cytoplasmic vacuoles of fibroblasts indicating that collagen is phagocytosed and probably digested in the lysosomal apparatus. Part of the intercellularspace was filled with a homogeneous material of moderate electron density, intermingled with some collagenous fibrils. Within some cells of the junctional epithelium partly degraded material was observed which may indicate that the epithelium contributes to the removal of residual products of connective tissue degradation.     

褪黑素减轻哮喘小鼠结缔组织生长因子的表达和抑制气道重构

目的探讨褪黑素(MT)对哮喘小鼠结缔组织生长因子(CTGF)的影响及其在气道重构中的作用。方法将小鼠随机分成对照组、哮喘模型组、褪黑素治疗组(MT组)、地塞米松治疗组(DXM组),每组10只。用卵蛋白(OVA)致敏小鼠并反复雾化吸入OVA2周和4周,MT组和DXM组小鼠分别给予腹腔注射MT、DXM。HE染色观察肺组织气道中嗜酸性粒细胞;Masson三色染色观察气道周围胶原沉积;PAS染色观察气道黏液分泌;测定单位气道面积基底膜周径(Pbm)、管壁总面积(WAt)、内壁面积(WAi)、平滑肌面积(WAm)、胶原面积(Wcol)、黏液面积;用免疫组织化学检测肺组织中CTGF表达水平。结果哮喘模型组WAt/Pbm、WAi/Pbm、WAm/Pbm、Wcol/Pbm、黏液分泌面积及CTGF表达显著高于对照组,给予MT、DXM可显著缓解上述指标的升高。结论MT能降低哮喘小鼠肺组织CTGF表达水平,部分抑制气道重构,其调节作用与地塞米松相当。     

Alterations of the connective tissue components induced by β-aminopropionitrile

The ultrastructural and biochemical alterations induced by β-aminopropionitrile on aorta, lung, and skin of 7-day-old chicks have been studied. The inhibition of elastin formation by β-aminopropionitrile was associated with: (i) apposition of elastin on the old fiber in the form of button-like appendices; (ii) absence of microfibrils around this abnormally deposited elastin; (iii) presence of ruthenium red and toluidine blue O-positive material within the lathyritic elastin; (iv) increase of proteoglycan content; (v) increase of the mean diameter of collagen fibers; (vi) increased vesiculation of the endoplasmic reticulum of both fibroblasts and smooth muscle cells in aorta. The role of lysyl oxidase in the assembly of elastin and collagen fibers is discussed. Particular attention has been paid to the relationship between elastin, microfibrils, and proteoglycans in the formation of the elastic fiber.     

Effect of formalin fixation on the ultrastructure of connective tissue components

Electron microscopic examinations of fibrillar components and the ground substance of human skin derma fixed in formaldehyde showed good preservation of the fibrillar structures. The typical changes in fixed collagen fibrils in negative staining include the lack of transversal lines in the zone A and poor manifestation of microfibrillarity in the zone B. Alterations in the ground substance are more significant. Staining with rutenium red reveals no reticular structure here, but floccular formations appear in the amorphous interfibrillar substance. Other species of rutenium-positive structures (sheaths of elastic fibers and collagen fibrils as well as lines of thin cross-striation of the latter) are well preserved. All these alterations should be taken into consideration in ultramicroscopical examinations of formalin-fixed connective tissue.     

Pulmonary hypoplasia in the connective tissue growth factor (Ctgf) null mouse.

Connective tissue growth factor (CTGF) is a mediator of growth factor activity, and Ctgf knockouts die at birth from respiratory failure due to skeletal dysplasia. Previous microarray analysis revealed Ctgf down-regulation in the hypoplastic lungs of amyogenic mouse embryos. This study, therefore, examined pulmonary development in Ctgf-/- mouse fetuses to investigate if respiration could also have been impaired by lung abnormalities. The Ctgf-/- lungs were hypoplastic, with reduced cell proliferation and increased apoptosis. PDGF-B, its receptor and IGF-I, were markedly attenuated and the TTF-1 gradient lost. Type II pneumocyte differentiation was perturbed, the cells depicting excessive glycogen retention and diminished lamellar body and nuclear size, though able to synthesize surfactant-associated protein. However, type I pneumocyte differentiation was not affected by Ctgf deletion. Our findings indicate that the absence of Ctgf and/or its protein product, CTGF, may induce pulmonary hypoplasia by both disrupting basic lung developmental processes and restricting thoracic expansion.     

多囊卵巢综合征模型大鼠卵巢中结缔组织生长因子及基质金属蛋白酶9的表达

背景：有研究表明,卵泡发育、成熟、排卵和黄体形成以及退化等卵巢的生理过程都涉及细胞外基质的形成,而基质金属蛋白酶9和结缔组织生长因子与细胞外基质的形成有着密切的关系。 目的：探讨卵巢中结缔组织生长因子及基质金属蛋白酶9的表达与多囊卵巢综合征的关系。 方法：将有规律动情周期的SD雌性大鼠随机分为2组,模型组大鼠采用来曲唑灌胃建立多囊卵巢综合征模型,对照组灌胃等量的羧甲基纤维素。当模型组大鼠动情周期固有规律不存在,连续出现间期时采集大鼠的血清和卵巢组织,对照组在其间期采集标本进行检测。 结果与结论：放射免疫分析法和免疫组织化学染色法检测显示,与对照组相比,模型组的血清促黄体生成激素、血清垂体泌乳素、窦前卵泡卵泡膜胞浆中的结缔组织生长因子、卵泡基底膜胞浆结缔组织生长因子、白膜结缔组织生长因子以及黄体中基质金属蛋白酶9表达量明显升高(P < 0.05),促卵泡激素、血清雌二醇和卵泡基底膜中的基质金属蛋白酶9表达量则明显降低(P < 0.05)。结果证实,结缔组织生长因子可能与多囊卵巢综合征中可能与小卵泡的过多生长和排卵障碍有关,基质金属蛋白酶9可能与多囊卵巢综合征中的排卵障碍有关。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

Restricted expression of galactose/N-acetylgalactosamine-specific macrophage C-type lectin to connective tissue and to metastatic lesions in mouse lung.

We investigated expression of macrophage galactose/N-acetylgalactosamine-specific C-type lectin (MMGL) in normal mouse lung tissue and in lungs with metastatic nodules produced by OV2944-HM-1 mouse metastatic ovarian tumour cells. Cells expressing MMGL were detected in tissue sections using a rat monoclonal antibody (mAB) specific for MMGL, mAb LOM-14. The regions containing cells immunostained using mAb LOM-14 were restricted to the connective tissue surrounding blood vessels and respiratory epithelia, whereas alveolar regions of lung parenchyma were essentially devoid of these cells. In contrast, a significant number of cells in the alveolar regions was shown to express Mac-1 antigen (CD11b/CD18) and leucocyte common antigen (CD45). Immunoelectron microscopic study revealed the presence of MMGL in the intracellular vesicles of cells residing in connective tissue. In the tumour-bearing mice, MMGL-positive cells were also present within metastatic nodules. Their localization outside of the nodules was restricted to connective tissue. Cells with Mac-1 antigens were seen both in the nodules and in the alveolar regions. These results indicate that MMGL serves as a unique macrophage marker in mouse lung tissue due to its topographical site-dependent pattern of expression. The present results also suggest a possible involvement of macrophages expressing MMGL in the immune response directed against metastatic tumour cells.