Fundamental studies of a developing heteromorphic sex chromosome pair in hynobiid salamanders.

小蝾螈发育中的异形性染色体对的基础研究。

• 批准号: 11833016
• 负责人: KOHNO Sei-ichi
• 金额: $ 2.3万
• 依托单位: TOHO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11833016/
• 关键词: sex chromosome; Hynobius; heteromorphism; repetitive DNA; microdissection; Southern blot hybridization; W chromosome; PCR; 異型性; 反復性DNA; チョウセンサンショウウオ

Hynobius leechii in Cheju Island (Korea) has a pair of female heteromorphic sex chromosomes in which the W chromosome shows polymorphism in its size and shape according to its habitat (Ikebe unpublished data). On the other hand, Hynobius tokyoensis in Kanto district (Japan) also has the same kind of sex chromosomes with polymorphism and the W chromosomes in this species are different from those of H.leechii. Heteromorphism in the two species are clearly caused by increase of heterochromatin in their W chromosomes. On the contrary, most of H.neburosus which inhabits between the habitats of H.leechii and H.tokyoensis, does not have a heteromorphic sex chromosome pair (Ikebe ＆ Kohno 1979, Ikebe et al. l987). Namely, it is natural to understand that the heteromorphic sex chromosome pairs should be developed independently in these two species.For obtaining the fundamental knowledge in promoting factors of heteromorphism in the sex chromosome pair of the two species, first, we tried to detect the repeat sequences which amplified in W chromosomes in H.leechii. Microdissection of the chromosomes was carried on the heterochromatin region and the centromeric region of the W chromosomes in metaphases of embryos from Songpanak (collected in 1998) and Kyorairi (collected in 2000). By using the microcloning, the three step degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR), and the Southern blot hybridization, one DNA fragment which is originated in genomic DNA of H.leechii was isolated. The fragment was seqenced and the size was 208 base pairs. It was not specific to the female genomic DNA of H.leechii..In near future, we have to isolate the sequence which consists of heterochromatin in W chromosomes of H.leechii and have to compare it to that of H.tokyoensis.

韩国济州岛的李氏小鲵具有一对雌性异形性染色体，其中W染色体根据其栖息地在大小和形状上表现出多态性（Ikebe未发表的数据）。另一方面，日本关东地区的小鲵（Hynobius tokyoensis）也具有相同类型的性染色体，并具有多态性，其W染色体与李氏小鲵（Hynobius leechii）不同。这两个物种的异形现象明显是由于它们的W染色体上异染色质的增加造成的。相反，大多数栖息在H.leechii和H.tokyoensis栖息地之间的H. neberosus没有异形性染色体对（Ikebe & Kohno 1979，Ikebe et al. 1987）。为了获得这两个物种的性染色体对中异形性的促进因素的基础知识，我们首先检测了在利氏血蜱W染色体上扩增的重复序列。对松潘（1998年采集）和京来（2000年采集）胚胎中期W染色体的异染色质区和着丝粒区进行了染色体显微切割。通过微克隆、三步简并寡核苷酸引物聚合酶链反应（DOP-PCR）和Southern印迹杂交，从利氏血吸虫基因组DNA中分离到一个DNA片段。对该片段进行测序，其大小为208个碱基对。这是不是特定的女性基因组DNA的H. leechii。在不久的将来，我们必须分离出的异染色质的序列在H.leechii的W染色体上，并将其与H. tokyoensis进行比较。

项目成果

石田かおり: "性染色体の同定並びに形態分化に関する基礎的研究"東邦大学大学院理学研究科. 95 (2000)

石田香织：“性染色体鉴定和形态分化的基础研究”东邦大学理学研究生院95（2000）。

Ikebe,C.: "Cytogenetic Studies of Hynobiidae (Urodela). XXII. Banded karyotypes and sex chromosomes in Hynobius tenuis Nambu."Chromosome Sci.. 4・(in press). (2000)

Ikebe, C.：“Hynobiidae (Urodela) 的细胞遗传学研究。XXII。Hynobius tenuis Nambu 的带状核型和性染色体。”染色体科学.. 4·（印刷中）。

Ikebe,C.: "Cytogenetic studies of Hynobiidae (Urodela).XU1.Comparative C-banded karyotype analysis of Pseudohynobius flabomaculatus (Fei et Ye), Ranodon shihi (Liu) and Batrachuperus pinchonii(David)."Chromosome Res.. 8. 265-272 (2000)

Ikebe,C.：“Hynobiidae (Urodela) 的细胞遗传学研究。XU1。Pseudohynobius flabomaculatus (Fei et Ye)、Ranodon shihi (Liu) 和 Batrachuperuspinkonii(David) 的比较 C 带核型分析。”染色体研究 8。

石田かおり: "性染色体の同定並びに形態変化に関する基礎的研究"東邦大学理学研究科. 95 (2000)

石田香织：“性染色体的鉴定和形态变化的基础研究”东邦大学理学研究生院95（2000）。

Ikebe,C.: "Pairing behavior of sex lampbrush chromosomes in Hynobius leechii"Chromosome Sci.. 3. 152-152 (1999)

Ikebe,C.：“Hynobius leechii 性灯刷染色体的配对行为”《染色体科学》3. 152-152 (1999)