CAJ | 학술논문

鸣禽的发声行为及其前脑发声控制核团存在显著的性双态性,这种性双态性是在发育过程中逐步建立起来的.鸣禽白腰纹鸟35日龄(P35)前,雌、雄鸟前脑发声控制神经核RA(Robust nucleus of arcopallium) 不存在性别差异;到45日龄(P45)时,雌、雄鸟RA体积已经出现了显著的差异.推测这种神经结构的性双态性的形成与各种基因的不同时空表达有关.本实验以发育不同阶段(P35时的雌性, P45时的雌性和雄性)的雌、雄白腰纹鸟RA核团为研究对象,用mRNA差异显示银染技术来寻找前脑上述三组材料间的差异表达基因,试图发现调控RA核团性双态性发育的关键基因.本实验共选用了4种锚定引物与16种随机引物所组成的64组引物进行DDRT-PCR(mRNA differential display and reverse transcription-polymerase chain reaction)反应.第一次扩增后,在测序胶上显示出1 158条cDNA扩增产物,其中273个为差异表达.长度分布于100-500 bp之间的差异条带共161条.本实验重点研究了P35雌性和P45雄性共表达或P45雌性单独表达的条带(共19条),推测这些基因片段可能与引起RA核团中的细胞凋亡和性双态性的形成有关.二次PCR扩增后,进一步筛选出16个差异条带,其中11个为P35雌性和P45雄性共同表达的,5个是P45雌性特异表达的.本实验对其中一个差异片段T3进行了测序和分析,发现该片断对应的氨基酸序列与鸡(Gallus gallus)Bcl复合体有81%的同源性,由此推测Bcl-2基因家族有可能在促进RA核团的细胞凋亡中发挥作用,但有关T3片段的具体功能还需进一步深入研究 [动物学报 51(1):149-155,2005].
명금적발성행위급기전뇌발성공제핵단존재현저적성쌍태성,저충성쌍태성시재발육과정중축보건립기래적.명금백요문조35일령(P35)전,자、웅조전뇌발성공제신경핵RA(Robust nucleus of arcopallium) 불존재성별차이;도45일령(P45)시,자、웅조RA체적이경출현료현저적차이.추측저충신경결구적성쌍태성적형성여각충기인적불동시공표체유관.본실험이발육불동계단(P35시적자성, P45시적자성화웅성)적자、웅백요문조RA핵단위연구대상,용mRNA차이현시은염기술래심조전뇌상술삼조재료간적차이표체기인,시도발현조공RA핵단성쌍태성발육적관건기인.본실험공선용료4충묘정인물여16충수궤인물소조성적64조인물진행DDRT-PCR(mRNA differential display and reverse transcription-polymerase chain reaction)반응.제일차확증후,재측서효상현시출1 158조cDNA확증산물,기중273개위차이표체.장도분포우100-500 bp지간적차이조대공161조.본실험중점연구료P35자성화P45웅성공표체혹P45자성단독표체적조대(공19조),추측저사기인편단가능여인기RA핵단중적세포조망화성쌍태성적형성유관.이차PCR확증후,진일보사선출16개차이조대,기중11개위P35자성화P45웅성공동표체적,5개시P45자성특이표체적.본실험대기중일개차이편단T3진행료측서화분석,발현해편단대응적안기산서렬여계(Gallus gallus)Bcl복합체유81%적동원성,유차추측Bcl-2기인가족유가능재촉진RA핵단적세포조망중발휘작용,단유관T3편단적구체공능환수진일보심입연구 [동물학보 51(1):149-155,2005].
Song behavior and its song-control nuclei (such as the robust nucleus of the arcopallium-RA) are highly sexually dimorphic in passerine songbirds. These dimorphisms are not present early in development, but arise through processes during sensitive periods in life. There are no sex differences in RA volume and neuron numbers of white-rumped munia at P35 (date after birth), but sexual dimorphism in RA volume became very markedly at P45. It is thought that the development of neural dimorphism is controlled by genes. In this experiment, the female and male RAs at developmental ages P35 (females only) and P45 (females and males) were studied by mRNA differential display (DD) technology. For the DDRT-PCR, combinations of the four T12MN primers together with sixteen arbitrary primers, AP1-16, were used. This analysis yielded 1 158 bands in total, of which 273 bands were differentially generated. There were 161 differential bands which size was between 100 bp and 500 bp. The aim of this study was focused on the cDNAs co-expressed in P35 female and P45 male (sixty-one bands) or expressed only in female at P45 (nineteen bands), which might play a role in the dimorphic development of RA. After the second PCR amplification, sixteen differential bands were screened out, of which eleven were co-expressed in P35 female and P45 male, five were only expressed in female at P45, which might underlie the genetic mechanism of the RA dimorphic development in white-rumped munia. A differential segment named T3, which had 81% homology with Bcl complex in chichen, might be a member of Bcl-2 protein family or relate to the retaining of RA volume, was chosen to sequence and analyse. However, the information about T3 segment in detail needed further research. These results show that mRNA DD methodology can represent a potential tool for investigation of the gene expression in the brain of songbird development [Acta Zoologica Sinica 51(1):149-155,2005].