Nishioka R S, de Jesus E G, Hyodo S
Department of Integrative Biology and Cancer Research Laboratory, University of California, Berkeley 94720.
Gen Comp Endocrinol. 1993 Jan;89(1):72-81. doi: 10.1006/gcen.1993.1010.
Oligonucleotide probes were synthesized for the mRNAs of a pair of tilapia prolactins (tPRL177 and tPRL188) and growth hormone (tGH) based on cDNAs for the hormones of Oreochromis niloticus and amino acid sequences for the hormones of O. mossambicus. The three 45mer probes were labeled with 35S for hybridization studies on pituitary sections of O. mossambicus adapted to fresh water (FW) or seawater (SW). Expression of tPRL mRNA in the rostral pars distalis was clearly evident with either PRL probe in adjacent sections in PRL cells of the rostral pars distalis; mRNAs of both PRLs were colocalized in the same cells. In addition, the tGH probe demonstrated expression of tGH mRNA specifically in GH cells in the proximal pars distalis. The hybridization signals for both PRLs were significantly greater in the rostral pars distalis of FW fish than in that of SW fish, as judged by computer-aided analysis. In addition, grain concentration for both PRLs was significantly greater over centrally located PRL cells of FW fish. In addition, although overall grain concentrations were lower in SW fish, there were significantly more grains over the centrally located PRL cells with the tPRL177 probe, whereas there was no difference with the tPRL188 probe. There was no detectable difference in the occurrence of tGH mRNA between FW and SW fish.
基于尼罗罗非鱼激素的cDNA和莫桑比克罗非鱼激素的氨基酸序列,合成了一对罗非鱼催乳素(tPRL177和tPRL188)和生长激素(tGH)mRNA的寡核苷酸探针。这三种45聚体探针用35S标记,用于对适应淡水(FW)或海水(SW)的莫桑比克罗非鱼垂体切片进行杂交研究。在相邻切片中,用任一PRL探针在远侧部吻端的PRL细胞中都能清楚地观察到tPRL mRNA的表达;两种PRL的mRNA共定位在同一细胞中。此外,tGH探针显示tGH mRNA特异性地在远侧部近端的GH细胞中表达。通过计算机辅助分析判断,FW鱼远侧部吻端的两种PRL的杂交信号明显大于SW鱼。此外,FW鱼位于中央的PRL细胞上两种PRL的颗粒浓度明显更高。此外,虽然SW鱼中的总体颗粒浓度较低,但用tPRL177探针时,位于中央的PRL细胞上的颗粒明显更多,而用tPRL188探针时则没有差异。FW鱼和SW鱼之间tGH mRNA的出现没有可检测到的差异。