Thyroid hormones modulate the hypothalamo–hypophyseal–gonadal axis in teleosts: Molecular insights

Reproduction in fishes is influenced by thyroid hormones at various levels of gonadal cell differentiation and steroidogenesis. Thyroid hormones have recently also emerged as an important modulator of season- and photoperiod-dependent variations in the reproductive cycle with a possible effect on the hypothalamo–hypophyseal axis and pineal interactions. This review describes the current status of thyroid hormone research in relation to reproduction, with special emphasis on contributions to this field by Indian researchers including our laboratory. Evidence is provided for the multifocal action of thyroid hormones at various levels of the hypothalamo–hypophyseal–gonadal axis affecting reproduction. The underlying physiological and molecular mechanisms pertaining to thyroid hormone modulation of reproduction, such as gonadotropin-releasing hormone (GnRH) synthesis and release, androgen and gonadotropin receptor expression, gonadotropin (GTH) expression, and tissue sensitivity to GTHs are highlighted with relevant discussions of the current technical limitations, applications, and future perspectives of research in this field.     

巢式PCR法鉴定猪早期胚胎性别的研究

根据猪SRY基因序列设计合成2对巢式PCR引物作为性别鉴定引物,根据猪β-珠蛋白基因序列设计了一对引物作为内标引物建立了猪胚胎性别鉴定的PCR反应体系。公猪可以扩增出187bp的SRY基因片段和255bp的β-珠蛋白基因片段,母猪只能扩增出255bp的β-珠蛋白基因片段。由于巢式PCR只需3～8个细胞就可以在紫外灯下看到扩增结果,而常规PCR则需要较多的细胞,所以胚胎性别鉴定时使用巢式PCR效果更好。     

Sex change in coral reef fish

Gonadal differentiation can take many forms in fish, ranging from gonochorism, where individuals directly develop as male or female and finally possess only testis or ovaries at sexual maturation, to hermaphroditism where the same individuals can produce mature male and female gametes at some time in their lives. Hermaphrodite fish are, thus, an excellent model for studying the plasticity of sex determination and differentiation in vertebrates. We have shown that sex steroids play a principal role in sex differentiation and sex change in fish. Our laboratory implements several fish models that undergo sex change from female to male or male to female or in both directions. In this review, we will briefly discuss recent advances in our understanding of the mechanism of sex change in coral reef fish.     

Changes in thyroid hormone levels in eggs and larvae and in iodide uptake by eggs of coho and chinook salmon,Oncorhynchus kisutsch andO. tschawytscha

Developmental profiles of thyroxin (T₄), triiodothyronine (T₃) and radioactive iodide uptake were established for eggs and T₄ and T₃ profiles were established for larvae (whole-body, yolk-only and body-only) of coho and chinook salmon. T₄ and T₃ were consistently present in all samples. In eggs, hormone levels remained fairly constant in all cohorst for at least the first three weeks of incubation, but then fluctuated in both directions in some sample groups. Large increases in T₄ (from 9 ng/g to 245 ng/g) were seen in 1985 chinook eggs 28 days after fertilization. Radioactive iodide uptake (which was used as a possible indicator of thyroxinogenesis) increased at least 10-fold in both 1986 coho and chinook eggs from 23–30 days after fertilization. T₄ (62 ng/g) and T₃ (393 ng/g) were found in the bodies of 28-day-old 1986 chinook embryos. In whole larvae, hormone levels varied depending upon the cohort studied. In general, initial body-only concentrations of both T₄ and T₃ decreased as body weight increased, but before yolksac resorption was completed, both thyroid hormone content and concentration increased (except for chinook T₃). T₄ and T₃ content in larval yolk stayed constant as yolksac size decreased, resulting in increased thyroid hormone concentration in the yolksac. All of these data suggest that the initial source of thyroid hormones in coho and chinook salmon eggs is maternal, but that by approximately 3–4 weeks after fertilization, the developing embryos begin to produce their own thyroid hormones. After hatching, increases in tissue T₄ and T₃ concentration coupled with constant T₄ and T₃ content in diminishing yolksacs suggest that larvae also produce their own thyroid hormones; yolksac content then may reflect both the original maternal hormones and the larva-producted hormones.     

圆斑星鲽sox9基因的克隆与表达

本研究筛选到圆斑星鲽(Verasper variegatus)的性别相关基因sox9,并通过RACE技术获得了全长序列,基因全长为3287 bp,包括1431 bp的ORF,编码477个氨基酸,368 bp的5¢ UTR和1488 bp的3¢ UTR。在3¢ UTR中有多聚腺苷酸尾和加尾信号AATAAA。通过荧光定量PCR测定了sox9基因在圆斑星鲽成鱼不同组织中的表达水平,发现sox9基因在圆斑星鲽的脑、眼、鳃、心、肝、胆、肠、精巢、卵巢、肾和肌肉等各个组织中都有不同程度的表达。在鳃、脑和精巢组织中检测到较高水平的sox9转录,其中精巢中的转录水平显著高于其他组织,sox9基因在性腺中的表达显示出性别两相性差异。其在精巢中的表达水平要显著高于卵巢,说明sox9基因与雄性性腺发育相关。通过测定sox9基因在圆斑星鲽幼鱼不同发育时期(20、30、40、50、60、70和80日龄)的表达水平,发现其在20~50日龄表达量逐渐下降,在60日龄时表达量上升,推测表达量上升可能与幼鱼性腺分化相关。     

干湿交替灌溉和施氮量对水稻内源激素及氮素利用的影响

为探讨干湿交替灌溉与施氮水平对水稻根系内源激素水平及氮肥利用的影响,以连粳7号为材料,采用防雨棚土培试验,研究3个灌溉方式:浅水层灌溉、轻度干湿交替灌溉、重度干湿交替灌溉与3个氮肥水平(0、240和360 kg/hm2)对水稻根系内源激素(玉米素及玉米素核苷(Z+ZR)、生长素(IAA)、脱落酸(ABA))、叶片氮代谢酶活性(硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT))、植株氮素累积量及氮肥利用效率的影响及其耦合效应。研究结果表明:在相同施氮水平下,轻度干湿交替灌溉促进根系Z+ZR、IAA合成,提高叶片中NR、GS及GOGAT活性,氮肥吸收利用率显著提高(P0.05);重度干湿交替灌溉则抑制根系Z+ZR、IAA合成,降低叶片NR、GS及GOGAT活性,植株氮素累积量及氮肥利用效率显著降低(P0.05),而根系ABA含量则明显增加(P0.05);在相同灌溉方式下,根系Z+ZR、IAA含量、叶片氮代谢酶活性及氮肥累积量在保持水层及轻度干湿交替下随着施氮量的增加而增加,而在重度干湿交替灌溉下则随着施氮量的增加先增加后降低,中氮处理明显提高氮肥利用效率(P0.05)。相关分析表明:根系合成的Z+ZR、IAA及叶片中氮代谢酶活性与氮肥吸收利用率呈显著(P0.05)或极显著(P0.01)的正相关关系,而脱落酸含量则与氮肥吸收利用率呈极显著的负相关关系(P0.01)。根系合成的Z+ZR、IAA及叶片中氮代谢酶供氮效应为正效应,抽穗后,轻度干湿交替灌溉供水效应及耦合效应为正效应,而重度干湿交替灌溉则为负效应。该研究对探索水氮耦合机理,为水稻高产高效栽培实践提供理论及科学依据。     

沙棘微繁育苗产业化试验研究——水培沙棘茎段插穗不定根发生的机理

本文用水培方法研究了沙棘短茎段插穗生根的机理。结果表明,在用外源激素处理、控温控湿条件下,沙棘可以用极短的茎段(1～3芽段)插穗产生不定根,进而形成新的植物。沙棘插穗不定根形成有两种方式:一是茎段切面先产生愈伤组织,然后再分化为不定根;二是直接由茎皮层内分生组织产生不定根;且不定根产生具"茎多部位生根效应"。沙棘插穗不定根形成属于"温度敏感型",最适温度为16℃～26℃。外源激素对沙棘短茎段插穗生根表现出良好的诱导效应,而且对插穗苗的生长发育一定的影响,ABT优于GGR;"壮根素"效果优于生根粉,不仅促进发根,而且促进成熟不定根的早期结瘤。     

矮秆基因Rht_NM9在小麦株高建成中对内源激素含量的影响

普通小麦品种"南农9918"经甲基磺酸乙酯(ethyl methanesulfonate,EMS)诱变获得一个矮秆、多蘖、长穗突变体"NM9",在该突变体中定位到一个新的矮秆突变基因Rht_NM9。内源激素在普通小麦株高建成的过程中发挥重要的调节作用,为了解析Rht_NM9致矮的生理机制,本研究以南农9918及其矮秆突变体NM9为材料,利用酶联免疫吸附分析法(enzyme-linked immunosorbent assays,ELISA)测定了不同生育期各节间内源赤霉素(GA)、生长素(IAA)、脱落酸(ABA)和玉米素核苷(ZR)的含量,分析小麦发育关键时期的内源激素含量变化与株高的关系。结果表明,在孕穗期和抽穗期,矮秆突变体NM9中GA、ABA含量均显著高于南农9918,而ZR含量则显著低于南农9918,IAA含量在南农9918和突变体NM9之间无明显差异。此外,突变体NM9各节间中GA/ABA比值显著高于南农9918,而IAA/ABA、(IAA+GA)/ABA、ZR/ABA比值显著低于南农9918。以上结果表明小麦株高受多种激素调控,突变体中内源ABA含量升高,IAA/ABA和ZR/ABA比值降低会抑制植物株高伸长。     

糜子愈伤诱导及再分化最适条件研究

研究旨在应用组织培养技术探究愈伤诱导及再分化的最适条件,以期为糜子建立遗传转化体系奠定基础。采用5个糜子品种研究愈伤诱导及再分化的最适条件。利用植物激素2,4-二氯苯氧乙酸(2,4-D)诱导茎尖愈伤组织,筛选出发芽率高、愈伤组织诱导率高、染菌率低的品种,并对诱导愈伤最适激素浓度及配比进行了鉴定。结果显示,‘冀黍2号’出芽率高、愈伤组织诱导率高、染菌率低,适宜进行组织培养;2,4-D对‘冀黍2号’愈伤诱导效果最好,最适浓度为2.5 mg/L,诱导率达86.67%,淡黄色块状,结构紧密,质地较硬,继代培养后形成的胚性愈伤组织状态更好。再分化过程中,2.5 mg/L 2,4-D+3.5 mg/L TDZ时,出现明显的嫩芽。该试验获得‘冀黍2号’愈伤诱导及再分化的最适条件,可为糜子再生体系的构建提供参考。     

外源调节物质对盐胁迫下植物生长调控研究进展

为了探究外源物质对植物耐盐性的调控机理及进一步利用外源调节物质提高作物耐盐性,归纳了五大类传统植物激素（生长素、赤霉素、乙烯、脱落酸、细胞分裂素）以及褪黑素、水杨酸、多胺、油菜素类固醇、茉莉酸类等外源生长调节物质对盐胁迫下植物生长的调控情况。同时总结了硅、钙等离子类外源调节物对盐胁迫下植物生长的调节作用,多种外源调节物质可通过增强光合作用、提高渗透势、增加抗氧化酶活性及减少离子毒害等方式来减轻盐害。本文为进一步利用单一或复合外源调节物质来缓解作物盐害盐提供理论依据。