cgVEGF164基因对小鼠毛囊生长的影响

血管内皮生长因子(vascular endothelial growth factor, VEGF)是一种二聚体糖蛋白,能够诱导毛囊血管内皮细胞的增殖和迁移,调节毛囊周围毛细血管生成,进而影响毛囊的生长发育。已知cgVEGF164是绒山羊VEGF-A基因的一种主要剪接变体,但cgVEGF164基因是否具有调控毛囊生长发育的作用目前尚不清楚。为初步探究cgVEGF164基因对毛囊生长的影响及其机制,本研究通过原核显微注射制备cgVEGF164转基因过量表达小鼠。通过HE染色法比较转基因小鼠和非转基因对照小鼠毛囊的直径和密度,利用WesternBlot检测小鼠背部皮肤中信号蛋白ERK1/2、AKT1、LEF1的磷酸化水平。本研究成功获得5只阳性cg VEGF164转基因小鼠(雌雄比为4:1,阳性率8.5%);与非转基因对照小鼠相比,转基因小鼠毛囊直径增大、密度增加;经检测转基因小鼠中ERK1/2、AKT1、LEF1的磷酸化水平均上调。结果表明,cgVEGF164基因具有促进小鼠毛囊生长的作用,推测该功能可能与cgVEGF164影响ERK1/2、AKT1和LEF1等信号蛋白的磷酸化有关。     

The effect of cold after whole-body irradiation of the mouse

Mice were placed in a cold environment (4 °C) directly after whole-body irradiation. Those irradiated with a lethal dose showed higher lethality than mice irradiated with the same dose but placed in room temperature. The response was also altered after irradiation with a sublethal dose. At various periods after irradiation mice were injected with¹²⁵IUdR, the tissue uptake of which is an index of DNA synthesis. The result showed that cold treatment after irradiation caused slower cell renewal in the spleen and bone marrow, but that the thymus was only marginally affected. Furthermore, the concentrations of erythrocytes in the peripheral blood reached a lower level in the cold-treated group. Finally, the levels of the thyroid hormones T3 and T4 in the blood were measured and it was found that the T3/T4 ratio was higher in the cold-treated mice. It is suggested that during prolonged exposure to cold after irradiation the cell recovery in the haemopoietic system is exposed to hormonal action that induces significant alterations in the postirradiation cell kinetics.The animal experiments described herein were approved by the Regional Research Ethical Committee according to the Swedish National Laws SFS 1988:539 and LSFS 1989:41     

Cell death (apoptosis) in hair follicles and consequent changes in the width of hairs after irradiation of growing follicles

Irradiation of anagen (growing) hair follicles results in a dose-dependent increase in the number of histologically identifiable fragments of dead cells (apoptotic fragments). The incidence of apoptotic fragments is linearly related to dose, increasing at a rate of 2.92 fragments per follicle section per Gy. The effects of doses of 0.2 Gy can be easily detected. Subjective attempts to associate clusters of fragments with dead or dying cells suggests that the number of fragments per cell increases with dose (about 1.7 fragments per cell after 1 Gy to about 2.7 fragments per cell after 5 Gy). There is a natural incidence of cell death in controls (0.13 +/- 0.06 fragments per follicle section with about 1.4 fragments per dead or dying cell). Damage to the follicle cells is expressed in the differentiated product of the follicle, the hair, by a reduction in width. This is probably the cellular basis for the production of dysplastic hairs. The hair width has been measured and is reduced by about 7 per cent for every gray of radiation. The value of the hair and hair follicles as potential biological dosimeters is discussed.     

Changes after irradiation in the number of mitotic cells and apoptotic fragments in growing mouse hair follicles and in the width of their hairs

The hair follicle or its differentiated product, the hair, which represents the linear historical record of the follicular proliferative activity, could provide a biological dosimeter of value for dose distribution determinations after accidental exposure. Here we present some further studies on irradiated mouse hair follicles and hair, and discuss the difficulties in obtaining similar data for humans. The incidence of cell death in the follicles has been shown elsewhere to be maximum 12 h after irradiation, and it increases with dose. Here we confirm that doses of 0.2-0.4 Gy can be readily detected. We show here that there is only a little more cell death in the larger follicles even though they contain many more cells and mitotic figures. About one-third of all the dead cell fragments in a follicle can be seen in a good longitudinal follicle section. Mitotic activity declines progressively with dose in the large follicles, which start with more mitotic cells, showing the dose-dependent changes most readily. The dead cells are morphologically identical to apoptotic cells at the level of the light microscope, and they fragment into several bodies, the number of which increases with dose. The total number of apoptotic bodies or fragments in whole large follicles increases almost 100-fold over a range of 1.3 Gy (from 0.2 to 1.5 Gy) and about tenfold over the range 0.2-0.5 Gy. The estimated number of dead (apoptotic) cells increases about sevenfold over the same 1.3-Gy range. The width of the middle portion of the broadest, awl, hairs measured 12 days after irradiation decreases with increasing dose. About 80% of the hairs show an obvious reduction in width after 2 Gy and the effects of a dose of about 1 Gy can be detected. The width of the hair is reduced by 10-14% per Gy. A comparison has been made between BDF1 (black) and BALB-c (albino) mice. The large follicles contain similar numbers of mitotic cells, but the BALB-c mice are more sensitive both in terms of the radiation-induced apoptosis and in terms of a reduction in awl hair width.     

Hoxa4 expression in developing mouse hair follicles and skin

We have examined the expression of the Hoxa4 gene in embryonic vibrissae and developing and cycling postnatal pelage hair follicles by digoxigenin-based in situ hybridization. Hoxa4 expression is first seen in E13.5 vibrissae throughout the follicle placode. From E15.5 to E18.5 its expression is restricted to Henle's layer of the inner root sheath. Postnatally, Hoxa4 expression is observed at all stages of developing pelage follicles, from P0 to P4. Sites of expression include both inner and outer root sheaths, matrix cells, and the interfollicular epidermis. Hoxa4 is not expressed in hair follicles after P4. Hoxb4, however, is expressed both in developing follicles at P2 and in catagen at P19, suggesting differential expression of these two paralogous genes in the hair follicle cycle.     

Effects of gamma-radiation on the differentiation of mouse melanocytes in the hair follicles

Pregnant mice were whole-body irradiated with a single acute dose of gamma-rays (60Co) to investigate the effect of gamma-radiation on embryonic melanoblasts. The effect was studied by scoring changes in the differentiation of melanocytes in the hair follicles of mice heterozygous for the recessive coat color mutation pink-eyed dilution (p). Abnormal round melanocytes were found in the hair matrix and the dermal papilla of F1 offspring 3.5 days after birth. However, these round melanocytes possessed a melanin deposition of similar intensity to normal hair follicular melanocytes. The frequency of the abnormal hair follicles increased in a dose-dependent manner. Moreover, higher frequencies were found in the animals irradiated at earlier stages of embryonic development. These results indicate that gamma-radiation affects dendritogenesis and the location of mouse melanocytes in the hair follicles, with greater effects seen at the earlier stages of development.     

The mutant gene "wal" is active in cells of mouse hair follicles

In order to determine the place of action of the mutant gene waved alopecia (wal), we have obtained chimeric wal/wal c/c Gpi-1aa<-->+/+ C/C Gpi-1bb animals by aggregation of eight-cellular embryos of BALB/c-wal/wal mice and CBA (+/+) mice. The presence or absence of the chimeric structure was determined from the mosaic nature of fur color and hair structure, as well as on the basis of the presence of electrophoretically distinct variants of glucosephosphate isomerase in blood. Chimeras had alternating transverse patches of different lengths and widths consisting of curly (genotype wal/wal) or straight (genotype +/+) hairs. The percentage of cells with wal/wal mutant genotype in chimeras established on the basis of glucosephosphate isomerase isozymes varied from 10 to 80%. A higher percentage of the parental wal/wal component in chimeras correlated with the number of patches having wavy hairs. Analysis of the fur pattern represented by the alternation of transverse patches of wavy or straight hairs in chimeric wal/wal (+/+ mice has shown that mutant gene wal acts in ectodermal cells of hair follicles.     

Effects of X-irradiation on rat hair follicles

The alterations of normal growth of hair follicles following irradiation of rat tails with X-rays have been analysed. The changes in follicle length are attributed to a series of deleterious events occurring in the hair matrix after irradiation. A dose-dependent reduction of length was demonstrated for a given post-irradiation interval. An approximate exponential response was obtained for all conditions excepting low doses for which a marked reduction of follicle length was observed a short time after irradiation and was ascribed to the depletion of highly radiosensitive cells. At later post-irradiation times this response was obscured by the resumption of follicle growth.     

Late effects in the mouse small intestine after a clinically relevant multifractionated radiation treatment

Late radiation effects were investigated in the mouse small intestine after a daily fractionated radiation treatment. Mice were given 14 X 3 Gy in 2 weeks over a partial abdominal irradiation field. There was evidence for late injury in the intestinal epithelium, the submucosa, and the subserosa. Late damage in the epithelium was shown histologically by a reduced crypt number and villus atrophy at 3 and 6 months but not at 24 h after the end of treatment. The reduction in crypt number was significant in the ileum at 3 and 6 months after irradiation: 100 +/- 4 and 98 +/- 5 (SEM) per circumference, respectively, versus 132 +/- 3 and 146 +/- 6 in age-matched controls (P less than 0.01, t test). The mitotic activity in the crypts of the irradiated animals was significantly increased at all investigated times, suggesting a prolonged but insufficient compensatory response to maintain the mucosal integrity. The repercussion on intestinal epithelial function was, at least in part, reflected by a progressively reduced body weight gain up to 5 g at 3 months after treatment. The ability of the surviving crypt stem cells to form microcolonies after irradiation, however, was not impaired. Evidence for injury in the submucosa was provided from macroscopic and histological examination. Macroscopically, at 6 months after treatment, narrowed and rigid bowel segments surrounded by fibrotic adhesions were observed, causing partial intestinal obstruction. In addition, sometimes focal areas of hemorrhage and infarction in small bowel segments were present. Histologically, diffuse and pronounced submucosal edema without increased fibrosis was seen, together with markedly dilated small blood vessels in focal areas of macroscopic intestinal infarction. The intestinal perfusion, as assessed by 86Rb extraction, was significantly but transiently reduced at 3 months after irradiation. These data suggest mainly late effects in the small intestine after this daily fractionated irradiation treatment. The reduced number of epithelial cells and the submucosal edema are possibly mediated by radiation injury in the intestinal microvasculature.     

The chemokine SDF-1/CXCL12 regulates the migration of melanocyte progenitors in mouse hair follicles

Mouse skin melanocytes originate from the neural crest and subsequently invade the epidermis and migrate into the hair follicles (HF) where they proliferate and differentiate. Here we demonstrate a role for the chemokine SDF-1/CXCL12 and its receptor CXCR4 in regulating the migration and positioning of melanoblasts during HF formation and cycling. CXCR4 expression by melanoblasts was upregulated during the anagen phase of the HF cycle. CXCR4-expressing cells in the HF also expressed the stem cell markers nestin and LEX, the neural crest marker SOX10 and the cell proliferation marker PCNA. SDF-1 was widely expressed along the path taken by migrating CXCR4-expressing cells in the outer root sheath (ORS), suggesting that SDF-1-mediated signaling might be required for the migration of CXCR4 cells. Skin sections from CXCR4-deficient mice, and skin explants treated with the CXCR4 antagonist AMD3100, contained melanoblasts abnormally concentrated in the epidermis, consistent with a defect in their migration. SDF-1 acted as a chemoattractant for FACS-sorted cells isolated from the anagen skin of CXCR4–EGFP transgenic mice in vitro, and AMD3100 inhibited the SDF-1-induced migratory response. Together, these data demonstrate an important role for SDF-1/CXCR4 signaling in directing the migration and positioning of melanoblasts in the HF.     

Expression of prostaglandin E(2) receptor subtypes in mouse hair follicles.

We investigated the mRNA distribution of the prostaglandin (PG) E(2) receptor subtypes and cyclooxygenases (COXs) in hair follicles of the mouse dorsal skin. In the 3-week hair follicles, which are in the anagen phase, EP3 and EP4 mRNA were expressed in the dermal papilla cells and the outer root sheath cells located in the hair bulb region, respectively. In the 8-week hair follicles, which are in the telogen phase, the signals for both EP3 and EP4 mRNAs had disappeared. To study the hair cycle-dependent expression of mRNAs for the EPs and COXs, an area of dorsal hair was depilated from 8-week-old mice. On days 8 and 12 after depilation, EP3 and EP4 mRNA were reexpressed in the dermal papilla cells and the outer root sheath cells, and the induction of COX-2 mRNA was also observed in the outer root sheath cells, the upper area of EP4 expression site. These results suggest that EP3 and EP4 receptors may involve in the development and regrowth of the hair follicles.     

毛囊分子调控机制

毛囊是产生毛被的组织,具有周期性,经历生长期、衰退期和休止期三个时期,毛囊与皮脂腺组成毛囊皮脂腺单位。本文从分子层面分别对调控毛囊周期性变化、毛囊皮脂腺发育以及毛囊中影响毛被光泽度及毡结度的因子等给予了详细阐述,这些均为提高山羊绒产量和质量提供了重要思路。     

The kinetics of repair in mouse lung after fractionated irradiation

The kinetics of repair of sublethal damage in mouse lung was studied after fractionated doses of 137Cs gamma-rays. A wide range of doses per fraction (1.7-12 Gy) was given with interfraction intervals ranging from 0.5 to 24 h. The data were analysed by a direct method of analysis using the incomplete repair model. The half-time of repair (T1/2) was 0.76 h for the pneumonitis phase of damage (up to 8 months) and 0.65 h for the later phase of damage up to 12 months. The rate of repair was dependent on fraction size for both phases of lung damage and was faster after large dose fractions than after small fractions. The T1/2 was 0.6 h (95 per cent c.1. 0.53, 0.69) for doses per fraction greater than 5 Gy and 0.83 h (95 per cent c.1 0.76, 0.92) for doses per fraction of 2 Gy. Repair was nearly complete by 6 h, at least for the pneumonitis phase of damage. To the extent that extrapolation of these data to humans may be valid, these results imply that treatments with multiple fractions per day that involve the lung will not be limited by the necessity for interfraction intervals much longer than 6 h.