湖南师范大学自然科学学报
湖南師範大學自然科學學報
호남사범대학자연과학학보
ACTA SCIENTIARUM NATURALIUM UNIVERSITATIS NORMALIS HUNANENSIS
2013年
4期
63-68
,共6页
李玉珑%农小献%李虹辉%王贝贝%刘希良%褚武英%张建社%李琦华
李玉瓏%農小獻%李虹輝%王貝貝%劉希良%褚武英%張建社%李琦華
리옥롱%농소헌%리홍휘%왕패패%류희량%저무영%장건사%리기화
鳜鱼%生肌调节因子%MyoD%整体胚胎原位杂交%发育表达
鱖魚%生肌調節因子%MyoD%整體胚胎原位雜交%髮育錶達
궐어%생기조절인자%MyoD%정체배태원위잡교%발육표체
Siniperca chuatsi%myogenic regulatory factors%MyoD%whole-mount in situ hybridization%devel-opmental expression
生肌调节因子是肌肉形成的关键控制因素。通过RT-PCR方法克隆得到鳜鱼主要生肌调节因子MyoD序列。鳜鱼MyoD基因cDNA长度为813 bp，编码270个氨基酸，推导的氨基酸序列含有1个bHLH结构。经与其他动物MyoD氨基酸序列比对发现不同动物MyoD的bHLH结构域保守性高。通过合成地高辛标记的反义RNA探针对鳜鱼不同发育阶段的胚胎进行整体胚胎原位杂交，结果发现：MyoD最早在肌节早期被检测到信号。伴随着鳜鱼体节从头部往后发育，MyoD也从头部往尾部表达。在体节期，MyoD在中后部表达。尾芽期MyoD的表达量达到最大值。然而，尾芽期过后，头部MyoD的表达量急剧减少，但尾部仍然有强烈的信号。在血流期仍然能在鳜鱼尾部检测到MyoD的微弱信号。到孵出期在鳜鱼鱼眼的后方和胸部腹侧检测到MyoD的强信号，而身体其他地方检测不到信号。仔鱼期没有检测到MyoD表达的信号。
生肌調節因子是肌肉形成的關鍵控製因素。通過RT-PCR方法剋隆得到鱖魚主要生肌調節因子MyoD序列。鱖魚MyoD基因cDNA長度為813 bp，編碼270箇氨基痠，推導的氨基痠序列含有1箇bHLH結構。經與其他動物MyoD氨基痠序列比對髮現不同動物MyoD的bHLH結構域保守性高。通過閤成地高辛標記的反義RNA探針對鱖魚不同髮育階段的胚胎進行整體胚胎原位雜交，結果髮現：MyoD最早在肌節早期被檢測到信號。伴隨著鱖魚體節從頭部往後髮育，MyoD也從頭部往尾部錶達。在體節期，MyoD在中後部錶達。尾芽期MyoD的錶達量達到最大值。然而，尾芽期過後，頭部MyoD的錶達量急劇減少，但尾部仍然有彊烈的信號。在血流期仍然能在鱖魚尾部檢測到MyoD的微弱信號。到孵齣期在鱖魚魚眼的後方和胸部腹側檢測到MyoD的彊信號，而身體其他地方檢測不到信號。仔魚期沒有檢測到MyoD錶達的信號。
생기조절인자시기육형성적관건공제인소。통과RT-PCR방법극륭득도궐어주요생기조절인자MyoD서렬。궐어MyoD기인cDNA장도위813 bp，편마270개안기산，추도적안기산서렬함유1개bHLH결구。경여기타동물MyoD안기산서렬비대발현불동동물MyoD적bHLH결구역보수성고。통과합성지고신표기적반의RNA탐침대궐어불동발육계단적배태진행정체배태원위잡교，결과발현：MyoD최조재기절조기피검측도신호。반수착궐어체절종두부왕후발육，MyoD야종두부왕미부표체。재체절기，MyoD재중후부표체。미아기MyoD적표체량체도최대치。연이，미아기과후，두부MyoD적표체량급극감소，단미부잉연유강렬적신호。재혈류기잉연능재궐어미부검측도MyoD적미약신호。도부출기재궐어어안적후방화흉부복측검측도MyoD적강신호，이신체기타지방검측불도신호。자어기몰유검측도MyoD표체적신호。
Myogenic regulatoryfactors ( MRFs ) plays a key role in controlling myogenesis .The cDNA of MyoD in Siniperca chuatsi was cloned using RT-PCR.The sequence length of MyoD is 813 bp and encoding 270 amino acids.The MyoD includes a bHLH structures , which showed high conservation with other animals .The application of Dig labeled RNA probe on the detection of MyoD expression at different embryonic developmental sta-ges by whole-mount in situ hybridization .The results showed that MyoD expression was first detected in early myo-mere stage.As somite formed from rostral to caudal , the MyoD expressed from rostral to caudal .The most rich ex-pression stage was the caudal bud stage .After the caudal bud stage , the expression of the gene decreased rapidly , especially in the anterior somites where somatic cells were differentiated , but the posterior somites still showed a strong expression in the caudal somites .Blood circulation stage showed a weak expression .After hatching about 12 hours, there was a strong expression in the back of the eyes and the chest of the fish .At larval stage, no expres-sional signal was detected .
