Laser Capture Microdissection을 이용한 유전자 발현 연구(II) : 원시난포와 1차난포 유전자 발현의 차이에 대한 분석
성장을 멈추고 있는 원시난포(primordial follicle)에서 난포발달이 개시되어 1차난포(primary follicle)로 발달하는 조절기전은 잘 알려져 있지 않다. 이 초기 난포발달 과정에 관여하는 유전자를 알아내기 위해 suppression subtractive hybridization(SSH)을 사용하였다. 생후 1일과 5일째의 생쥐 난소로부터 얻은 cDNA로 forward와 reverse subtraction을 수행하여 각각 day1과
The present study was conducted to elucidate genes involved in the primordial-primary follicular transition. By using suppression subtractive hybridization, day1- and day5-subtracted cDNA libraries were obtained with the forward and reverse subtraction method, respectively. In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 genes with known functions were different between day1 and day5 ovaries. Four genes, GDF8, lats2, septin2, and wee1, from the day1 subtracted cDNA library, and 6 genes, HSP84, laminin2, MATER, MTi7, PTP, and wrn, from day5-subtracted cDNA library were chosen, and their differential expression was evaluated using RNAs from whole ovaries as well as captured primordial and primary follicles by laser captured microdissection. Results from the present study would provide insight for the future study on the mechanisms involved in primordial-primary follicle transition in the mouse in addition to the human ovary.