处于性腺不同发育阶段斜带石斑鱼垂体的EST表达差异分析

斜带石斑鱼是重要的海产经济鱼类,在其个体发育过程中存在先雌后雄的天然性反转现象。垂体是调节生长和生殖等生理过程的重要内分泌器官。构建了斜带石斑鱼分别处于卵巢发育起始和性反转后期的垂体SMART cDNA的质粒文库,并通过测序分别筛选到232个和258个表达序列标签(expressed sequence tags, EST)。将所得EST与GenBank数据库中的序列进行比对,结果表明,处于卵巢发育起始和性反转后期斜带石斑鱼垂体EST中,激素所占比例均为最高,分别为40.5%和34.9%。进一步比较分析了这两个性腺发育时期斜带石斑鱼垂体EST中各种激素相对表达丰度,表明生长/催乳激素家族（GH、PRL和SL）和阿黑皮素原（POMC）表达水平下降;促性腺激素α亚基（GTHα）表达水平急剧上升,促滤泡激素β亚基（FSHβ）、促黄体激素β亚基（LHβ）表达水平上升。     

斜带石斑鱼PACAP的原核表达及活性分析

自1989年从绵羊下丘脑提取物发现垂体腺苷酸环化酶激活多肽（Pituitary adenylate cyclase activating polypeptide,PACAP）以来（Miyata et al．,1989）,已证明它能促进垂体激素释放,同时还具有神经递质、神经调质和神经营养等作用,使对PACAP的研究成为十分活跃的领域。PACAP属于血管活性肠肽（VIP）-胰高血糖素-生长激素释放因子-分泌素家族（Campbell and Scanes,1992）成员,已鉴别出包含27和38个氨基酸两种类型。对原索动物（McRory et al．,1997）、两栖类（蛙）（Alexandre et al．,2000）、爬行类（蜥蜴）（Pohland Wank,1998）、鸟类（鸡）（McRory et al．,1997）,啮齿类（鼠）（Ghatei et al．,1993）等脊椎动物PACAP的研究多集中在结构与进化方面,对功能了解甚少。     

斜带石斑鱼淋巴器官个体发育的组织学

本文应用连续组织切片技术和组织学观察,对出膜后1～60天的斜带石斑鱼(Epinephelus coioides)各期仔鱼、稚鱼和幼鱼的淋巴器官组织进行了研究,描述了淋巴器官的个体发育过程和组织学结构特征。研究表明：实验水温为22．0～27．8℃时,孵化后第10天出现头肾原基。头肾原基由未分化的造血干细胞组成。随着鱼体的生长,头肾原基的造血干细胞很快分化成不同类型的细胞;头肾主要由网状内皮系统支持下的淋巴造血组织构成。第11天出现脾脏原基。脾脏原基由造血细胞组成,淋巴化速度相对较慢。脾脏在整个发育过程中,红细胞和类红细胞占优势,没有红髓和白髓之分。第13天出现胸腺原基。胸腺发育速度较快,是明显的淋巴器官。胸腺主要由胸腺细胞(淋巴细胞)和上皮细胞组成,外区和内区没有明显的界限,但很容易区分。胸腺外被单层的上皮细胞层与咽腔相隔,保持浅表的位置,并且在整个发育过程中,胸腺与头肾是独立分开的。免疫器官原基出现顺序是头肾、脾脏和胸腺;而免疫器官淋巴化的顺序是胸腺,头肾和脾脏。和其它硬骨鱼类一样,斜带石斑鱼在早期发育阶段,淋巴器官的发育较迟,出现相对滞后的现象[动物学报49(6)：819～828,2003]。     

斜带石斑鱼CD4基因克隆和组织表达分析

CD4 作为TCR的共受体可以提高TCR/抗原-MHC复合体的稳定性,辅助TCR识别抗原,并且参与T细胞活化.本研究从斜带石斑鱼Epinephelus coioides头肾中克隆得到全长2240 bp的CDM cDNA序列,该序列包含长1410 bp的ORF,编码469个氨基酸,预测蛋白分子包含一段信号肽,4个Ig样区...     

斜带石斑鱼生长激素/催乳素家族基因cDNAs的分子克隆及其进化意义

从斜带石斑鱼垂体提取总。RNA,再取其50ng合成SMART cDNA。从所构建的垂体SMART cDNA质粒文库中筛选到生长激素／催乳素基因家族的2个成员的全长cDNA片段：生长激素(GH)基因全长为938bp,编码204个氨基酸;催乳素基因(PRI．)全长为1429bp,编码212个氨基酸。采用计算机软件Mega 2和CLUSTAL W1．64b对9种鱼的生长激素／催乳素基因家族的3个成员(GH、PRL和生长催乳素SL)的氨基酸序列进行系统分析,构建NJ分支系统树,对于序列中的插入／缺失位点则采用Pairaise Deletion,1000次自展(Bootstrap)分析计算各节点支持率。根据3个基因的氨基酸序列构建的系统树表明,石斑鱼与金头鲷、金鲈和牙鲆聚成一类,虹鳟与大马哈鱼聚成一类,鲫鱼与鲶鱼聚成一类,鳗鲡成另外一类。根据石斑鱼全长cDNA推断的氨基酸序列比较表明,SL相对GH和PRL有较高的保守性。石斑鱼的GH、PRL和SL的氨基酸同源性在24％～31％,但其C-端的氨基酸同源性较高,尤其是C-端的3个Cys是严格保守的。其中SL与GH的同源性(30．8％)高于与PRL的同源性(25．6％),GH和PRL的同源性最低(24．1％)。     

体重对斜带石斑鱼能量收支的影响

鱼类能量学是研究能量在鱼体内转换的学科,其核心问题之一是能量收支各组分之间的定量关系及其各种因子(如温度[1—5]、盐度[6]、体重[7—11]、性别[12]、摄食水平[13—18]、饵料种类[19,20]等)的影响作用。欧美等发达国家对鱼类能量学研究起步较早,迄今已经初步建立了多种鱼类的能量收支模式[21,22];国内在该领域较系统的研究起始于90年代初[23—25],主要局限于淡水鱼类,近年来又对海水鱼类进行了大量研究[26]。斜带石斑鱼(Epinephelus coioides)俗称青斑,为暖水性中下层鱼类,是广东省海水网箱养殖的主要品种之一。斜带石斑鱼分布于西太平洋的硫球群岛、澳大利亚以及贝劳和菲济群岛的东部,常栖息于大陆沿岸和大岛屿,但在河口和离岸100m深的水域中也可发现[27]。通过本项研究将有助于揭示海洋暖水性中下层鱼类的能量学特征。1材料与方法1·1材料来源与驯养实验用斜带石斑鱼,采自广东省大亚湾水产试验中心。实验用斜带石斑鱼经淡水浸泡10min后,置于室内容积0·5t桶内驯养,待摄食和生长趋于正常后,开始实验。驯化时间为30d。实验于2006年10—11月在广东省大亚湾水产试验中心进行。1...     

斜带石斑鱼(♀)×鞍带石斑鱼(♂)受精细胞学的组织切片观察

人工诱导雌核发育二倍化的关键是掌握极体排出和卵裂的时机,掌握该规律最直接的方式即进行受精细胞学的研究。以鞍带石斑鱼(Epinephelus lanceolatus)为父本,斜带石斑鱼(Epinephelus coioides)为母本,通过人工授精获得受精卵,收集受精后不同时刻的卵子,经Smith′s液固定、石蜡包埋、切片、苏木精-伊红染色和显微组织观察。结果表明:受精后第30~60 s精子入卵;第3 min精子星光出现;第5 min卵子排出第二极体;第7~15 min雌雄原核相互靠近,并最终融合;第15 min以后合子核开始分裂。鞍带石斑鱼精子进入斜带石斑鱼卵子的时间与多数硬骨鱼类比较没有显著差别;但是,第二极体排出及第一次卵裂的时间较早。研究展示了斜带石斑鱼与鞍带石斑鱼杂交的受精细胞学的基本发育过程,并为后续的人工诱导石斑鱼雌核发育研究提供了理论依据。     

斜带石斑鱼胸腺的显微和超微结构

本文通过解剖及组织切片技术、光学显微镜、透射和扫描电子显微镜技术,对斜带石斑鱼(Epinephelus coioides)胸腺器官组织进行了观察研究。结果表明:斜带石斑鱼胸腺实质主要由胸腺细胞(淋巴细胞)和网状上皮细胞构成。鱼体从Ⅰ龄之后,其胸腺发生明显的变化,与幼鱼有所不同,主要是胸腺可明显区分为三个区域:胸腺外皮质区、内皮质区和髓质区。外皮质区主要由网状上皮细胞、黏液细胞、成纤维细胞和少量淋巴细胞构成,细胞排列疏松;内皮质区主要由密集的淋巴细胞和网状上皮细胞组成,以含有大量的淋巴细胞为特征;髓质区主要由淋巴细胞和较多的网状上皮细胞构成,总体特征是淋巴细胞数量比内皮质区的少,且细胞排列较疏松。外皮质区、内皮质区相当于高等脊椎动物的皮质;髓质区相当于高等脊椎动物的髓质。髓质区之下有结缔组织,在Ⅱ龄以上的成体出现胸腺小体(Hassall's corpuscles)或类似胸腺小体的结构,而且随着年龄的增加,胸腺外皮质区增厚,结缔组织增加,还表现在内皮质区和髓质区组织逐渐萎缩变薄,胸腺的细胞组成类型和淋巴细胞数量上有所变化等等。这些现象在Ⅱ龄鱼开始出现,即胸腺呈现退化迹象,在Ⅲ龄以上鱼体呈现明显的退化和萎缩。胸腺表面扫描电镜结果表明:其上皮细胞表面具有微嵴以及由微嵴组成的指纹状结构,有一些微孔分布。透射和断面扫描电镜的结果进一步表明:胸腺组织内的细胞成分复杂,除了淋巴细胞和网状上皮细胞外,还具有巨噬细胞、肥大细胞、肌样细胞、浆细胞、指状镶嵌细胞和纤维细胞等。     

斜带石斑鱼(♀)×鞍带石斑鱼(♂)杂交子代(青龙斑)消化系统的早期发育

本文研究斜带石斑鱼(Epinephelus coioides)(♀)×鞍带石斑鱼(E.lanceolatus)(♂)杂交子代(青龙斑)仔、稚、幼鱼的消化系统发育,描述了其消化器官发育过程和组织学结构特征,充实青龙斑生物学研究文库,为其发育生物学研究和苗种培育提供技术支撑。青龙斑苗种培育于2012年6~8月期间进行。水温为(30±1)℃,盐度为28±1。利用形态学和连续组织切片技术,对出膜后0~40日龄幼鱼的消化系统进行了观察和研究。消化系统发育可划分为内源性营养、混合营养和外源性营养3个阶段:0至3日龄为内源性营养阶段,初孵仔鱼消化管为一简单的直形管,卵黄囊大,椭圆形,口和肛门尚未与外界相通;口腔中出现鳃弓的雏形,3日龄仔鱼食道由2~3层的复层立方上皮细胞组成,形成较低的褶皱;胃与小肠和食道的分界明显,上皮由单层柱状细胞组成;肠道分化,肛门开通体外,开始摄食;肝细胞团和胰腺细胞团形成。4~5日龄为混合营养阶段,6日龄之后进入外源性营养阶段,卵黄囊已经完全被吸收,前、中、后肠和直肠区分明显,肠黏膜上皮中出现少量的杯状细胞,由肠腔面向深层依次可以分为黏膜层、黏膜下层和浆膜层,肌层不明显。至25日龄,消化系统的结构和功能已经较为完善。38日龄时,胃、幽门盲囊、肠以及直肠各段分界明显,黏膜褶皱高度为前肠中肠后肠;肌层厚度为后肠前肠中肠;消化道和消化腺组织结构与成鱼基本相同。青龙斑的消化系统发育和分化是与其生理功能的逐步完善同步的。     

斜带石斑鱼L-氨基酸氧化酶基因克隆及表达分析

鱼类的L-氨基酸氧化酶(L-amino acid oxidase, LAAO)具有广泛的抑菌杀虫效果,为了解斜带石斑鱼(Epinephelus coioides)LAAO基因序列特征及其在刺激隐核虫(Cryptocaryon irritans)感染后的表达变化,该试验克隆得到2个石斑鱼LAAO基因:EcLAAO-1和EcLAAO-2,它们的ORF长度分别为1536和1569 bp,编码511和522个氨基酸,均含有氨基酸氧化酶(Amino_oxidase)结构域以及LAAO保守序列:DBM和GG motif。多重序列比对显示石斑鱼LAAO与其他鱼类LAAO具有较高的相似性。系统进化树分析表明,斜带石斑鱼的LAAO与硬骨鱼类亲缘关系较近。实时荧光定量PCR检测结果显示EcLAAO-1和EcLAAO-2在石斑鱼各组织均有表达,其中皮肤、鳃、胸腺、肝脏和肌肉中含量较丰富;在感染刺激隐核虫后,鳃和脾脏EcLAAO-1, EcLAAO-2表达量显著升高(P<0.05),这些结果暗示了石斑鱼LAAO参与先天性免疫,并在抗御刺激隐核虫感染中发挥重要作用。     

Androgens stimulate sex change in protogynous grouper,Epinephelus coioides: spawning performance in sex-changed males

We examined the efficacy of androgens (1.0 mg/kg body mass), testosterone (T), 11-ketotestosterone (11-KT), 17alpha-methyltestosterone (MT), testosterone propionate (TP) or androgen mixture (T, MT and TP in an equal ratio), for induction of sex change in protogynous orange-spotted grouper, Epinephelus coioides. The spawning performance in sex-changed males was also investigated. MT and androgen mixture at a dose of 1.0 mg/kg BW induced a sex transition and completion of spermatogenesis up to the functional male phase. The androgen mixture was most effective. Significantly, higher plasma T levels were found in MT and androgen mixture groups compared to control and other androgen implantation (T, TP or 11-KT) groups. We found that plasma levels of estradiol-17beta (E2) or 11-KT were not different among treated groups. Sex-changed males could successfully fertilize mature eggs. Fertilization and hatching rates were of 23.5-70.4% and 8.4-44.6%, respectively. The data demonstrated that induction of sex change by exogenous androgens in groups could apply to the aquaculture field for seed production.     

应用雄性激素诱导异育银鲫性转化的研究

用丙酸睾丸素溶液浸泡行雌核发育的异育银鲫(Allogynogenetic crucian carp)胚胎10～14天,诱导出了11.0％～13.6％的雄性鱼和10.5％～27.3％的兼性鱼。对50日龄的异育银鲫幼鱼投喂甲基睾丸素4个月,再继续饲养182～335天,诱导出了20％以上的雄性鱼。此外,部分实验鱼两侧卵巢的大小出现明显差异。Abstract：The embryos of allogynogenetic crucian carp were soaked in testosterone propionate for 10～14 days,11.0％～13.6％ fry were induced to be physiological males and 10.5％～27.3％ fry developed into bisexual sterile.The 50-day old fish were fed diet containing methyltestosterone for 4 months and then reared for 182～335 days.More than 20％ fry developed into males.In addition,the ovaries in both sides of some treated fish were significantly different in size.     

性逆转石斑鱼脑垂体差异表达基因克隆的筛选

以17α甲基睾丸酮投喂赤点石斑鱼(Epinephelusakaara),成功地获得了性逆转的有功能的雄鱼。用SMARTcDNA合成和长片段PCR技术构建了性逆转前后脑垂体cDNA文库;用抑制消减杂交技术建立了性反转前后抑制性差减文库。对获得的560个雄鱼脑垂体PCR阳性克隆和350个雌鱼脑垂体PCR阳性克隆进行斑点杂交,共筛选到103个差异表达cDNA片段。对其中29个克隆进行了测序,与GenBank中的已知基因序列进行同源性比较,发现有6个片段为促性腺激素α亚基前体基因(GenBank注册号:AY207430),与同属不同种的石斑鱼促性腺激素α亚基前体基因有97%的同源性;1个片段为生长激素前体(GenBank注册号:AY207431),与同属不同种的石斑鱼生长激素前体基因有99%的同源性;其余22个cDNA片段与GenBank中的序列无明显同源性 。     

Apoptotic cell death of oestrogen activated lactotrophs induced by tamoxifen

Stimulation and inhibition of lactotroph cells cause remarkable morphological and functional changes. In keeping with these changes, the size of the lactotroph cell population undergoes striking alterations due to proliferation or cell death. Factors involved in the induction of apoptosis of pituitary cells are not well established. We demonstrated earlier that oestrogens prevent lactotroph cells of female rats to die by apoptosis induced by bromocryptine treatment, a fact that can be reversed in ovariectomised rats. In this study, we developed experimental models for in vivo and in vitro studies to gain further insight on the survival effect of oestrogens on lactotrophs. In rats pretreated with oestrogens, tamoxifen generates a massive cell death by apoptosis as validated by the TUNEL technique and DNA electrophoresis of pituitary gland. On electron microscope observations, numerous lactotrophs exhibited progressive morphological changes in the nuclei compatible with the apoptotic process. The cells remaining intact also exhibit signs of inhibition due to a significant transformation of regular lactotrophs in atypical subtypes. In pituitary cell cultures exposed to tamoxifen and oestrogen simultaneously, most of the lactotrophs displayed features of apoptosis in the nucleus. The present reports gathered new evidences on the apoptogenic potential of tamoxifen on lactotroph cells, and corroborates the contribution of oestrogens to sustain both a balanced population of lactotrophs and a competent secretory activity. The concept that opposed activities, such as inhibition and stimulation, can activate apoptosis is also strengthen by these observations.     

4龄中华鲟垂体的EST分析

表达序列标签 (Expressed sequence tag, EST) 是鉴定基因表达规律和发现新基因的一种有效的分子生物学手段。为了能在中华鲟(Acipenser sinensis Gray) 中发现与生长和生殖内分泌调控相关的基因,我们构建了中华鲟垂体的SMART cDNA质粒文库。垂体是调节生长和生殖内分泌的重要器官。在本研究中,通过测序筛选得到了944个EST克隆,将所得EST 与 GenBank 数据库中的序列进行比对, 结果表明,802 (84.96%) 个克隆可以找到同源序列,共代表461个基因, 其中含132个已知功能基因;而 142 (15.04%) 个克隆不能找到同源序列。研究发现,在所有基因中,阿黑皮素原基因 (Proopiomelanocortin, POMC) 是出现次数最高的基因,占总EST数的10.17%, 显示出其在垂体中的重要地位。我们还发现了7个未知功能的基因并重点研究了其在心脏、肝脏、脾脏、肾脏、肌肉、精巢、卵巢和垂体等组织中的表达特异性。结果发现,4个基因：EG009334、EG009337、EG009338 和 EG009340为垂体特异性表达或垂体和卵巢特异性表达。对这些基因进一步的功能研究将有利于我们更好地了解中华鲟生长和生殖内分泌调控的分子机制。     

Die Feinstruktur gefriergebrochener Hormongranula

Zusammenfassung Gefriergebrochene Präparate (mit und ohne Ätzung) der Adenohypophyse und der C-Zellen der Schilddrüse von Ratte und Meerschweinchen wurden elektronenmikroskopisch untersucht. Bei den Hormongranula dieser Zellen verlaufen die Bruchflächen im allgemeinen zwischen den beiden Membranflächen oder zwischen Granulum-Inhalt und Membran. Nur relativ selten werden Granula quergebrochen. Auf beiden Hälften der gespaltenen Membranen der C-Zellgranula werden etwa mit gleicher Häufigkeit Proteinpartikel (100–200 Å) gefunden. Bei den Granula der somatotropen Zellen treten auf der dem Plasma anliegenden Membranhälfte deutlich mehr Proteinpartikel auf als auf der dem Granuluminhalt anliegenden Hälfte. Der Inhalt der somatotropen und C-Zellgranula erscheint bei dieser Präparationsmethode aus einer dichten Packung von 80–100 Å großen Partikeln zu bestehen.Eine besonders strukturierte Zone zwischen Membran und Granuluminhalt konnte bei den bisherigen Untersuchungen nicht festgestellt werden. Durch Ätzung der Gefrierbrüche ließen sich keine zusätzlichen strukturellen Details der Granula darstellen.Eine durch Auswertung von stereoskopischen Aufnahmen gewonnene Größenverteilungskurve für die C-Zellgranula wird vorgelegt.

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The fine structure of freeze-fractured hormone granules

Summary Freeze-fractured preparations (with and without etching procedures) of guineapig and rat thyroid (C-) cells and anterior pituitary (somatotropic-) cells have been investigated with the electron microscope. The hormone granules of these cells in general split either between the two lamellae of their unit membrane or between the granule contents and the unit membrane. Only rarely cross-broken granules have been observed. Inner and outer lamella of the unit membrane of the C-cell granules contain in more or less similar frequency moderate amounts of protein particles of 100–200 Å diameter. In case of the somatotrophs the outer lamella contains higher numbers of these particles than the inner one. The contents of the C-cell and somatotroph granules seems to consist of densely packed 80–100 Å particles. A particular zone between contents and membrane (as observed on micrographs with conventional electron microscopy) could not be detected on freeze-fractured preparations. The etching procedure does not reveal additional details of the granule structure.A size distribution curve of the C-cell granules as determined from stereo-pairs, is given.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Sex Differences in the Content of β-Endorphin and Enkephalin-Like Peptides in the Pituitary of Obese (ob/ob) Mice

Abstract: The β-endorphin content in pituitary extracts of male and female obese (ob/ob) and lean (+/?) mice was determined by radioimmunoassay. The amount of β-endorphin-like material contained in the pituitary of 3-month-old ob/ob male mice is similar to that of lean male mice. In contrast, the pituitary glands of female ob/ob mice have a greater amount of β-endorphin-like material than lean female mice. To determine with greater precision the molecular nature of the polypeptide that accounts for the increase in β-endorphin immunoreactivity, the various molecular forms of β-endorphin immunoreactivity were resolved by Biogel P-30 column chromatography. At least four peaks of immunoreactive material were detected. The first peak elutes in the void volume, and the second and the third peaks appear in the elution volumes of β-lipotropin and β-endorphin, respectively. That the material present in the void volume might be proopiocortin is supported by adrenocorticotropic hormone radioimmunoassay. The increased total β-endorphin immunoreactivity in pituitary glands of ob/ob mice is accounted for mainly by β-endorphin. The β-endorphin content of various brain structures of ob/ob mice is similar to that of lean littermates.     

Androgens and autistic traits: A study of individuals with congenital adrenal hyperplasia

Testosterone promotes male-typical neural and behavioral development in non-human mammals. There is growing evidence that testosterone exerts similar influences on human development, although the range of behaviors affected is not completely known. This study examined the hypothesis that autistic traits are increased following prenatal exposure to abnormally high levels of testosterone caused by congenital adrenal hyperplasia (CAH). Sixty individuals with CAH (34 female, 26 male) and 49 unaffected relatives (24 female, 25 male) completed the Autism Spectrum Quotient (AQ). Females with CAH scored significantly higher than unaffected females on total AQ score, largely due to enhanced scores on subscales measuring social skills and imagination. These results suggest that prenatal exposure to high levels of testosterone influences some autistic traits and that hormonal factors may be involved in vulnerability to autism.     

Annotation and analysis of expressed sequence tags (ESTs) from Chinese sturgeon (Acipenser sinensis) pituitary cDNA library

Chinese sturgeon Acipenser sinensis belongs to the family Acipenseridae, an ancient species of actinopterygian fishes. In order to advance molecular research on its reproduction, ontogenetic development, we were seeking for genomic information in the NCBI expressed sequence tag (EST database). We found 3384 indentified cDNA sequences which were assembled into 861 unigenes. Blast analysis revealed 301 unigenes shared high similarity with genes in the public databases, and these were classified into three groups: 202 known genes, 81 putative genes and 8 unknown genes. The remainder (560 genes) had no significant match to any protein sequence. Further, 255 unigenes and 333 unmatched unigenes were annotated with Gene Ontology (GO), which could be classified into cellular component, molecular function, and biological process. Among the known genes, the hormone genes pomc A (proopiomelanocortin), pomc B, GtH alpha I subunit (gonadotropin hormone), GtH alpha II subunit and GH (growth hormone) were present in this library. Comparison of the Chinese sturgeon proteins (GH, GtH alpha subunit and POMC) to proteins of other species showed higher levels of homology among sturgeon species. We performed five hormone related genes including GnRHRI (gonadotropin-releasing hormone receptor I), cpH (carboxypeptidase H), ppiB (peptidylprolyl isomerase B), stmn3 (stathmin-like 3), 7B2 (neuroendocrine protein 7B2), and four novel genes (contig 192, 177, 170 and 168) a semi-quantitative RT-PCR on different tissues from Chinese sturgeon.     

Relationship between 22-kHz calls and testosterone in male rats

Ultrasonic calls in rats induced by the presence of a predator, referred to as “22-kHz calls,” are mainly emitted by socially dominant male rats. Testosterone levels are closely related to social dominance in male rats. In the present study, we investigated the relationship between the emission of stress-induced 22-kHz calls and circulating testosterone levels in male rats, using a combination of surgery (castration or sham operation) and chronic steroid administration (testosterone or cholesterol) to modify circulating testosterone levels. We also assessed the effects of androgen and/or estrogen receptor antagonists on the emission of 22-kHz calls in male rats. An air puff stimulus, known to reliably induce 22-kHz calls in rats, was used as a stressor. Castrated rats with cholesterol implants exhibited significantly fewer 22-kHz calls than rats that had received a sham operation and cholesterol implants, and there was no significant difference between castrated rats with testosterone implants and rats that had received a sham operation and cholesterol implants. Only male rats pretreated with a binary mixture of androgen and estrogen antagonists exhibited significantly fewer 22-kHz calls than controls. These results show that testosterone in male rats has a positive effect on the emission of stress-induced 22-kHz calls, and the calls may be regulated by the activation of both androgen and estrogen receptors.