Wnt3a在毛囊及黑素细胞谱系中的表达

目的:探讨毛囊周期中,Wnt3a在毛囊及黑素细胞中的表达变化。方法:以DCT-LacZ转基因小鼠为动物模型,通过X-gal染色技术观察黑素细胞谱系在小鼠皮肤中的分布情况;采用X-gal染色结合免疫组化方法检测Wnt3a在毛囊及黑素细胞谱系中的表达情况;采用RT-PCR方法对小鼠皮肤全层Wnt3a和TYR的mRNA表达进行半定量分析。结果:在生长期毛囊中,Wnt3a蛋白在表皮、毛囊外根鞘Bulge区、内根鞘以及毛球部均有表达,在黑素干细胞与黑素细胞也观察到Wnt3a;在退化期,Wnt3a的表达逐渐减弱,仅在外根鞘有较弱的表达,但黑素干细胞中没有观察到Wnt3a;在静止期,几乎检测不到Wnt3a的表达;TYR mRNA与Wnt3a mRNA在毛囊周期中的表达模式一致,在生长期最强,退化期减弱,静止期最弱。结论:Wnt3a可能对黑素细胞谱系分化起到促进作用。     

IGF1和IGF1R在小鼠第一个毛囊生长周期皮肤的表达

毛囊生长周期中,真皮乳头和毛基质间的基质 上皮信号调控细胞的增殖和分化。多功能细胞调控因子胰岛素样生长因子1（IGF1)是该信号路径的成员之一。第1个毛囊生长周期决定着毛囊的正常生长和发育,但IGF1在此期的作用未见报道。实时荧光定量PCR结果显示,IGF1在生长期皮肤中的相对表达量最低,在退化期表达量最高,在静止期表达量又降低。与生长初期相比,IGF1在退化期和静止期的表达量呈差异极显著（P＜0.01）;胰岛素样生长因子1受体（IGF1R）在生长期皮肤中的相对表达量最高,在退化期表达量最低,而在静止期表达量又升高。与生长初期相比,IGF1R在退化期和静止期的表达量呈差异极显著（P＜0.01）。Western 印迹结果显示,IGF1和IGF1R蛋白在小鼠皮肤第1个毛囊生长周期各阶段的表达趋势分别与其mRNA的表达趋势一致;免疫组织化学结果表明,IGF1主要分布在小鼠表皮,而IGF1R免疫阳性在小鼠毛囊毛球部、内外根鞘和毛乳头均有分布。以上实验结果揭示,IGF1和IGF1R在小鼠皮肤第1个毛囊生长周期的各阶段的差异性表达,可能在毛囊生长周期各阶段的转化过程中参与了黑色素的形成。然而,IGF1和IGF1R表达趋势不一致,提示IGF1在小鼠皮肤中发挥作用时,并非只与IGF1R结合才能发挥作用。     

HFSC标记在阿尔巴斯绒山羊毛囊及毛囊干细胞中的表达

旨在检测最广泛应用的毛囊干细胞标记在阿尔巴斯绒山羊毛囊组织中的表达情况,为今后内蒙古绒山羊毛囊组织学和细胞学领域相关研究提供依据。以阿尔巴斯绒山羊为研究对象,对其进行Krt15、Krt19、CD34和Sox9等的冰冻切片和细胞免疫组化实验。以上4个标记均在外根鞘中表达;Krt15、Krt19在隆突部表达,在隆突部下端不连续表达;CD34在隆突部和隆突部下端都有较强的表达;Sox9主要集中在隆突部表达。Krt15、Krt19、CD34和Sox9共同表达的部位为毛囊隆突部,因此可结合表达部位特征选用为阿尔巴斯绒山羊毛囊隆突部来源的毛囊干细胞鉴定标记。     

HGF/c-met在小鼠毛囊生长周期中的表达

目的 HGF及其受体c-met是很多系统中间充质-上皮互相作用的主要的介导者,毛囊周期性生长是典型的间充质-上皮互作模型,为证实HGF/c-met信号在小鼠毛囊周期生长过程中是否发挥作用。方法本实验运用免疫组织化学的方法对HGF/c-met在ICR小鼠毛囊生长期、退化期和休止期进行组织定位。结果 HGF主要位于真毛乳头和皮脂腺,c-met主要位于毛基质、根鞘部和表皮。HGF及其受体在小鼠毛囊生长期表达量达到最高值,在退化期基本不表达,在休止期-生长期过渡时HGF及其受体表达量均上升。结论 HGF/c-met信号在ICR小鼠毛囊生长过程中起调控作用。     

β-catenin在小鼠雄性激素源性脱发模型毛囊分化发育中的作用

[目的]旨在构建雄性激素源性脱发的小鼠毛囊分化发育及周期的模型,并研究雄性激素对雄性激素源性受体(AR)、β-catenin等毛囊发育周期中关键性调控因子在脱发模型小鼠毛囊分化发育中的作用机制的影响。[方法]小鼠皮下注射高浓度睾酮(5 mg/kg)后进行背部皮肤毛发脱毛的同步化处理,通过HE染色观察毛囊发育情况,并利用qPCR、免疫组化检测AR、β-catenin基因和蛋白的表达情况。[结果]睾酮组毛囊首个毛囊兴盛期的发育时间推迟5 d且提前进入休止期,休止期毛囊的毛干与真皮之间间隙增大。睾酮组与对照组相比,AR在毛囊发育早期减弱,第7 d对照组表达下降而睾酮组开始均匀分布于除毛乳头处的整个毛囊。22 d时,对照组毛囊已经进入休止期,AR基本不表达,而睾酮组在外根鞘和连接组织鞘仍可看到有少量表达。睾酮组第5 dβ-catenin在新生的内根鞘、皮质区与毛母质区有较高的表达,随后表达差异与对照组不明显。qPCR结果 AR基因表达与免疫组化趋势基本一致,但雄性激素对β-catenin基因表达的抑制一直持续到了兴盛期后期。[结论]通过建立雄性激素源性脱发的小鼠动物模型,发现雄性激素对毛囊发育分化的作用可能是通过影响AR和β-catenin等毛囊发育关键调控因子来实现的,其具体机制有待进一步研究。     

EdU体外标记毛囊外根鞘干细胞的实验研究

摘要 目的：探寻一种简单有效的追踪定位毛囊干细胞的分裂增殖标记的方法,为后期毛囊干细胞增殖分化迁移机制的研究打下基础。方法：体外无菌条件下获取昆明乳鼠触须毛囊外根鞘,以5 % FCS+MEM作为基础培养液,添加4种不同浓度EdU（5-Ethynyl-2’-deoxyuridine）细胞标记物,进行毛囊外根鞘再生培养,冰冻切片,H.E染色,使用KeyFluor488对EdU所标记的再生细胞进行检测。结果：H.E染色和EdU标记染色结果发现EdU细胞标记物浓度越高,毛囊外根鞘再生形态越差,同时当EdU浓度为20 μ mol/L时,毛囊外根鞘增殖细胞标记效果是最好的。结论：EdU细胞标记物对毛囊外根鞘中的干细胞增殖活性具有一定的毒副作用,可能会影响细胞的正常生理代谢功能,浓度为20 μ mol/L EdU可以有效的标记毛囊干细胞增殖,并对毛囊再生结构影响较小。     

毛囊干细胞

毛囊干细胞被认为是具慢周期性特点,但特定条件下具有较高增殖能力和克隆形成潜能的细胞。它们在形态学和生物化学上处于较原始的状态,常具有多潜能性。利用干细胞标记技术和克隆形成能力检测手段,人们发现毛囊干细胞主要存在于位于毛囊上半部的隆突部位。毛囊干细胞潜在的分子标记包括β1-整合素、α6-整合素、CD71、角蛋白19、p63和CD34,而在体内和体外它们的分子标记也不尽相同。毛囊隆突部位的干细胞能够分化成表皮、上皮性毛根鞘、发杆和皮脂腺。在个体发育过程中,它们具有分化成其他多种细胞系的能力。对于在毛发形态形成和生长周期中毛囊干细胞的行为,研究者们提出了多种假说,包括隆突激活假说和干细胞迁移假说。如今,毛囊干细胞主要应用于制备皮肤的代替品。     

P75NTR在绒山羊毛囊生长周期中的表达

实验旨在研究绒山羊（Capra hircus）毛囊生长相关基因的表达规律,为绒山羊分子育种提供参考。本实验采用RT-PCR、组织免疫荧光、Western blot等方法,研究神经营养素受体P75^NTR在辽宁绒山羊皮肤组织中的表达和分布情况。结果表明：在毛囊生长周期的三个时期,均监测到P75^NTR mRNA及蛋白的存在,P75^NTR的荧光信号在退行期要强于其他两个时期,且在退行期毛囊外根鞘细胞中检测到P75^NTR的高表达。以上结果表明,P75^NTRmRNA及蛋白的表达与毛囊生长周期变化有一定的相关性,P75^NTR受体在绒山羊的毛囊周期性变化中发挥着重要的功能。     

毛囊干细胞标记物的研究进展

毛囊干细胞是一类位于毛囊隆突区的成体干细胞,对毛囊的周期性生长,表皮和皮脂腺的更新以及皮肤损伤后修复有着至关重要的作用。毛囊干细胞的标记物是对毛囊干细胞进行分离和鉴定的重要依据,对毛囊干细胞的基础研究起着关键作用。因此寻找特异性较高的毛囊干细胞标记物成为了近年的研究热点。本文按毛囊干细胞标记物在细胞中所处部位进行分类,将其分为位于细胞膜、细胞质、细胞核的标记物,综述了目前国内外主要采用的整合素、角蛋白、CD34、CD200等标记物以及新发现的Lgr5、Sox9、Tcf3等基因标记物。     

角蛋白15在大鼠皮肤发育中表达的研究

目的研究角蛋白15(K15)在大鼠皮肤发育中的表达状况,定位表皮干细胞.方法以不同年龄大鼠背部皮肤为标本,用组织学方法,观察出生后大鼠皮肤的形态发育变化;以K15单克隆抗体为一抗,进行免疫组织化学染色,观察K15在大鼠皮肤中的表达状况.结果(1)组织学方法显示,随着年龄的增长,大鼠背部表皮细胞层数逐渐变少;在毛囊的生长周期中,以隆突区为界,毛囊上段为恒定区,下段呈周期性变化(2)免疫组化染色显示,毛囊隆突区细胞胞浆表达K15,随年龄的增长,K15阳性细胞出现在毛母质细胞区、毛囊外根鞘和表皮基底层.结论表皮干细胞位于毛囊隆突区,与表皮的更新和毛囊的周期性变化有关.     

小鼠毛囊不同生长周期中MITF下游色素相关基因的定位表达及相关性分析

小眼畸形转录因子（MITF）不仅是黑色素细胞发育、增殖和存活的必要调节因子,而且对调节相关酶和黑素体蛋白表达来确保黑色素产生具有至关重要的作用。MITF下游色素相关基因在小鼠毛囊生长周期中的表达及相关性仍有待研究。HE染色结果表明不同毛囊时期的小鼠毛囊呈现典型的组织形态学结构;免疫组织化学显示,MITF、GPNMB、OA1、TYR、TYRP2在不同毛囊生长周期中的毛基质及内外毛根鞘均有不同程度的阳性表达。黑色素测定结果表明,在毛囊生长初期和中期,碱性可溶性总黑色素（ASM）、真黑素（EM）以及褐黑素（PM）相对含量高于毛囊生长末期。蛋白免疫印迹结果表明,MITF、GPNMB、OA1、TYR、TYRP2在毛囊生长初期和中期蛋白质相对水平明显高于毛囊生长末期。实时荧光定量PCR结果表明, MITF、GPNMB、OA1、TYR、TYRP2、PMEL在毛囊生长初期和中期,mRNA相对表达量显著高于毛囊生长末期。在不同毛囊生长周期小鼠皮肤的MITF下游色素相关基因表达存在显著差异,表明上述因子在维持黑色素细胞色素生成是不可或缺的因素。     

Expression of basement membrane components through morphological changes in the hair growth cycle

The amount and distribution of fibronectin associated with hair follicles was found to vary during the hair growth cycle in the rat. Immunocytochemical staining of follicles in mid-late anagen (the growth stage) revealed the presence of fibronectin in the dermal papilla matrix, in the basement membrane separating this from the epithelial cells of the hair bulb, and in the basement membrane and connective tissue sheath which underly the cells of the outer root sheath. Early in catagen, the transitional stage, staining of the dermal papilla matrix disappeared. Fibronectin persisted in the basement membrane and connective tissue sheath, which undergo corrugation and apparent thickening in catagen. After follicle shortening, the telogen (resting) stage is reached, at which point fibronectin staining was found to be minimal, being restricted to the basement membrane around the secondary germ. The onset of anagen, involving cell division and follicle elongation, was associated with a great increase in the amount of fibronectin in this zone and in and around the dermal papilla. Analysis of entry into anagen by [3H]thymidine incorporation and autoradiography revealed that growth could be detected before the increase in fibronectin expression. However, growing cells, even in a suprabasal position, always had some fibronectin at their surface. Immunoelectron microscopy of early anagen follicles confirmed the light microscopic findings and also showed that fibronectin was present in small vesicles close to the surface of dermal papilla and some epithelial cells. Increased deposition of laminin and type IV collagen in early anagen follicles was also noted, emphasizing the importance of basement membrane components during morphogenetic events in vivo.     

Expression of E6 and E7 papillomavirus oncogenes in the outer root sheath of hair follicles extends the growth phase and bypasses resting at telogen.

Hair follicle growth cycle proceeds through a series of stages in which strict control of cell proliferation, differentiation, and cell death occurs. Transgenic mice expressing human papillomavirus type 16 E6/E7 papillomavirus oncogenes in the outer root sheath (ORS) display a fur phenotype characterized by lower hair density and the ability to regenerate hair much faster than wild-type mice. Regenerating hair follicles of transgenic mice show a longer growth phase (anagen), and although bulb regression (catagen) occurs, rest at telogen was not observed. No abnormalities were detected during the first cycle of hair follicle growth, but by the second cycle, initiation of catagen was delayed, and rest at telogen was again not attained, even in the presence of estradiol, a telogen resting signal. In conclusion, expression of E6/E7 in the ORS delays entrance to catagen and makes cells of the ORS insensitive to telogen resting signals bearing to a continuous hair follicle cycling in transgenic mice.     

Analysis of the expression pattern of involucrin in human scalp skin and hair follicles: hair cycle-associated alterations

Involucrin is a structural component of the keratinocyte cornified envelope that is expressed early in the keratinocyte differentiation process. It is a component of the initial envelope scaffolding and considered as a marker for keratinocyte terminal differentiation. The expression pattern of involucrin in human scalp skin and hair follicle cycle stages is not fully explored. This study addresses this issue and tests the hypothesis that "the expression of involucrin undergoes hair follicle cycle-dependent changes". A total of 50 normal human scalp skin biopsies were examined (healthy females, 51-62?years) using immunofluorescence staining methods and real-time PCR analysis. In each case, 50 hair follicles were analyzed (35, 10 and 5 follicles in anagen, catagen and telogen, respectively). Involucrin was prominently expressed in the human scalp skin and hair follicles, on both gene and protein levels. The protein expression showed hair follicle cycle-associated changes i.e. a very strong expression during early and mature anagen, intermediate to strong expression during catagen and prominent decline in the telogen phase. The expression value of involucrin in both anagen and catagen was statistically significantly higher than that of telogen hair follicles (p?相似文献   

Morphogenesis of the hair follicle during catagen

Summary During catagen, the transition period between growth and quiescence, the growing (anagen) hair follicle is reorganized to form the resting (telogen) follicle. The last portion of the hair shaft produced at the onset of catagen consists only of cortex. Surrounding the cortex and attached tightly to it are the club cells, which resemble the cortex in structure and development except that the filaments of the club are oriented randomly and do not exhibit the keratin pattern seen in the cortex. The club cells in turn are attached to a capsule of germ cells which are formed by progressive transformation of the outer root sheath cells at the middle of the growing follicle. When the capsule of germ cells is formed, the follicle below it undergoes resorption, presumbaly mediated by hydrolytic enzymes. As the follicle disintegrates, the surrounding basal lamina undergoes extensive pleating and is eventually resorbed. Collagen fibers around the basal lamina are engulfed and degraded by the large number of macrophages that surround the hair follicle at this time. The dermal papilla remains as a compact ball of cells just below the capsule of germ cells.This study constitutes publication No. 428 from the Oregon Regional Primate Research Center, supported by Grant No. FR-00163.I wish to thank Mrs. Janice Anderson for patient and excellent technical assistance and Mr. Joel Ito for the drawing.     

Expression of galectin-1 and -3 and of accessible binding sites during murine hair cycle

Although protein-carbohydrate interactions are supposed to play key roles in cell adhesion, signalling and growth control. Their exact role in skin physiology has only recently been investigated. The endogenous lectins galectin-1 and galectin-3 have been identified in skin including hair follicles. Here, we analyzed the expression and distribution of these galectins and their binding sites in C57BL/6 mice during hair cycle. The expression of galectin-1 and galectin-3 binding sites was found to be predominantly hair cycle-dependent showing some overlapping to the expression of galectin-1 and -3. The outer root sheath (ORS) expressed galectin-1 binding sites during anagen IV to VI and in early catagen, whereas galectin-1 was expressed from early anagen to late catagen. The ORS expressed galectin-3 binding sites during catagen transition corresponding to a galectin-3 expression during anagen V and catagen. The innermost layer of the ORS expressed galectin-3 binding sites during anagen VI until catagen VIII, but galectin-3 during anagen III to IV and catagen. The inner root sheath (IRS) expressed galectin-3 binding sites only in anagen IV but missed expression of any of the two galectins. The matrix cells expressed galectin-3 binding sites in catagen II-III as well as galectin-3 during anagen V to catagen IV. The present study provides the first evidence for a cycle-related expression of both galectin-1 and -3 and their binding sites during murine hair cycle.     

Intercellular adhesion molecule-1 and hair follicle regression.

Although the intercellular adhesion molecule-1 (ICAM-1) is recognized for its pivotal role in inflammation and immune responses, its role in developmental systems, such as the cyclic growth (anagen) and regression (catagen) of the hair follicle, remains to be explored. Here we demonstrate that ICAM-1 expression in murine skin is even more widespread and more developmentally regulated than was previously believed. In addition to endothelial cells, selected epidermal and follicular keratinocyte subpopulations, as well as interfollicular fibroblasts, express ICAM-1. Murine hair follicles express ICAM-1 only late during morphogenesis. Thereafter, morphologically identical follicles markedly differ in their ICAM-1 expression patterns, which become strikingly hair cycle-dependent in both intra- and extrafollicular skin compartments. Minimal ICAM-1 and leukocyte function-associated (LFA-1) protein and mRNA expression is observed during early anagen and maximal expression during late anagen and catagen. Keratinocytes of the distal outer root sheath, fibroblasts of the perifollicular connective tissue sheath, and perifollicular blood vessels exhibit maximal ICAM-1 immunoreactivity during catagen, which corresponds to changes of LFA-1 expression on perifollicular macrophages. Finally, ICAM-1-deficient mice display significant catagen acceleration compared to wild-type controls. Therefore, ICAM-1 upregulation is not limited to pathological situations but is also important for skin and hair follicle remodeling. Collectively, this suggests a new and apparently nonimmunological function for ICAM-1-related signaling in cutaneous biology.