Cataractogenesis in neonatal Sprague-Dawley rats by N-methyl-N-nitrosourea

Cataract was induced by a single intraperitoneal injection of 100 mg/kg N-methyl-N-nitrosourea (MNU) to 0-, 5-, 10-, 15-, or 20-day-old male and female Sprague-Dawley rats. In day 0, 5, 10, and 15 MNU-treated rats, mature cataracts were constantly seen 7, 14, 14, and 30 days after dosing, respectively. In the day 20 MNU-treated rats, only subcapsular cataract was seen 30 days after dosing. Therefore, the rats exposed to MNU at an earlier age caused cataract more rapidly and severely. In the day 0 MNU-treated rats, 7-methyldeoxyguanosine DNA adduct was detected in the lens epithelial nuclei 12 hours after MNU dosing, followed by apoptosis, which was confirmed by morphology, by TUNEL signals, and by DNA ladder and peaked 3 days after MNU dosing. In the apoptosis cascade, upregulation of Bax, downregulation of Bcl-2, and increased CPP32 protease (caspase-3) activity were seen 12 hours after MNU dosing. Therefore, the pathogenesis of MNU-induced cataract was associated with DNA adduct formation in the lens epithelial cell nuclei leading to apoptosis by upregulation of Bax protein, downmodulation of Bcl-2 protein, and activation of caspase-3.     

Induction of hair regrowth in the alopecia site of IFN-gamma knockout mice by allografting and IFN-gamma injection into the transplantation site.

We previously demonstrated that around 6 weeks of age most of the interferon-gamma (IFN-gamma)(-/-) but none of the IFN-gamma(+/+) C57BL/6 mice began to lose hair in their dorsal or occipital areas or both and that a single s.c. injection of IFN-gamma into IFN-gamma(-/-) mice at 3 but not at 8 weeks of age (or later) could protect all the mice from alopecia. Here, we report hair regrowth in the alopecia site of IFN-gamma(-/-) mice at 8 weeks of age (or later) by the combination of IFN-gamma and allografting. Skin or tumor allografting and IFN-gamma injections into the transplantation site induced hair regrowth in the alopecia site of IFN-gamma(-/-) mice at 8-66 weeks of age, whereas IFN-gamma injections into the hairless site or allografting alone was ineffective in causing the hair regrowth. Histologic findings showed that the hair cycle in the region of alopecia of IFN-gamma(-/-) mice was blocked at the anagen stage and that in the IFN-gamma(-/-) mice treated with IFN-gamma and allografting, the cycle was at the telogen stage. The therapeutic effects were maintained for >1 year.     

Alopecia of IFN-gamma knockout mouse as a model for disturbance of the hair cycle: a unique arrest of the hair cycle at the anagen phase accompanied by mitosis.

Interferon-gamma(-/-) (IFN-gamma(-/-)) and IFN-gamma(+/+) C57BL/6 mice (3 weeks of age) completed the production of morphogenesis-derived hair. Around 6 weeks of age, however, most of the IFN-gamma(-/-) but none of the IFN-gamma(+/+) mice began to lose hairs in the dorsal and occipital areas in the absence of inflammatory reactions, and the alopecia was sustained for at least several 10-week periods of observation. A single subcutaneous injection of IFN-gamma to IFN-gamma(-/-) mice at 3, but not 4, 5, or 8 weeks of age could protect all the mice from alopecia, revealing that the lack of IFN-gamma around 3 weeks of age is directly responsible for the alopecia. Histologic features showed that the hair follicles of the IFN-gamma(+/+) mice passed through the anagen (4-5 weeks of age) and catagen/telogen ( approximately 6 weeks of age) phases, whereas those of IFN-gamma(-/-) mice (5 weeks of age or older) stayed in the anagen phase. TUNEL and bromodeoxyuridine experiments suggested that an arrest with unlimited DNA synthesis of the hair cycle in the anagen phase by the lack of IFN-gamma-dependent apoptosis in the midfollicle region and diffuse shedding of previously formed hair induced alopecia in IFN-gamma(-/-) mice.     

Induction of fetal malformations after treatment of mouse embryos with methylnitrosourea at the preimplantation stages.

The effects of methylnitrosourea (MNU) on the development of preimplantation mouse embryos were investigated in this study. ICR mice were treated intraperitoneally with single doses of 10, 20, and 30 mg MNU/kg body wt on day 0, 1, 2, or 3 of pregnancy. The uterine contents were examined on day 18 of pregnancy. The fetuses were examined for external and skeletal abnormalities. No significant differences were observed in the number of implantation sites between all the MNU-treated groups and controls. MNU treatment on day 2 or 3 of pregnancy caused dose-dependent significant increases in the incidence of abnormal fetuses over the control level, while treatment on day 0 or 1 failed to cause an increase of abnormalities. Cleft palate, exencephalus, and malformed vertebrae were the most common types of abnormalities. In the embryo transfer experiments, the frequency of fetal abnormalities induced when embryos were transferred from MNU-treated females to untreated pseudopregnant females was significantly higher than that induced when embryos were transferred from untreated females to MNU-treated or untreated pseudopregnant females. The results in the present study confirm and extend the previously proposed hypothesis that the direct effects of MNU on preimplantation embryos make a significant contribution to the induction of fetal malformations.     

Keratin 17 modulates hair follicle cycling in a TNFalpha-dependent fashion

Mammalian hair follicles cycle between stages of rapid growth (anagen) and metabolic quiescence (telogen) throughout life. Transition from anagen to telogen involves an intermediate stage, catagen, consisting of a swift, apoptosis-driven involution of the lower half of the follicle. How catagen is coordinated, and spares the progenitor cells needed for anagen re-entry, is poorly understood. Keratin 17 (K17)-null mice develop alopecia in the first week post-birth, correlating with hair shaft fragility and untimely apoptosis in the hair bulb. Here we show that this abnormal apoptosis reflects premature entry into catagen. Of the proapoptotic challenges tested, K17-null skin keratinocytes in primary culture are selectively more sensitive to TNFalpha. K17 interacts with TNF receptor 1 (TNFR1)-associated death domain protein (TRADD), a death adaptor essential for TNFR1-dependent signal relay, suggesting a functional link between this keratin and TNFalpha signaling. The activity of NF-kappaB, a downstream target of TNFalpha, is increased in K17-null skin. We also find that TNFalpha is required for a timely anagen-catagen transition in mouse pelage follicles, and that its ablation partially rescues the hair cycling defect of K17-null mice. These findings identify K17 and TNFalpha as two novel and interdependent regulators of hair cycling.     

Rapid induction of cataract by a single intraperitoneal administration of N-methyl-N-nitrosourea in 15-day-old Sprague-Dawley (Jcl: SD) rats.

Cataract was induced by a single intraperitoneal injection of N-methyl-N-nitrosourea (MNU) in 15-day-old Sprague-Dawley (Jcl: SD) rats. The threshold dose of MNU required for cataract induction in a 3-4 week time period was 70 mg/kg; 60 mg/kg was ineffective. Males and females were both equally affected. Mature cataract as confirmed histologically by degeneration, swelling, vacuolation, liquefaction of the lens fibers, and formation of Morgagni-like water vacuoles was seen in 80% (8/10), 70% (7/10) and 90% (9/10) of 100, 80 and 70 mg/kg MNU-treated rats, respectively, 4 weeks after dosing (43 days of age). At this time point, lens epithelial apoptosis was only rarely seen, but proliferating cell nuclear antigen (PCNA) labeled atypical nuclei with vacuolated lens fibers and macrophage migration were present within the injured lens. Cataracts were the only lesions induced by MNU and there were no other intra- or extraocular lesions seen. The dosing and timing schedule for the MNU administration in rats used in the present study is effective in rapidly causing cataract to occur.     

Time-specific action of N-methyl-N-nitrosourea in the occurrence of retinal dysplasia and retinal degeneration in neonatal mice

The morphologic response of neonatal mouse retina to the alkylating agent N -methyl- N -nitrosourea (MNU) was examined at different periods of retinal development. A dose of 60 mg/kg N -methyl- N -nitrosourea was injected intraperitoneally to neonatal C₅₇BL mice at 0, 3, 5, 8, 11, 14, 17, and 20 days of age and to C₃H mice at 0 days of age, and the retinas were examined sequentially. In the C₆₇BL mice, MNU evoked a time-dependent occurrence of retinal dysplasia and retinal degeneration. With MNU treatment at day 0 and day 3 (the stage of retinal cell proliferation), retinal dysplasia characterized by the progressive disorganization of neuroblasts, which led to the formation of rosettes, was found in the outer neuroblastic/nuclear layer above the normal pigment epithelial cells during days 8–20, but decreased at day 50. The rosettes were surrounded by photoreceptor segments and Muller cell processes, and by photoreceptor nuclei. The MNU response was related to retinal differentiation; following MNU treatment at day 5 or 8 (the stage of retinal cell differentiation) the cells were much less sensitive (i.e. no retinal response was found). However, with MNU treatment at days 11, 14, 17, and 20 (after cellular differentiation), retinal degeneration characterized by selective photoreceptor apoptosis was seen. These results suggest that there is a critical period for the time of MNU administration in the development of mouse retinal lesions. In C₃H ( rd/rd ) mice, MNU treatment at day 0 resulted in retinal degeneration with only slight rosette formation at the peripheral retina.     

Hair growth modulation by topical immunophilin ligands: induction of anagen, inhibition of massive catagen development, and relative protection from chemotherapy-induced alopecia.

Selected immunophilin ligands (IPLs) are not only potent immunosuppressants but also modulate hair growth. Their considerable side effects, however, justify at best topical applications of these drugs for the management of clinical hair growth disorders. Therefore, we have explored hair growth manipulation by topical cyclosporin A (CsA) and FK 506 in previously established murine models that mimic premature hair follicle regression (catagen) or chemotherapy-induced alopecia, two major pathomechanisms underlying human hair loss. We confirm that topical CsA and FK 506 induce active hair growth (anagen) in the back skin of C57BL/6 mice with all follicles in the resting stage (telogen) and show that both IPLs also inhibit massive, dexamethasone-induced, premature catagen development in these mice. Furthermore, we demonstrate that CsA and FK 506 provide relative protection from alopecia and follicle dystrophy induced by cyclophosphamide, possibly by favoring the dystrophic anagen pathway of follicle response to chemical damage. Although it remains to be established whether these IPLs exert the same effects on human hair follicles, our study provides proof of the principle that topical IPLs can act as potent manipulators of clinically relevant hair-cycling pathomechanisms. This strongly encourages one to explore the use of topical IPLs in the management of human hair growth disorders.     

Adenovirus-Mediated Wnt5a Expression Inhibits the Telogen-To-Anagen Transition of Hair Follicles in Mice

The canonical Wnt/β-catenin pathway plays an important role in hair cycle induction. Wnt5a is a non-canonical Wnt family member that generally antagonizes canonical Wnt signaling in other systems. In hair follicles, Wnt5a and canonical Wnt are both expressed in cells in the telogen stage. Wnt5a has been shown to be critical for controlling hair cell fate. However, the role that Wnt5a plays in the transition from the telogen to anagen stage is unknown. In this study, using whole-mount in situ hybridization, we show that Wnt5a is produced by several other cell types, excluding dermal papilla cells, throughout the hair cycle. For example, Wnt5a is expressed in bulge and secondary hair germ cells in the telogen stage. Our studies focused on the depilated 8-week-old mouse as a synchronized model of hair growth. Interestingly, overexpression of adenovirus Wnt5a in the dorsal skin of mice led to the elongation of the telogen stage and inhibition of the initiation of the anagen stage. However, following an extended period of time, four pelage hair types grew from hairless skin that was induced by Wnt5a, and the structure of these new hair shafts was normal. Using microarray analysis and quantitative arrays, we showed that the expression of β-catenin and some target genes of canonical Wnt signaling decreased after Wnt5a treatment. These data demonstrate that Wnt5a may inhibit the telogen stage to maintain a quiescent state of the hair follicle.     

Bone morphogenetic protein signaling regulates postnatal hair follicle differentiation and cycling

Hair follicle morphogenesis and cycling were examined in transgenic mice that overexpress the bone morphogenetic protein (BMP) inhibitor Noggin under the control of the neuron-specific enolase promoter. The Noggin transgene was misexpressed in the proximal portion of the hair follicle, primarily the matrix cells, apart from the usual expression in neurons. Transgene expression appeared only after induction of both the primary (tylotrich) and secondary (nontylotrich) pelage hair follicles had already occurred, thus allowing examination of the role of BMP signaling in follicles that had been induced normally in the presence of BMPs. The overexpression of Noggin in these animals resulted in a dramatic loss of hair postnatally. There was an apparently normal, but shortened period of postnatal hair follicle morphogenesis, followed by premature initiation of hair follicle cycling via entry into the first catagen transformation. This resulted in a complete loss of hair shafts from the nontylotrich hair follicles in these mice while the tylotrich hair follicles were normal. The onset of anagen of the first postnatal hair follicle cycle was also accelerated in the transgenic mice. Our results show that BMP signaling is specifically required for proper proliferation and differentiation during late morphogenesis of nontylotrich hair follicles and that inhibition of this signaling pathway may be one of the triggers for the onset of catagen when the follicles are in anagen and the onset of anagen when the follicles are in telogen. Ectopic sebocyte differentiation was another hallmark of the phenotype of these transgenic mice suggesting that BMP signaling may be an important determinant of lineage selection by common progenitor cells in the skin. BMPs likely promote a hair follicle-type differentiation pathway of keratinocytes while suppressing the sebaceous differentiation pathway of skin epithelium.     

β-catenin与小鼠毛囊衰老性变化相关

 摘要：目的探讨β-catenin与毛囊衰老性变化的关系。方法 取老龄与年轻小鼠处于毛囊周期各阶段的皮肤,RT-PCR和Western blot检测β-catenin在mRNA和蛋白水平的表达; IF检测比较β-catenin蛋白表达在毛囊中的组织学分布。结果 老年鼠背皮的β-catenin表达相较处于同一毛囊周期阶段的年轻鼠皮肤在mRNA水平和蛋白质水平均有上调,出生后34个月的小鼠生长期皮肤中β-catenin蛋白的表达为0.9024±0.0167,显著高于出生后35天的0.8012±0.0184。 出生后34个月的小鼠静止期皮肤中β-catenin蛋白表达为0.6348±0.0241,显著高于出生后23天的0.6348±0.0241（P＜0.05）。IF检测显示β-catenin的表达集中在外根鞘和Bulge细胞,也可以观察到β-catenin的入核现象。结论β-catenin表达量上调,周期节律不明显,表达部位不同于年轻鼠可能是衰老相关性白发产生的原因之一。     

毛发生长周期对小鼠皮肤创伤愈合的影响

目的　探讨处于不同毛发生长周期的C57BL/6小鼠皮肤创伤愈合速度。 方法　制备小鼠皮肤创伤模型,计算术后0、3、7 d创面愈合率,愈合率=（原始创面面积－未愈合的创面面积）/原始创面面积×100%,比较毛发静止期（Hair telogen stages）小鼠和毛发生长期（Hair anagen stages）小鼠伤口愈合速度。采用HE染色比较伤口愈合的组织形态结构差异,利用BrdU检测伤口周围细胞增殖。 结果　毛发生长期的小鼠皮肤伤口愈合率显著高于毛发静止期的小鼠伤口愈合率。HE染色显示毛发生长期小鼠伤口周围表皮细胞层较多,且表皮细胞向伤口迁移增强;BrdU检测显示毛发生长期小鼠皮肤伤口周围表皮BrdU+ 细胞数多于毛发静止期小鼠。 结论　毛发生长期的小鼠皮肤创伤愈合率高于毛发静止期小鼠,这一结果为进一步探讨毛囊在创伤愈合过程中的作用提供研究基础,也为选择皮肤创伤愈合动物模型提供指导。     

Stress inhibits hair growth in mice by induction of premature catagen development and deleterious perifollicular inflammatory events via neuropeptide substance P-dependent pathways

It has been much disputed whether or not stress can cause hair loss (telogen effluvium) in a clinically relevant manner. Despite the paramount psychosocial importance of hair in human society, this central, yet enigmatic and controversial problem of clinically applied stress research has not been systematically studied in appropriate animal models. We now show that psychoemotional stress indeed alters actual hair follicle (HF) cycling in vivo, ie, prematurely terminates the normal duration of active hair growth (anagen) in mice. Further, inflammatory events deleterious to the HF are present in the HF environment of stressed mice (perifollicular macrophage cluster, excessive mast cell activation). This provides the first solid pathophysiological mechanism for how stress may actually cause telogen effluvium, ie, by hair cycle manipulation and neuroimmunological events that combine to terminate anagen. Furthermore, we show that most of these hair growth-inhibitory effects of stress can be reproduced by the proteotypic stress-related neuropeptide substance P in nonstressed mice, and can be counteracted effectively by co-administration of a specific substance P receptor antagonist in stressed mice. This offers the first convincing rationale how stress-induced hair loss in men may be pharmacologically managed effectively.     

Mechanisms of photoreceptor cell apoptosis induced by N-methyl-N-nitrosourea in Sprague-Dawley rats

A single intraperitoneal injection of 75 mg/kg N-methyl-N-nitrosourea (MNU) was given to 50-day-old female Sprague-Dawley rats and examined sequentially 12 and 24 hours, and 3 and 7 days after MNU treatment. Photoreceptor cell death was evoked in all treated rats. After MNU treatment, 7-methyldeoxyguanosine DNA adduct was detected selectively in photoreceptor cell nuclei at 12 hours, followed by photoreceptor cell apoptosis as confirmed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling signals which peaked at 24 hours and continued until day 7 when several layers of photoreceptor cell nuclei were left. In apoptosis cascade, down-regulation of Bcl-2 was seen at 12 hours and up-regulation of Bax was seen at 24 hours, and caspase family (caspase 3/CPP32, caspase 6/Mch2, and caspase 8/FLICE protease) activities peaked 72 hours after MNU treatment. Therefore MNU-induced photoreceptor cell death was attributed to DNA adduct formation restricted to photoreceptor cell nuclei leading to photoreceptor cell apoptosis by up-regulation of Bax protein, down-modulation of Bcl-2 protein, and activation of caspases 3, 6, and 8.     

Keratinocyte growth factor is an important endogenous mediator of hair follicle growth, development, and differentiation. Normalization of the nu/nu follicular differentiation defect and amelioration of chemotherapy-induced alopecia.

The growth and development of hair follicles is influenced by a number of different growth factors and cytokines, particularly members of the fibroblast growth factor (FGF) family. Keratinocyte growth factor (KGF or FGF-7) is a recently identified 28-kd member of the FGF family that induces proliferation of a wide variety of epithelial cells, including keratinocytes within the epidermis and dermal adnexa. Because KGF induces marked proliferation of keratinocytes, and both KGF and KGF receptor (KGFR) mRNA are expressed at high levels in skin, we sought to localize KGF and KGFR in skin by in situ hybridization. KGFR mRNA was relatively strongly expressed by keratinocytes in the basilar epidermis as well as throughout developing hair follicles of rat embryos and neonates. KGF mRNA was expressed at lower levels than was KGFR but could be localized to follicular dermal papillae in rat embryos and neonates. These results prompted us to investigate the effects of KGF on hair follicles in two distinct murine models of alopecia. In the first model, recombinant KGF (rKGF) induced dose-dependent hair growth over most of the body in nu/nu athymic nude mice when administered intraperitoneally or subcutaneously over 17 to 18 days. When administered subcutaneously, rKGF induced the most extensive hair growth at the sites of injection. Histologically, rKGF induced marked follicular and sebaceous gland hypertrophy, a normalization of the nu/nu follicular keratinization defect, and an increase in follicular keratinocyte proliferation as assessed by bromodeoxyuridine labeling. In the second model, a neonatal rat model of cytosine arabinoside chemotherapy-induced alopecia in which interleukin-1, epidermal growth factor, and acidic FGF have all demonstrated some degree of alopecia cytoprotection, rKGF induced a dose-dependent cytoprotective effect, abrogating as much as 50% of the alopecia in this model when administered beginning 1 day before the onset of chemotherapy. Taken together, these data suggest that KGF is an important endogenous mediator of normal hair follicle growth, development, and differentiation.     

黑视素基因转染给光双极细胞部分恢复MNU诱导的视网膜感光细胞变性小鼠的视觉

 目的： 探索定向转染内源性光感受蛋白黑视素（melanopsin/opsin 4,Opn4）基因进入给光型双极细胞后,视网膜变性小鼠模型中视网膜神经元的光反应以及整体视觉行为的改变。方法：选用由甲基亚硝基脲（N-methyl-N-nitrosourea,MNU）诱导的成年CD1小鼠作为视网膜变性模型。于P0~P1 CD1乳鼠视网膜底注射Grm6-Opn4-GFP质粒,Grm6-GFP作为阴性对照。通过电转进行基因转染。术后5周对基因转染小鼠腹腔注射MNU诱导视网膜感光细胞变性,对照组注射生理盐水,共设计5个实验组：正常对照组（normal）、生理盐水Grm6-Opn4-GFP对照组（NS+melanopsin）、MNU诱导模型Grm6-Opn4-GFP治疗组（MNU+melanopsin）、MNU诱导模型Grm6-GFP对照组（MNU+GFP）和MNU诱导组（MNU）。诱导后连续7 d进行明暗箱测试,统计动物在暗箱中的活动时间比。随后进行视网膜电图（electroretinogram,ERG）测试,计算体现给光双极细胞光反应的b波峰值、潜伏期和反映视网膜神经节细胞（retinal ganglion cells,RGCs）光反应的明视负波反应（photopic negative response,PhNR）。利用免疫荧光法检测动物视网膜黑视素基因转导效果。结果：黑视素可以被定向转染进入视网膜给光双极细胞。明暗箱实验显示MNU诱导7 d后Grm6-Opn4-GFP转染的CD1小鼠滞留在黑箱的时间显著长于未转染组（P<0.05）,ERG测试显示Grm6-Opn4-GFP转染的CD1小鼠的b波也有明显恢复（P<0.05）。结论：定向转染内源性光感受蛋白黑视素基因进入给光型双极细胞可部分恢复视网膜变性模型动物视觉。     

Hair growth pattern in nude mice

Nude mice are not bald but instead show an 'abortive' reduced hair growth on different sites of the integument. An albino (NMRI-nu) and a pigmented (C57BL/6-nu) strain of nude mice were examined as to whether the regional distribution pattern of this anagen hair proliferation is subject to the same ontogenetic development as in hairy mice. Hairy mice of both strains served as a comparison. Hair distribution was documented macroscopically by drawing and photography in a total of 415 mice of both sexes up to 421 days of age. Because of the pigmentation of the growing anagen hair follicles, the growth areas in the pigmented nude mice were distinctly visible whereas in the albino mice they were roughly recognisable from the boundaries of hair covering. The regional distribution of the 'abortive' anagen hair pattern in both nude strains corresponded to the wave-like course of the adult hair generations of hairy mice. As in older hairy mice, the hair cycle duration in nude mice was prolonged from an age of 121-180 days, the hair growth areas appeared reduced and less symmetrically orientated. Differences of up to 33% in body mass between the lighter nude and +/nu mice made ontogenetic comparison impossible so that all information is based on direct pattern or age comparison. The significance of experiments on the skin and hair follicles of nude mice is further increased if litters are examined comparatively and the temporal and spatial dimension of the follicle proliferation is considered more carefully than has been the case until now.     

去卵巢和性成熟期ICR雌性小鼠动情周期的观察

 目的: 观察处于性成熟期和去卵巢ICR小鼠的动情周期特点。方法: ICR雌性小鼠分为正常对照组、假手术组和去卵巢组,术后第8天开始,连续12 d,每日9:00和16:00分别做2次阴道涂片,记录并观察各组小鼠外阴和阴道涂片细胞特征,规律动情周期发生率,动情周期天数和动情周期各期持续时间。结果: 正常对照组与假手术组动情周期特点表现相似,85%处于性成熟期的ICR小鼠呈规律动情周期,多数为6 d周期(83%),少数为5 d动情周期(17%);与5 d相比,6 d的动情期持续时间明显增加(P<0.05)。5 d和6 d动情周期阴道涂片细胞特征在数量和种类略有所不同。去卵巢组小鼠约76%呈动情周期紊乱,表现为连续动情间期。结论: ICR雌性小鼠具有自身品系的动情周期特点。     

Oral phenytoin protects against experimental cyclophosphamide-chemotherapy induced hair loss

In the current study, effects of oral phenytoin on hair growth in cyclophosphamide-treated rats were assessed with the goal of evaluating the ability of phenytoin to suppress chemotherapy-induced hair loss. Thirty-six rats were randomly assigned to six groups (1k6) of six each (n = 6). In all groups, anagen was induced in flank skin of rats by depilation. On day 9 (anagen VI), rats were injected once with either distilled water (groups 1–3) or cyclophoshamide (groups 4–6). From day 10, rats in group 1 and 4 received oral vehicle (distilled water), groups 2 and 5 received oral phenytoin (50 mg/kg), while groups 3 and 6 also received oral phenytoin (100 mg/kg). Drug or vehicle was administered daily for a period of 28 days. The flank area was serially photographed. At the end of the experimental period, rats were sacrificed to correlate visible hair growth with a histological profile of follicle response and recovery. Glutathione (GSH), glutathione peroxidase (GPX) and lipid peroxidation status were assessed. Cyclophosphamide (CYP) treatment was associated with gross morphologic and histological evidence of hair loss in the flanks, microscopic evidence of hair-shaft thinning, increased skin lipid peroxidation, decreased GSH level, and reduction in GPX activities. Phenytoin co-administration was associated with evidence of improved hair growth, increased hair-shaft thickness, reduced skin lipid peroxidation, increased GSH level, and increased GPX activities. This study showed that oral phenytoin can suppress hair loss due to CYP therapy in rats; however, further studies are needed to evaluate its potential application in chemotherapy-induced alopecia.     

Alopecia in mice infected with murine cytomegalovirus (MCMV).

When mouse cytomegalovirus was injected subcutaneously into 4-12 day old CDI mice there was infection of dermal cells and the dermal papillae of hair follicles. Infected cells were never seen in the epidermis nor in the epithelium of hair follicles. When larger doses of virus (5 X 10(4) pfu) were given, dermal infection led to gross necrosis of the skin, ulceration, scabbing and healing with alopecia. Smaller doses (10(4) pfu) did not cause gross necrosis but damage to follicles resulted in alopecia or sparse hair growth. Skin lesions were not seen after infection of 4-8 week old mice, even when the inoculated skin area had been epilated, or when hyaluronidase was mixed with the virus inoculum. These experiments show that cytomegalovirus, in contrast to herpes simplex and varicella-zoster viruses, infects dermal but not epidermal cells, and that dermal tropism is age-restricted.