中国林蛙蝌蚪变态过程中甲状腺的发育

两栖动物幼体变态过程是受甲状腺激素所精密调控的。对中国林蛙Rana chensinensis变态前期、临近变态期、变态高峰期和变态完成期的蝌蚪进行了外部形态指标(全长、体长、尾长和后肢长等)的测定,采用解剖和组织学方法对其甲状腺进行了组织学观察,并对蝌蚪外部形态指标与甲状腺形态机能之间的相关性进行了统计分析。结果显示,中国林蛙蝌蚪甲状腺分泌甲状腺激素机能活性的峰值出现于变态高峰期的前肢伸出期。统计分析表明,蝌蚪外部形态指标全长和后肢长的发育信息也可以反映其甲状腺分泌甲状腺激素的机能活性。     

水位变化影响蝌蚪存活、形态表型和植物血凝素反应

为探究黑斑侧褶蛙蝌蚪形态和生理特征对水位变化的响应,对480只大小相似的蝌蚪进行快速、中速和慢速干涸处理,统计各发育时期时长和存活率,测定变态完成后幼蛙的身体和内脏器官大小,以及对植物血凝素(PHA-P)的反应。结果表明: 对照组蝌蚪的变态时长最长,显著长于快速、中速和慢速干涸组;不同处理变态存活率在72.5%～90.8%,其中对照组最高,快速干涸组最低。对照和慢速干涸组幼蛙的体重和体长均显著高于中速和快速干涸组,快速干涸组体宽、体重与体长的比值、胴体湿重以及肺和脂肪体的湿重系数均最低,但心、脾、肾和消化器官的湿重系数和消化道各段的长度系数均无明显的组间差异。不同处理幼蛙对PHA-P的最大反应值均出现于注射后3 h,且中速和慢速干涸组的最大反应值均显著高于对照组,而快速干涸组与各组均无显著差异。表明黑斑侧褶蛙蝌蚪可通过加速变态发育进程来应对干旱胁迫,但会出现个体变小、细胞介导的免疫能力下降,不利于其成功登陆。     

外来捕食者克氏原螯虾对黑斑侧褶蛙不同发育阶段幼体表型与运动能力的影响

外来物种入侵是造成本土无尾两栖类种群下降和灭绝的最严重威胁之一。在中国南部丽水地区,克氏原螯虾(Procambarus clarkiion)是一种捕食本土无尾类蝌蚪的入侵物种。在实验室条件下,通过对黑斑侧褶蛙(Pelophylax nigromaculatus)受精卵和蝌蚪先后进行克氏原螯虾捕食风险的不同处理,检测孵出蝌蚪的形态特征、中期蝌蚪的形态特征、游泳表现及活动水平、变态时间与大小以及幼蛙跳跃表现,评估克氏原螯虾的捕食风险对黑斑侧褶蛙不同生长阶段幼体生长发育、运动能力及行为的综合影响。单因素方差分析表明,捕食者长期存在条件下黑斑侧褶蛙孵出蝌蚪的体长、尾长和尾肌宽均显著短于无捕食者的空白对照组和捕食者仅存在2 d的实验组,后两者处理下的蝌蚪体长差异显著,但尾长和尾肌宽差异不显著。长期捕食者存在条件下黑斑侧褶蛙蝌蚪的游泳总路程和平均游速均显著小于无捕食者和捕食者存在2 d的实验组,而蝌蚪体长、尾长、尾鳍高和尾肌高的组间差异均不显著。有捕食者存在下黑斑侧褶幼蛙的体长和体重均显著大于无捕食者和捕食者存在2 d的实验组的情况,而变态时间和幼蛙跳跃距离组间差异不显著。两因素方差分析表明,测试时间、捕食者设置和两因素的相互作用均显著影响蝌蚪的活动频次。不同实验时间,长期捕食者存在实验组蝌蚪活动频次均小于无捕食者和捕食者存在2 d的实验组。无捕食者实验组蝌蚪的活动频次前期高于捕食者存在2 d的实验组,后期低于捕食者存在2 d的实验组。线性回归分析结果显示,捕食者存在2 d实验组蝌蚪相对活动频次与检测时间相关不显著,长期捕食者存在实验组相对蝌蚪活动频次与检测时间呈显著的正相关。本研究表明了黑斑侧褶蛙蝌蚪能够感知来自入侵捕食者克氏原螯虾的捕食风险,诱导产生较小的孵出个体、蝌蚪较低的活动水平,但这种不足可通过后期补偿生长,不影响进入陆地生境的变态个体的表型与运动能力。     

镉对黑斑蛙蝌蚪生长发育的影响

两栖动物幼体的生长发育过程对环境变化和化学污染物都很敏感,为研究镉对两栖动物幼体生长发育的影响,将37—38期黑王证蛙（Rana．nigromaculata）蝌蚪暴露于含不同浓度镉的水中饲养,直至完全变态,观察幼体生长发育过程,记录蝌蚪完全变态所需时间,平均体重和平均长度增长率的测定。结果表明,随着Cd^2＋浓度的升高,蝌蚪的啤均体重和平均长度的增长率降低,完全变态所需时间增长。说明Cd^2＋污染可抑制黑斑蛙蝌蚪的生长发育,延缓变态。     

黑斑侧褶蛙蝌蚪断尾后的补偿生长和发育研究

动物在经历不利的生长条件或环境后往往出现补偿生长。研究了黑斑侧褶蛙(Pelophylax nigromaculatus)蝌蚪经历来自食蚊鱼(Gambusia affinis)捕食造成断尾损伤后的补偿生长模式、断尾损失对蝌蚪游泳能力以及变态时间与大小的影响。结果表明,经历捕食压力后,全尾组和1/3断尾组蝌蚪的体长显著大于1/2断尾组蝌蚪的体长,全尾组和1/3断尾组之间的蝌蚪体长差异不显著;第19天时,1/2断尾组蝌蚪经过补偿生长后体长显著大于全尾组蝌蚪,1/3断尾组与全尾组和1/2断尾组之间的蝌蚪体长差异均不显著;三个实验组之间蝌蚪尾长差异不显著;全尾组蝌蚪的疾游速显著大于1/2断尾组蝌蚪的疾游速,1/3断尾组蝌蚪的疾游速与全尾组和1/2断尾组之间差异不显著,表明严重断尾对蝌蚪疾游速产生了消极影响。三个实验组蝌蚪的变态时间和变态前后形态差异均不显著。黑斑侧褶蛙蝌蚪能够在变态前调整生长轨迹补偿早期捕食风险造成的生长损失,断尾损失并不影响变态时间与大小,断尾超过一半的蝌蚪经过补偿生长后仍要付出一定的运动代价。     

敌敌畏对多疣狭口蛙不同发育期蝌蚪的急性毒性影响

为了解农药对不同发育期蝌蚪的毒性,以多疣狭口蛙Kaloula verrucosa蝌蚪为对象,采用水生生物急性毒性试验方法,研究杀虫剂敌敌畏(DDVP)对25期、26期、35期和42期蝌蚪的急性毒性影响.结果表明,在水温19.5～22.0℃条件下,敌敌畏对各期蝌蚪96 h LC50分别为1.13、1.43、1.38和0.61 mg/L,各期蝌蚪对敌敌畏的敏感顺序为42期>25期>35期>26期.由此得出敌敌畏对25期、26期和35期蝌蚪属中等毒性,对42期蝌蚪则表现为高毒.因此建议农药施用应尽量避开蝌蚪的变态期进行.蛙类胚胎期(出膜后)随着发育对外界环境的耐受性增强,蝌蚪期随着发育对外界环境的敏感性增强.鉴于42期蝌蚪意外死亡率高的特点,认为不宜采用变态期蝌蚪进行急性毒性试验.     

中国林蛙蝌蚪的口器发育

采用扫描电镜和组织学技术观察了中国林蛙(Rana chensinensis)蝌蚪发育过程中口器外部形态结构的变化,以及中国林蛙蝌蚪口器内部结构特征.结果表明,在口器发育的初期,角质颌最先出现,接着出现唇乳突以及唇齿;在变态高峰期(G4l～G42),口器结构如唇齿、角质颌和唇乳突则是按以下顺序消失的,即唇齿最先消失,其次...     

水位对黑斑侧褶蛙蝌蚪形态表型、 白细胞数量和应激反应的影响

为探究水位变化对黑斑侧褶蛙（Pelophylaxnigromaculatus）蝌蚪形态表型、免疫功能和应激反应能力的影响,实验室内测定了水位恒定组以及水位快速、中速和慢速下降组37期蝌蚪发育至变态完成期的变态时长、身体大小和血液中各型白细胞的百分比,以及嗜中性粒细胞与淋巴细胞比值的变化。结果表明,水位快速下降显著缩短了蝌蚪的变态时长（P <0.05）,体长下降明显（P <0.05）,体重和体宽有所降低,但组间差异不显著（P> 0.05）。嗜中性粒细胞的百分比快速下降组显著高于其他处理组,单核细胞的百分比水位恒定组显著高于中速下降组（P <0.05）,嗜酸性粒细胞、嗜碱性粒细胞和淋巴细胞的百分比均无明显的组间差异（P>0.05）;嗜中性粒细胞与淋巴细胞比值快速下降组显著高于其他处理组（P <0.05）。水位加速下降导致登陆后幼蛙的个体变小,由嗜中性粒细胞和单核细胞所代表的天然免疫能力变化趋势不同,应激反应能力增强,可能不利于幼蛙的存活。     

几种农药对黑斑蛙黑斑蛙胚胎及蝌蚪的毒性

本文报道了常用的几种农药对黑斑蛙胚胎及蝌蚪发育的毒性影响。结果表明 :农药对黑斑蛙胚胎、蝌蚪期的毒性大小依次为使它隆 >多效唑 >多菌灵 >异丙隆 >甲胺磷。胚胎期死亡时表现为胚体腐烂 ;蝌蚪期死亡时则表现为头部膨大 ,尾部缩小 ,弯曲。畸形在两个时期都表现为腹部膨大呈透明状和尾部弯曲     

蝌蚪加服甲状腺激素后对生长发育的影响

在学习了高中《生物》第４章第２节“甲状腺激素的作用”后,我带着生物课外小组的学生做了一个有趣的课外实验。１　实验目的观察甲状腺激素对蝌蚪发育过程中形态变化的影响。２　动物与器材同一雌蛙产下的受精卵培育成蝌蚪（１０ｍｍ左右）４０只,分成实验组和对照组。１０００ｍｌ广口瓶２个、有孔匙羹、平底玻璃碟１个、方格纸（１ｍｍ×１ｍｍ）１３张、绿藻、甲状腺制剂（把药用甲状腺片与等量面粉一起研磨碎,加水调成糊状,阴干后再研碎成粉末状即成）５ｇ。３　方法与步骤３．１　将蝌蚪分成实验组与对照组分别饲养于下列２个广口…     

Adenohypophysial-thyroid activity of the tiger salamander, Ambystoma tigrinum, as a function of metamorphosis and captivity

The aim of this study was to determine the timing of adenohypophysial activation during metamorphosis of the tiger salamander, Ambystoma tigrinum. It consisted of two parts: 1) determination of plasma thyroid hormone concentrations and analysis of thyroid gland histology as a function of metamorphic stage and 2) analysis of the time-course of uptake of 125I by the thyroids during metamorphosis as an indicator of endogenous thyrotropin (TSH) levels. Significant increases in both triiodothyronine (T3) and thyroxine (T4) first were evident at the onset of metamorphic climax (stage II). Maximum levels of both hormones were not observed, however, until the completion of gill resorption (stage VII). No changes in thyroid histology were observed that could be unambiguously related to metamorphic transformation. The thyroids accumulated 125I in a slow but linear fashion in premetamorphic larvae (stage I). However, uptake exhibited a rapid peak during early climax (stage II), before maximum concentrations of thyroid hormones were observed. In addition, uptake was maintained above premetamorphic levels at stage VII, in conjunction with maximum levels of T4 and T3. Captivity alone produced a small but significant increase in plasma concentrations of T3. It produced no significant effect on either thyroid histology or uptake of 125I. These results indicate that adenohypophysial activation occurs rapidly and is maximal at the onset of metamorphic climax.     

ON THE DEVELOPMENT OF THE ULTIMOBRANCHIAL GLAND IN AN ANURAN,RANA JAPONICA JAPONICA*

The development of the ultimobranchial gland (UBG) was studied from its earliest emergence in Rana japonica japonica. UBG primordia appear at stage 22 as outfoldings of the pharyngeal epithelium of the 6th visceral pouch in both sides of the body. At stage 24, they separate from the pharyngeal wall, and then become follicular at stage 25. During stages 27–30, which are just prior to metamorphic climax, the UBG seems to be activated, as shown by secondary follicle formation and pseudostratification of the follicle epithelium. At and after the completion of metamorphosis, the UBG shows a histological profile indicating low activity.     

Ontogeny of the Thyroid Glands During Larval Development of South American Horned Frogs (Anura,Ceratophryidae)

The role of thyroid hormone (TH) in anuran metamorphosis has been documented from a variety of approaches, but the sequence of morpho-histological development of the thyroid glands that produce the secretion of the hormone was assumed invariant from studies of relatively few species even when the effects of environmental influences on larval development and metamorphosis have been largely documented. There are anurans in which developmental and growth rates diverge, and the resulting heterochrony in growth and development produces giant/miniature tadpoles, and or rapid/delayed metamorphosis suggesting changes of the activity of the thyroid glands during larval development. Herein, we analyze the morpho-histological variation of the thyroid glands in larval series of Ceratophrys cranwelli, Chacophrys pierottii, Lepidobatrachus laevis and L. llanensis that share breeding sites along semiarid environments of the Chaco in South America, belong to a monophyletic lineage, and present accelerated patterns in growth and development in order to have a morphological evidence about a possible shift of TH physiology. We describe gross morphology and histology of the thyroid glands and find features shared by all studied species such as the presence of supernumerary heterotopic follicles; changes in the volume and number of follicles towards the metamorphic climax, and cuboidal epithelia with occasional intra-cellular vacuoles as signs of low glandular activity without a manifest peak at the climax as it was assumed for anurans. We discuss different lines of evidence to interpret sources of extra supplement of TH to support the rapid metamorphosis. These interpretations highlight the necessity to design a research program to investigate the endocrine variation during development of ceratophryids taking in account their morphology, physiology and ecology in order to learn more about the effects of environmental and developmental interactions involved in the anuran evolution.     

Gene switching at Xenopus laevis metamorphosis

During the climax of amphibian metamorphosis many tadpole organs remodel. The different remodeling strategies are controlled by thyroid hormone (TH). The liver, skin, and tail fibroblasts shut off tadpole genes and activate frog genes in the same cell without DNA replication. We refer to this as “gene switching”. In contrast, the exocrine pancreas and the intestinal epithelium dedifferentiate to a progenitor state and then redifferentiate to the adult cell type. Tadpole and adult globin are not present in the same cell. Switching from red cells containing tadpole-specific globin to those with frog globin in the liver occurs at a progenitor cell stage of development and is preceded by DNA replication. Red cell switching is the only one of these remodeling strategies that resembles a stem cell mechanism.     

Histological changes in the pituitary-thyroid axis during spontaneous and artificially-induced metamorphosis of larvae of the flounder Paralichthys olivaceus

Summary Histological changes in the pituitary TSH cells and in the thyroid gland of flounder (Paralichthys olivaceus) larvae during spontaneous or artificially induced metamorphosis were studied. Activity of the immunoreactive TSH cells (IrTSH cells) gradually increased during premetamorphosis, reaching the highest level in prometamorphic larvae, and the cells were degranulated in metamorphic climax. The IrTSH cells were most inactive at the post-climax stage. The thyroid gland was morphologically the most active in metamorphic climax when the degranulation occurred in the pituitary IrTSH cells, and appeared inactive at post-climax. A few weeks after metamorphosis, both the IrTSH cells and the thyroid gland appeared to be activated again in the benthic, juvenile flounder. Administration of thyroxine or thiourea revealed negative feedback regulation of the pituitary-thyroid axis in flounder larvae. These results indicate that activation of the pituitary-thyroid axis induces metamorphosis in the flounder.     

Expression and regulation of miR-1, -133a, -206a, and MRFs by thyroid hormone during larval development in Paralichthys olivaceus

MiR-1, miR-133a, and miR-206a have been identified as muscle-specific miRNAs. They play multiple crucial roles in the regulation of muscle development. Here, we show that these miRNAs were differentially expressed during the larval development of flounder, and specifically expressed in skeletal muscle and heart in adult tissues/organs. The expression levels of these miRNAs were significantly changed by thyroid hormone (TH) or thiourea (TU) treatment during metamorphosis from 17 dph (days post hatching) to 42 dph. In addition, the expression levels of MyoD and Myf5 mRNAs markedly increased at 14 dph (pre-metamorphosis) compared to metamorphic stages, and their expression levels are far above the myogenin during larval development. Moreover, these MRFs (myogenic regulatory factors) expression were directly or indirectly regulated by thyroid hormone or thiourea during metamorphosis. All the results suggest that miRNAs and MRFs might be involved in signaling pathway of TH or TU-mediated flounder metamorphosis.     

甲状腺激素在两栖动物变态过程中的作用

两栖动物的幼体变态是研究甲状腺激素调节组织和器官重构的理想模式。本文主要综述了近年来两栖动物甲状腺激素合成过程中3种脱碘酶D1、D2和D3的特点及其生物学功能;甲状腺激素受体的蛋白结构、类型和机能;以及甲状腺激素对两栖动物幼体变态过程中各个类型组织和器官重构的调节;甲状腺激素、甲状腺激素受体和脱碘酶的互作,并展望了今后的研究方向。     

The effects of corticosteroid hormones and thyroid hormones on lymphocyte viability and proliferation during development and metamorphosis of Xenopus laevis

Abstract. Metamorphosis in the South African clawed frog, Xenopus laevis , is characterized by a striking loss of lymphocytes in the thymus, liver, and spleen. Changes in the proliferative responses of splenocytes and thymocytes to T cell mitogens and semi-allogeneic cells are also observed at metamorphosis. Because the levels of circulating thyroid hormones (TH) and corticosteroid hormones (CH) increase dramatically during the climax of metamorphosis, we have investigated the possible role of TH and CH as mediators of the changes in lymphocyte numbers or lymphocyte function. Here we report on the in vitro effects of CH and TH on lymphocyte viability and on phytohemagglutinin-P (PHA)-stimulated lymphocyte proliferation at prometamorphosis and climax of metamorphosis. We have observed consistently significant inhibition of proliferation by corticosterone. In contrast, we have observed inconsistent inhibition of proliferation by both thyroxine (T₄) and triiodothyronine (T₃). In short-term studies, the viability of thymocytes and splenocytes was reduced in the presence of CH but not TH.
These observations are consistent with a hypothesis that loss of larval lymphocytes and changes of lymphocyte function at metamorphosis may be due to elevated concentrations of CH rather than TH.
Because CH have been shown to enhance TH-induced effects during metamorphosis, we looked at the combined effects of these agents on PHA-stimulated lymphocyte proliferation. While each agent was inhibitory in several experiments, there was no significantly greater inhibition when splenic lymphocytes were cultured with both.     

One of the duplicated matrix metalloproteinase-9 genes is expressed in regressing tail during anuran metamorphosis

The drastic morphological changes of the tadpole are induced during the climax of anuran metamorphosis, when the concentration of endogenous thyroid hormone is maximal. The tadpole tail, which is twice as long as the body, shortens rapidly and disappears completely in several days. We isolated a cDNA clone, designated as Xl MMP-9TH, similar to the previously reported Xenopus laevis MMP-9 gene, and showed that their Xenopus tropicalis counterparts are located tandemly about 9 kb apart from each other in the genome. The Xenopus MMP-9TH gene was expressed in the regressing tail and gills and the remodeling intestine and central nervous system, and induced in thyroid hormone-treated tail-derived myoblastic cultured cells, while MMP-9 mRNA was detected in embryos. Three thyroid hormone response elements in the distal promoter and the first intron were involved in the upregulation of the Xl MMP-9TH gene by thyroid hormone in transient expression assays, and their relative positions are conserved between X. laevis and X. tropicalis promoters. These data strongly suggest that the MMP-9 gene was duplicated, and differentiated into two genes, one of which was specialized in a common ancestor of X. laevis and X. tropicalis to be expressed in degenerating and remodeling organs as a response to thyroid hormone during metamorphosis.     

Sequential up-regulation of thyroid hormone β receptor,ornithine transcarbamylase,and carbamyl phosphate synthetase mRNAs in the liver of Rana catesbeiana tadpoles during spontaneous and thyroid hormone-induced metamorphosis

During both spontaneous and thyroid hormone (TH)-induced metamorphosis, the Rana catesbeiana tadpole undergoes postembryonic developmental changes in its liver which are necessary for its transition from an ammonotelic larva to a ureotelic adult. Although this transition ultimately results from marked increases in the activities and/or de novo synthesis of the urea cycle enzymes, the precise molecular means by which TH exerts this tissue-specific response are presently unknown. Recent reports, using RNA from whole Xenopus laevis tadpole homogenates and indirect means of measuring TH receptor (TR) mRNAs, suggest a correlation between the up-regulation of TRβ-mRNAs and the general morphological changes occurring during amphibian metamorphosis. To assess whether or not this same relationship exists in a TH-responsive tissue, such as liver, we isolated and characterized a cDNA clone containing the complete nucleotide sequence for a R. catesbeiana urea cycle enzyme, ornithine transcarbamylase (OTC), as well as a genomic clone containing a portion of the hormone-binding domain of a R. catesbeiana TRβ gene. Through use of these homologous sequences and a heterologous cDNA fragment encoding rat carbamyl phosphate synthetase (CPS), we directly determined the relative levels of the TRβ, OTC, and CPS mRNAs in liver from spontaneous and TH-induced tadpoles. Our results establish that TH affects an up-regulation of mRNAs for its own receptor prior to up-regulating CPS and OTC mRNAs. Moreover, results with cultured tadpole liver demonstrate that TH, in the absence of any other hormonal influence, can affect an up-regulation of both the TRβ and OTC mRNAs. © 1992 Wiley-Liss, Inc.