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Cellular distribution of the new growth factor Pleiotrophin (HB-GAM) mRNA in developing and adult rat tissues 总被引:7,自引:0,他引:7
J. M. Vanderwinden P. Mailleux S. N. Schiffmann J. J. Vanderhaeghen 《Anatomy and embryology》1992,186(4):387-406
Summary Pleiotrophin (PTN), also known as HBGAM, belongs to an emerging cytokine family unrelated to other growth factors. We report here the first comprehensive study using in situ hybridization on the cellular distribution of this new heparin-binding growth factor mRNA in rat tissues. PTN mRNA was developmentally expressed in many — but not all — neuroectodermal and mesodermal lineages, whilst no PTN mRNA was detected in endoderm, ectoderm and trophoblast. PTN mRNA was found in the nervous system throughout development, with a post-natal peak of expression. In the adult nervous system, significant expression persisted in hippocampal CA1 pyramidal neurons and in cortical neurons, but also in different non-neuronal cells types in various locations (olfactory nerve, cerebellar astrocytes, pituicytes, Schwann cells surrounding the neurons in sensory ganglia). PTN mRNA was also found during development in the mesenchyme of lung, gut, kidney and reproductive tract, in bone and cartilage progenitors, in dental pulp, in myoblasts, and in several other sites. Expression was differently regulated in each location, but usually faded around birth. In the adult, PTN mRNA was still present in the meninges, the iris, the Leydig cells of the testis and in the uterus. PTN mRNA was also strongly expressed in the basal layers of the tongue epithelium, which is the only epithelium and ectodermal derivative to express PTN mRNA, and this only after birth. PTN is known to be a growth factor for perinatal brain neurons and a mitogen for fibroblasts in vitro. Recently, trophic effects on epithelial cells and a role as a tumour growth factor have been reported. The mechanisms of regulation and the functions of PTN are however still uncertain. Its expression pattern during development suggests important roles in growth and differentiation. Moreover, the presence of PTN mRNA in several adult tissues and the up-regulation of PTN mRNA expression in the gravid uterus indicate that PTN also has physiological functions during adulthood. 相似文献
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Isolation of a unique hepatic stellate cell population expressing integrin α8 from embryonic mouse livers
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Tomohiro Ogawa Yuchang Li Ingrid Lua Andrea Hartner Kinji Asahina 《Developmental dynamics》2018,247(6):867-881
Results: Using antibodies against ALCAM and PDPN, we separated mesothelial cells (MCs) and HSCs from developing livers and identified integrin α8 (ITGA8) as a marker for embryonic desmin+ HSCs that are preferentially localized near the developing liver surface and α‐smooth muscle actin+ perivascular mesenchymal cells around the vein. A cell lineage–tracing study revealed that upon differentiation, MC‐derived HSCs or perivascular mesenchymal cells express ITGA8 during liver development. Using anti‐ITGA8 antibodies, we succeeded in isolating MC‐derived HSCs and perivascular mesenchymal cells from embryonic livers. In direct co‐culture, ITGA8+ mesenchymal cells promoted the expression of hepatocyte and cholangiocyte markers in hepatoblasts. In the normal adult liver, expression of ITGA8 was restricted to portal fibroblasts in the portal triad. Upon liver injury, myofibroblasts increased the expression of ITGA8. 相似文献
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Activation, proliferation, and differentiation of progenitor cells into hepatocytes in the D-galactosamine model of liver regeneration. 总被引:9,自引:19,他引:9
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Rat liver regeneration was studied from 24 hours to 8 days after a single intraperitoneal injection of D-galactosamine (GalN). Morphological changes in the liver were analyzed in parallel with sequential changes in expression of histone-3 mRNA (a marker of cell proliferation), fetal alpha-fetoprotein (AFP) mRNA and gamma-glutamyl transpeptidase (GGT) (markers of fetal hepatocytes), and albumin mRNA and glucose-6-phosphatase (G6Pase) (markers of adult hepatocytes). Proliferation of nonparenchymal epithelial cells (NPC), detected in situ by [3H]thymidine labeling or histone-3 mRNA expression, began after 24 hours primarily in the portal area around the bile ducts. After 2 days, histone-3 labelling intensity increased in rows and clusters of NPC which expanded from the portal zone and invaded into the parenchyma. On days 3 and 5, NPC expressing his-3 mRNA expanded further, forming pseudo-ducts and islet-like structures (NPC structures). Proliferating NPC were positive for GGT. Some GGT positive cells were also positive for the fetal form of AFP mRNA, which lagged behind GGT by 24 hours and peaked on day 5. On day 3, some cells with the appearance of NPC expressed albumin mRNA. Double label in situ hybridization for fetal AFP and albumin mRNAs and dual histochemistry for GGT and G6Pase showed simultaneous expression of these markers in NPC on day 5. Other cells expressing fetal AFP mRNA or GGT on day 5 had a morphological appearance between NPC and hepatocytes (transitional cells). Proliferation of hepatocytes began on day 2, reached maximum on day 5 and then declined. Proliferating hepatocytes did not express fetal AFP mRNA or GGT. These findings indicate that after GalN injury, the liver responds by activation of progenitor cells that proliferate and then differentiate into mature hepatocytes. Adult hepatocytes can also proliferate after GAlN injury, but these hepatocytes do not undergo dedifferentiation/redifferentiation during regeneration of the hepatic lobule. 相似文献
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背景:Nanog、Oct4和Sox2通过调节胚胎干细胞的基因转录,对其多潜能性和自我更新的能力具有关键性的调控作用,脐带间充质干细胞中这些胚胎干细胞相关转录因子的表达情况如何还不太清楚。
目的:研究脐带间充质干细胞中Nanog、Oct4和Sox2等这些胚胎干细胞相关转录因子的表达情况。
方法:胶原酶和胰酶消化法培养脐带间充质干细胞;mTeSRTM1体系进行无滋养层培养人胚胎干细胞,定量PCR比较上述两种细胞中Nanog、Oct4和Sox2 mRNA表达量的差异;免疫荧光检测上述两种细胞中Nanog、Oct4和Sox2的表达情况。
结果与结论:间充质干细胞表达胚胎干细胞标记Nanog、Oct4和Sox2,但Oct4主要表达在胞浆,且以Oct4B为主。脐带间充质干细胞Nanog、Oct4A和Sox2的表达量明显低于胚胎干细胞,其mRNA表达量分别为胚胎干细胞的20%,0.3%,10%左右。通过了解两种细胞Nanog、Oct4和Sox2的表达差异,可为优化脐带间充质干细胞重编程提供依据,也为进一步研究胚胎干细胞相关转录因子在成体干细胞表达起何种作用提供参考。 相似文献
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Y J Wan L Wang T C Wu 《Laboratory investigation; a journal of technical methods and pathology》1992,66(5):646-651
The expression of retinoic acid receptor alpha, beta, and gamma mRNA was examined in developing rat livers and rat hepatoma-derived cell lines H-4-II-E, McA-RH 7777, and 8994 that represent different hepatocyte phenotypes. Northern blot hybridization demonstrated that all three receptor mRNAs were expressed in the fetal livers of different gestational ages, and the levels of expression increased significantly 3 to 4 weeks after birth. In the hepatoma cell lines, the expression pattern of retinoic acid receptor alpha and gamma mRNA did not correlate with the phenotype. In contrast, retinoic acid receptor beta mRNA was only detected in the adult phenotypic H-4-II-E cells but not in McA-RH 7777 and 8994 cells, which represent embryonic and fetal hepatocyte phenotypes, respectively. The levels of retinoic acid receptor beta mRNA in hepatoma cell lines were lower than adult rat liver. These data suggest that the increased expression of retinoic acid receptor beta gene is associated with differentiation or maturation of rat hepatocytes. The effect of retinoic acid on retinoic acid receptor gene expression was also studied in hepatoma cells. Retinoic acid did not regulate retinoic acid receptor gene expression in McA-RH 7777 and 8994 cells, and the retinoic acid receptor beta gene remained inactivated in these cells. However, Southern blot hybridization indicated that the gross structure of retinoic acid receptor beta gene was not altered during malignant transformation. In H-4-II-E cells, retinoic acid increased the expression of retinoic acid receptor beta and gamma gene. Because of the similarity between H-4-II-E cells and normal adult hepatocytes, this type of autoregulation may be a mechanism by which retinoic acid regulates its own effect in vivo. 相似文献
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目的:探讨PDGF-B反义寡脱氧核苷酸(ASODN)对大鼠肝星状细胞(HSCs)增殖及Ⅰ型胶原合成的影响。方法:合成了特异性的PDGF-B硫代磷酸ASODN及其对照错义寡脱氧核苷酸(MSODN),并将其加入至培养的HSCs。采用MTT法检测PDGF-B ASODN对HSCs的增殖抑制作用,RT-PCR检测PDGF-B和collagen-Ⅰ mRNA的表达,流式细胞仪与ELISA法分别检测PDGF-B的表达和Ⅰ型胶原的合成。 结果:PDGF-B ASODN在终浓度为10 μmol/L,作用HSCs-T6细胞48 h时,能明显地抑制其增殖,降低PDGF-B和collagen-ⅠmRNA的表达;FCM与ELISA分析表明,HSCs表达PDGF-B和合成Ⅰ型胶原都明显低于对照组,而对照组寡脱氧核苷酸在相同浓度下未见抑制效应。结论:PDGF-B ASODN可以抑制HSCs的增殖、Ⅰ型胶原的合成、内源性PDGF-B的表达,可能成为一种有用的抗肝纤维化基因治疗的药物。 相似文献
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目的:探讨实验性肝硬化细胞生长激素受体的表达变化。方法:采用硫代乙酰胺腹腔注射复制大鼠肝硬化模型,采用放射配体法、RT-PCR、图像分析检测不同阶段肝硬化肝组织和肝硬化肝细胞中生长激素受体及其mRNA的表达水平,并分别与正常肝组织和正常肝细胞比较。结果:大鼠肝硬化肝组织表达生长激素受体,其表达数量明显少于正常肝组织,并且随着肝硬化的进展、肝组织胶原纤维相对含量的增加而进一步减少;大鼠肝硬化肝细胞生长激素受体的位点数量明显少于正常肝细胞,大鼠肝硬化肝组织生长激素受体mRNA的表达量也明显少于正常肝组织。结论:大鼠肝硬化肝细胞表达生长激素受体,其表达水平降低;肝硬化越严重,该表达减少越明显。大鼠肝硬化肝组织生长激素受体表达下调的机制可能是肝硬化肝组织生长激素受体mRNA的表达明显减少。 相似文献
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目的:探讨骨髓间质干细胞(MSCs)对活化态肝星状细胞(HSCs)增殖的影响。 方法: 分别从骨髓和肝脏分离纯化培养大鼠MSCs及HSCs,塑料板传代培养激活HSCs;在半透膜(transwell insert)上接种MSCs,在6孔塑料培养板上接种HSCs,建立上下双层细胞共培养体系;大鼠正常肝细胞系(BRLs)及HSCs培养分别作为对照。免疫细胞化学检测平滑肌激动蛋白(α-SMA)与结蛋白的表达,IBAS 2.5软件分析阳性染色表达量。 结果: HSCs与MSCs共培养24 h,HSCs表现轻度增殖抑制,随着培养时间延长,HSCs增殖活性受抑制更明显,在48 h和72 h的抑制率分别达15.7%与30.3%,与BRLs共培养体系比较有显著差异;与MSCs共培养72 h, HSCs表达α-SMA量明显低于两个对照组BRLs及HSCs培养体系(50.2% vs 90.2%、95.6%, P<0.01);而结蛋白表达的量在3组共培养体系中均无显著差异。 结论: MSCs具有分泌细胞因子抑制HSCs增殖活性的潜能,在治疗肝纤维化中可能发挥作用。 相似文献
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神经生长因子低亲合力受体在肝纤维化患者和大鼠肝星状细胞的表达 总被引:3,自引:0,他引:3
目的探讨神经生长因子低亲合力受体(P75)在肝纤维化患者和大鼠肝星状细胞(HSCs)的分布及作用机制。方法对四氯化碳(CCL)法制备的肝纤维化大鼠肝组织及肝穿刺获取的肝纤维化患者肝组织,常规石蜡包埋;大鼠离体培养的活化HSCs离心涂片,行免疫组织化学染色,确定P75的表达分布。结果P75在大鼠离体培养的活化HSCs膜,免疫组织化学染色呈阳性,在肝纤维化患者HSCs膜和肝细胞膜,免疫组织化学染色呈阳性,在肝纤维化大鼠HSCs膜和肝细胞膜,免疫组织化学染色呈阳性,在健康人和正常对照大鼠HSCs膜,免疫组织化学染色呈阴性,表明人和大鼠活化的HSCs膜表达P75。结论P75为肝纤维化的治疗提供了新的靶点。 相似文献
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Transforming growth factor-beta and platelet-derived growth factor signal via c-Jun N-terminal kinase-dependent Smad2/3 phosphorylation in rat hepatic stellate cells after acute liver injury 总被引:2,自引:0,他引:2
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Yoshida K Matsuzaki K Mori S Tahashi Y Yamagata H Furukawa F Seki T Nishizawa M Fujisawa J Okazaki K 《The American journal of pathology》2005,166(4):1029-1039
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Interactions of vimentin- or desmin-expressing liver cells with N-acetylglucosamine-bearing polymers
It is necessary to develop highly functionalized liver cell culture systems for liver tissue engineering such as bioartificial livers and liver cell chips. To maintain a high level of hepatocyte function, well-organized patterning culture systems of hepatocytes and nonparenchymal cells would be advantageous. To design the patterning culture system using these cells, cell-recognizable polymers should be useful to regulate not only the hepatocytes, but also the nonparenchymal cells. Here, we report that N-acetylglucosamine (GlcNAc)-bearing polymers are useful as nonparenchymal cell-recognizable polymers. It has previously been reported that mesenchymal cells adhered to GlcNAc-bearing polymer-coated dishes through surface vimentin. It was also observed that nonparenchymal cells expressing vimentin or desmin specifically adhered to GlcNAc-bearing polymer-coated dishes. Especially, in hepatic stellate cells (HSCs) cultured on GlcNAc-bearing polymer-coated dishes, the expression of α-smooth muscle actin as an activated HSCs marker was suppressed in long-term. Therefore, HSCs were shown to maintain a quiescent state on PVGlcNAc-coated dishes during a long-term culture. These results demonstrated that GlcNAc-bearing polymers could be beneficial to culture nonparenchymal cells such as HSCs. Our findings suggest that galactose- and GlcNAc-bearing polymers can regulate the culture of all liver cells and may be useful tools for the establishment of liver tissue engineering. 相似文献
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Long-term maintenance of hematopoietic stem cells does not require contact with embryo-derived stromal cells in cocultures 总被引:5,自引:0,他引:5
Oostendorp RA Robin C Steinhoff C Marz S Bräuer R Nuber UA Dzierzak EA Peschel C 《Stem cells (Dayton, Ohio)》2005,23(6):842-851
We recently established that two midgestation-derived stromal clones--UG26-1B6, urogenital ridge-derived, and EL08-1D2, embryonic liver-derived--support the maintenance of murine adult bone marrow and human cord blood hematopoietic repopulating stem cells (HSCs). In this study, we investigate whether direct HSC-stroma contact is required for this stem cell maintenance. Adult bone marrow ckit+ Ly-6C- side population (K6-SP) cells and stromal cells were cocultured under contact or noncontact conditions. These experiments showed that HSCs were maintained for at least 4 weeks in culture and that direct contact between HSCs and stromal cells was not required. To find out which factors might be involved in HSC maintenance, we compared the gene expression profile of EL08-1D2 and UG26-1B6 with four HSC-nonsupportive clones. We found that EL08-1D2 and UG26-1B6 both expressed 21 genes at a higher level, including the putative secreted factors fibroblast growth factor-7, insulin-like growth factor-binding proteins 3 and 4, pleiotrophin, pentaxin-related, and thrombospondin 2, whereas 11 genes, including GPX-3 and HSP27, were expressed at a lower level. In summary, we show for the first time long-term maintenance of adult bone marrow HSCs in stroma noncontact cultures and identify some secreted molecules that may be involved in this support. 相似文献
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Adult hepatocytes and liver-cell progenitors play a role in restoring liver tissue after injury. For the study of progenitor cells in liver repair, experimental models included (a) surgical removal of liver tissue by partial hepatectomy; (b) acute injury by carbontetrachloride; (c) acute injury by d-galactosamine (GalN) and N-nitrosomorpholine (NNM); and (d) chemical hepatocarcinogenesis by feeding NNM in low and high doses. Serological and immunohistological detection of alpha-fetoprotein gene expression served to follow pathways of cellular differentiation. Stem cells were not required in models of surgical removal of parenchyma and in carbon tetrachloride intoxication of adult hepatocytes. In contrast, regeneration of liver occurred through biliary epithelial cells in injuries induced by GalN and NNM. These biliary epithelial cells, collectively called oval cells, are most probably derived from the canals of Hering. Proliferating bile duct cells reached a level of differentiation with reactivation of foetal genes and significant alpha-1-fetoprotein (AFP) synthesis signalling a certain degree of retrodifferentiation with potential stemness. Due to the same embryonic origin of bile ducts and hepatocytes, biliary epithelium and its proliferating progeny (oval cells) have a defined role in liver regeneration as a transit and amplification compartment. In their early proliferation stage, oval cells were heavily engaged in DNA synthesis ([3H]thymidine labelling). Pulse-chase experiments during experimental hepatocarcinogenesis exhibited their development into hepatocytes with high risk for transformation and leading to foci of altered hepatocytes. Hepatocellular carcinomas may arise either from proliferating/differentiating oval cells or from adult hepatocytes; both cell types have stem-like properties. AFP-positive and AFP-negative carcinomas occurred in the same liver. They may represent random clonal origin. The heterogeneity of phenotypic marker (AFP) correlated with a process of retrodifferentiation. 相似文献
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Yoshihisa Fujikura Hidehiko Ohta Toshio Hirai Tetsuo Fukumoto 《Anatomical record (Hoboken, N.J. : 2007)》1993,235(3):335-341
Four monoclonal antibodies were raised against crude gap junction fractions of rat liver to clarify the distribution of gap junctions during animal development and to analyze gap junction expression in vivo and the polarity of hepatocytes in vitro. Among the monoclonal antibodies obtained, HAM8 antibody recognized the 27-kDa rat liver gap junction protein connexin 32. This antibody recognized gap junctions at the contiguous faces of hepatocytes, and the antigen was also observed in exocrine pancreas and salivary gland but not in kidney, heart, esophagus, or thymus. HAM8 did not react with amphibian or fish liver, heart, esophagus, stomach, or intestine as assessed via the immunofluorescence method on frozen sections. A few hepatocytes and many hemopoietic cells were seen in rat fetal liver at 15 days of gestation. HAM8 antigen was expressed on some hepatocytes but not on any hemopoietic cells. As the fetus grew, the number of hepatocytes in the liver increased gradually, together with the amount of HAM8 antigen. The distribution of HAM8 antigen at 25 days after birth was similar to that in adult liver. When the expression of HAM8 antigen was examined in primary cultured hepatocytes using the immunofluorescence method, the antigen was observed clearly between the hepatocytes. However, most of the HAM8 antigen on the free surface of hepatocytes disappeared within 4 hr. HAM8 antigen was not expressed on AH7974 rat hepatoma cells when they formed small islets in the rat peritoneal cavity or within the liver. When HAM8 IgG antibody was injected intravenously, the HAM8 signal was expressed in the liver. © 1993 Wiley-Liss, Inc. 相似文献