干扰干细胞离子通道对鹿茸形态发生的影响  

Influence of Interfering Ion Channels of Antler Stem Cells on Morphogenesis of Deer Antlers (Cervus nippon)

作　　者：赵海平[1] 褚文辉[1] 刘振[1] 张伟[1] 谷海军[1] 李春义[1] 

机构地区：[1]中国农业科学院特产研究所/特种经济动物分子生物学国家重点实验室/吉林省鹿茸工程研究中心,长春130112

出　　处：《农业生物技术学报》2017年第7期1146-1153,共8页Journal of Agricultural Biotechnology

基　　金：吉林省重点科技攻关项目(No.20150204071NY)

摘　　要：近年来,生物电编码动物/动物器官形态发生成为生命科学研究领域的一个里程碑式成果。迄今为止对该领域的研究仅局限于低等动物。为了初步探索哺乳动物器官的形态发生与生物电的相关性。本研究以梅花鹿(Cervus nippon)鹿茸为模型,选择进入青春期前的1岁龄仔鹿(鹿茸尚未开始发育),雌性和雄性仔鹿各3头;通过外科手术摘取鹿茸形态发生原胚生茸区骨膜(antlerogenic periosteum,AP)组织共10片,利用杜尔伯科改良伊格尔培养基(Dulbecco's modified eagle medium,DMEM)培养基(含10%胎牛血清和2%的青霉素-链霉素)进行离体培养;分别将培养液中加入4种细胞离子通道干扰药物(ivermectin,MS-222,concanamycin A和SCH-28080)对AP组织进行了干扰;连续培养2.5 d。结果发现,不管是使用去极化药物还是超极化药物处理均对鹿茸的形态发育产生了一定程度的影响。为了验证药物的作用效果,本研究对AP细胞进行了离体培养,并选用MS-222作为代表对AP细胞进行了处理。通过Coro Na染色,发现MS-222降低了AP细胞的细胞内钠离子的浓度;通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5)-dimethylthiahiazo(-z-y1)-2,5-di-phenytetrazoliumromide,MTT)实验发现,MS-222没有影响AP细胞的增殖率。说明MS-222之类的药物对AP细胞并不具有细胞毒性,因而其抑制鹿茸发育的最可能的途径是通过对细胞离子通道的干扰影响了AP组织的极化状态。本研究结果表明,AP组织离子通道的状态与鹿茸的发育之间可能存在直接关系。因此推测,鹿茸可能与低等动物模型一样,只有特定的生物电状态才能维持正常的形态发生。本研究为进一步验证生物电编码哺乳动物器官形态发生选定的动物模型提供了理论依据。The theory of bioelectric code is a remarkable achievement for life sciences. It is unknown whether this theory could apply to mammals. In the present study, sika deer （Cervus nippon） antlers were used as a model to investigate if morphogenesis of mammalian organs was also related to bioelectricity. Deer antlers are complex mammalian organs. Morphogenetic primordia of deer antlers is antlerogenic periosteum （AP）. Deer antler morphogenesis can be altered artificially by manipulating AP conveniently. Ten pieces of APs were collected through surgery from 3 male and 3 female sika deer calves before antler generation. These APs were cultured in sterile Dulbecco＇s modified eagle medium （DMEM） medium （containing 10% fetal calf serum （FBS） and 2% penicillin-streptomycin） in vitro and treated with four kinds of drugs for 2.5 d respectively. These drugs were ivermectin, MS-222, concanamycin A and SCH-28080 which were known to be able to alter bioelectricity state of tissues/cells through interrupting ion channels. Before the culture, APs were punctuated with a beam of needles to facilitate the drugs to diffuse into the tissue. After the culture, APs were implanted back to their original places. Subsequent development of antlers was observed weekly and photographed when necessary. Two months later, following results were obtained. Irrespective of the depolarized or hyperpolarized status, antler development was affected. The results from male deer were consistent with those from female deer （mechanical stimulation could induce female deer to grow antlers）. The role of mechanical stimulation was impaired by drug treatment in female deer in this particular case. For the purpose of testing the effect of drug, AP cells were isolated from AP tissue and cultured in sterile DMEM medium （containing 10% FBS and 2% penicillin-streptomycin） in vitro. MS-222 was selected as a representative drug and added to DMED medium. Three groups of AP cells were designed and treated with MS-222 for 12, 24 and 48 h, res

关 键 词：生物电 鹿茸 器官形态发生 

分 类 号：S825[农业科学—畜牧学]