微囊藻毒素在鲤体内富集的初步研究

为了解微囊藻毒素MC-LR在鲤各组织器官中的生物富集作用,采用腹腔注射法将MC-LR纯品稀释液(0.215μg/g)注射到鲤体内,酶联免疫吸附法(ELISA)检测0、1、3、12、24和48 h时肾脏、肝脏、肌肉、胆囊、空肠和卵巢中MC-LR含量分布和积累规律.结果显示,MC-LR在肾脏含量最高,均值为(1.007±0.120)μg/g(各器官均按干重计),其次是肝脏(0.490±0.060)μg/g、胆囊(0.355 ±0.011) μg/g、空肠(0.210±0.005)μg/g、卵巢(0.082±0.021)μg/g和肌肉(0.047±0.003) μg/g.鲤不同组织器官对MC-LR的富集能力存在较大差异,肾脏是MC-LR的主要靶器官,卵巢中也存在少量MC-LR富集,肌肉对MC-LR的生物富集量远低于其他组织器官.试验48 h时,鲤体内各组织器官中和鱼缸水中MC-LR含量均有一定程度的下降,表明鲤体内对MC-LR有较强的解毒机制.     

MrFoxl2-dsRNA对罗氏沼虾卵巢中foxl2及生殖相关基因表达的影响

foxl2是性腺分化、发育和维持等发育过程的转录调节因子,在一些哺乳动物和鱼类的卵巢发育中发挥了重要作用。本研究比较分析了性成熟前后罗氏沼虾(Macrobrachium rosenbergii)foxl2基因在肝胰腺与卵巢中的表达,并利用RNA干扰技术沉默MrFoxl2(M.rosenbergii foxl2)基因后分析其在卵巢成熟过程中的功能。荧光定量结果显示,MrFoxl2在卵巢组织中高表达,并且性成熟后MrFoxl2在卵巢中的表达较性成熟前显著提高。在注射MrFoxl2-dsRNA后MrFoxl2基因的表达显著下降,并且使用1μg/g的注射剂量对MrFoxl2基因的敲降效果最佳。此外,MrFoxl2基因沉默后,生殖相关基因VG与EcR的表达显著下调,CatL基因表达显著上调。研究结果表明,MrFoxl2在罗氏沼虾卵巢成熟过程中可能发挥了重要作用。     

CpG寡脱氧核苷酸对中华绒螯蟹非特异性免疫指标的影响

细菌DNA和人工合成的含CpG基序的未甲基化寡脱氧核苷酸(ODNs)均能刺激机体的免疫反应.本研究将鱼的免疫刺激剂寡脱氧核苷酸序列ODN-1670(含1个CpG基序)和ODN-R(含1个反向CpG基序,即Gpc)按1μg/kg、5μg/kg、25μg/kg的剂量对中华绒螯蟹(Eriocheir sinensis)进行体腔注射,同时以注射0.1mL生理盐水组与未进行任何处理的空白组作为对照.在注射前(0d)和注射后第1天、3天、5天、8天对血细胞呼吸爆发活性(O2-产量)、过氧化物酶(POD)活性、超氧化物歧化酶(SOD)活性进行测定.结果显示,注射25μg/kg ODN-1670能显著增强实验蟹血细胞的呼吸爆发活性(P＜0.05),注射1μg/kg、5μg/kg、25μg/kg剂量的ODN-1670能显著增强实验蟹血清的POD、SOD活性(P＜0.05).而注射3种剂量的ODN-R对血细胞呼吸爆发活性、POD活性和SOD活性均没有显著影响(P＞0.05).本实验结果表明,注射适量ODN-1670能增强中华绒螯蟹的非特异性免疫功能.     

玻璃化冻存对斑马鱼卵母细胞的影响

采用比色法测定了在不同玻璃化冻存液中冻存1、2、4、8d时斑马鱼卵母细胞中谷胱甘肽含量和三磷酸腺苷酶的活性,以研究玻璃化冻存对卵母细胞谷胱甘肽含量和线粒体三磷酸腺苷酶活性的影响。试验结果显示,V4组斑马鱼(玻璃化液组分是:1.5mol/L甲醇、6.0mol/L乙二醇、0.25mol/L蔗糖)卵母细胞冻存1、2、4、8d后存活率最高[分别为(48.89±2.58)%、(39.35±1.34)%、(24.18±2.32)%和(14.56±1.64)%],显著高于其他组。经玻璃化冻存后,斑马鱼卵母细胞谷胱甘肽含量和线粒体三磷酸腺苷酶活性均随冻存天数的增加而显著下降(P0.05)。V4组的谷胱甘肽含量[(13.37±0.59)mg/g]和线粒体三磷酸腺苷酶活性较高,线粒体Ca~(2+)-三磷酸腺苷酶[(3.823±0.144)U/mg]、Mg~(2+)-三磷酸腺苷酶[(4.503±0.144)U/mg]和Na~+/K~+-三磷酸腺苷酶[(7.520±0.198)U/mg]活性最高。各组细胞的活性随着冻存天数的增加而逐渐下降。试验结果表明,玻璃化冻存不能完全避免低温和冷冻保护剂的毒性和对卵母细胞的损伤。     

施氏鲟促卵泡激素受体基因cDNA的克隆、表达及调控

为研究促卵泡激素受体基因(FSHR)在施氏鲟(Acipenser schrenckii)性腺分化过程中的表达、分布及调控作用,采用反转录聚合酶链式反应(RT-PCR)和RACE技术克隆得到施氏鲟促性腺激素(FSHR)基因的全长cDNA序列,并对其编码氨基酸序列的特征、基因表达及调控作用进行了分析。结果显示:施氏鲟FSHR基因的cDNA全长为2696 bp,编码661个氨基酸,属于含有信号肽、跨膜结构且分泌到细胞外的疏水性蛋白。氨基酸同源性及进化分析表明,施氏鲟与小体鲟(A.ruthenus)FSHR序列一致性最高,亲缘关系最近。PCR结果显示,FSHR mRNA的表达具有广泛性,各组织中均有表达,但在性腺、垂体中的表达量较高,显著高于其他组织中的表达量。LHRH-A注射实验显示:不同浓度组中,施氏鲟性腺组织中FSHR mRNA的表达量呈先升高后降低的变化趋势;与对照组相比,1μg/kg组在诱导3 d时性腺组织中FSHR mRNA的表达量达到最高,极显著高于3μg/kg和5μg/kg组。与性腺组织中FSHR mRNA表达模式不同,垂体中FSHR mRNA的表达呈升高后降低趋势,3μg/kg组在注射后第3天达最大值。综上结果表明,LHRH-A可通过调控FSHR的表达参与施氏鲟的早期性腺发育。     

红鳍东方鲀伪雌鱼卵巢发育迟滞的调控机制

伪雌鱼的培育是红鳍东方鲀(Takifugurubripes)全雄制种技术研发的关键环节之一,然而外源雌激素诱导获得的伪雌鱼表现出卵巢发育迟滞,降低了其育种价值和效率。为探讨红鳍东方鲀伪雌鱼卵巢发育迟滞的调控机制,本研究从孵化后20日龄开始,用10μg/L 17β-雌二醇(E2)浸泡红鳍东方鲀稚幼鱼,每天浸泡1次,每次2 h,至90日龄结束。在90、180和330日龄分别采集处理组(10μg/L E2)遗传雄性幼鱼和对照组(0μg/L E2)遗传雌性幼鱼,比较两组幼鱼性腺的组织学和形态学变化特征、下丘脑-垂体-性腺轴相关激素(FSH、LH、E2和17α, 20βOH-PROG)和基因(fshr、lhr、erα、erβ1、erβ2)及脂质积累相关基因(lpl和vldlr)的变化规律。结果显示:10μg/LE2可将遗传雄性幼鱼全部诱导为伪雌鱼,且伪雌鱼直至330日龄未二次反转为间性或者雄性,但其性腺系数、卵母细胞数量及卵黄生成前期的卵母细胞面积均显著小于对照雌鱼。此外, 90日龄伪雌鱼的lhr和pgr的表达量显著高于同期对照雌鱼,而17α,20β-PROG的含量及fshr的表达量显著低于对照组;180日龄伪雌鱼的vldl表达量显著低于对照组;330日龄伪雌鱼的激素含量及基因表达量没有显著差异。综合分析伪雌鱼性腺发育的形态学、组织学和性腺轴相关激素及基因变化规律可见,足够浓度的外源E2能够诱导并维持伪雌鱼的卵巢特征,但E2浓度过高,一方面可能抑制fshr和vldlr基因的表达,从而影响脂质在卵黄生成早期卵母细胞中的积累,导致红鳍东方鲀伪雌鱼卵母细胞生长迟缓;另一方便,高浓度E2抑制伪雌鱼卵原细胞减数分裂的启动,是导致红鳍东方鲀伪雌鱼卵母细胞数量较少的原因之一。     

养殖中华鲟性腺发生与分化的组织学研究

用组织切片方法研究人工繁育中华鲟的性腺发生及两性分化过程。中华鲟出膜后3d，原始生殖细胞以单细胞的形式存在于肾管区腹下方。出膜后11d，生殖褶形成，到2月龄和7月龄时，性腺中分别出现血管和脂肪组织，直到8月龄性腺均处于两性未分化状态，划分为0期。9月龄(体重0．6～1．1kg)，性腺出现组织学水平上的两性分化，性腺划入I期。I期性腺中，精(卵)原细胞进行有丝分裂。以初级精(卵)母细胞出现作为Ⅱ期开始的标志，精巢和卵巢分别于1．8～2．2年龄(1.55～5．6kg)和2．5～3．0年龄(3．1～8．2kg)进入Ⅱ期。3年龄以后，性腺发育分化陆续达到肉眼可分辨性别的程度，5～5．6年龄(18～35．5kg)时，所有的性腺能肉眼区分雌雄，但精巢和卵巢仍都处于Ⅱ期，其中卵母细胞的卵径为60～240μm。     

Nesfatin-1调控斜带石斑鱼生殖作用的研究

Nesfatin-1是核连蛋白-2(nucleobindin-2,NUCB2)裂解后产生的一种神经肽。本研究克隆得到斜带石斑鱼两种编码NUCB2(NUCB2a和NUCB2b)基因的c DNA序列(nesfatin-1a和nesfatin-1b),并对它们的表达模式进行了研究。结果表明nesfatin-1a和nesfatin-1b在脑组织中广泛表达。在下丘脑中,两种nesfatin-1在卵巢发育的Ⅰ-Ⅴ时期表达量都比较低,在Ⅵ时期卵巢(退化的卵黄生成阶段的卵母细胞)中的表达量显著升高,揭示nesfatin-1可能参与了卵巢成熟的调控。进一步采用腹腔注射实验研究了两种nesfatin-1对生殖轴的影响。注射nesfatin-1a和nesfatin-1b 15 min和60 min后,下丘脑中促性腺激素释放激素Ⅰ型(Gn RH-Ⅰ)和促性腺激素释放激素Ⅲ型(Gn RH-Ⅲ)的表达量都显著下降;此外,垂体中FSH-β的表达在注射nesfatin-1 60 min后显著下调。这些结果表明nesfatin-1a和nesfatin-1b参与了斜带石斑鱼下丘脑-垂体-性腺轴(HPG)的调控。     

海南岛糙海参性腺发育的组织学特征

对中国海南岛糙海参(Holothuria scabra)的性腺发育过程进行了组织学观察。结果表明,海南糙海参的性腺发育可分为恢复期(Ⅰ期)、增长期(Ⅱ期)、成熟期(Ⅲ期)、部分排放期(Ⅳ期)、排空期(Ⅴ期)共5个时期。1)精巢发育特征:Ⅰ期,生殖上皮出现精原细胞和精母细胞,生殖管壁最厚;Ⅱ期,生殖管上皮变薄,褶皱往管腔中延伸;Ⅲ期,成熟精子充满整个生殖管;Ⅳ期,生殖上皮恢复褶皱形态;Ⅴ期,生殖管中剩余少量残留精子。2)卵巢发育特征:Ⅰ期,在生殖上皮附近附着卵原细胞和卵黄发生前期卵母细胞;Ⅱ期,大量卵黄发生时期卵母细胞与卵原细胞及卵黄发生前期卵母细胞共存于生殖管中;Ⅲ期,卵黄发生时期卵母细胞充满生殖管;Ⅳ期,在生殖上皮附近重新出现卵黄发生前卵母细胞,生殖管腔中出现营养性吞噬细胞;Ⅴ期,生殖管腔残留少数卵细胞。     

紫贻贝产卵前后转录组分析与相关基因的筛选

为探究紫贻贝(Mytilus galloprovincialis)雌性生殖相关基因在产卵过程中发挥的功能及其参与产卵活动的调控机制,对紫贻贝产卵前后的性腺组织进行了转录组测序分析。结果显示,紫贻贝产卵前后存在的差异表达基因为4060个,其中上调的基因数目为1488个,下调的基因数目为2572个。GO富集分析显示,这些差异基因主要参与生长、发育过程、生殖、生殖过程等生殖相关的生物学途径。KEGG富集分析显示,与产卵、生殖相关的信号通路包括TCA循环、GnRH信号通路、卵巢类固醇生成、雌激素信号通路、催产素信号通路、类固醇生物合成等能够被显著富集。荧光定量分析结果显示,促黄体生成激素β (LHβ)和基质金属蛋白酶14 (MMP14)基因表达水平显著升高,促性腺激素释放激素受体(GnRHR)、细胞色素P4503A酶(CYP3A)、细胞色素P45017A酶(CYP17A)基因表达水平显著下降。本研究利用转录组学分析了紫贻贝在产卵前后性腺组织转录水平的差异,揭示了紫贻贝性腺组织中参与调节产卵过程的主要途径和候选基因,为进一步阐明紫贻贝产卵活动的内在调控机制提供数据支撑,也为紫贻贝苗种繁育提供理论...     

太湖微囊藻毒素在罗非鱼体内的累积及生物降解的初步研究

为了解蓝藻水华期间微囊藻毒素在罗非鱼体内的分布及累积传递过程，2008年6月至8月采集了高密度蓝藻池塘及太湖网箱内的鱼样及水样，用ELISA法对鱼样和水样进行微囊藻毒素MC-LR含量的检测。结果表明：池塘水体微囊藻毒素MC-LR含量变化范围在0.123～0.514ug/L间，MC-LR含量随着藻密度的下降而降低，对照组水体MC-LR浓度显著高于实验组MC-LR含量。池塘鱼体肌肉组织微囊藻毒素MC-LR累积含量在1.194～3.615ng/g间，肝脏组织微囊藻毒素MC-LR累积含量显著高于肌肉组织。将池塘与网箱罗非鱼转至无微囊藻水体中暂养，跟踪检测MC-LR含量变化，池塘和网箱鱼体肌肉组织微囊藻毒素MC-LR含量均低于人体每日可耐受摄入量，而肝脏组织藻毒素MC-LR含量则分别需要经过10～20天自然生物降解后降低至安全摄入量之下。并讨论了微囊藻毒素在鱼体内的组织分布与食物链中的累积传递。     

Effect of Luteinising Hormone-Releasing Hormone Analogue and Human Chorionic Gonadotropin on Ovulation, Plasma and Ovarian Levels of Gonadal Steroids in Greenback Flounder Rhombosolea tapirina

Female greenback flounder Rhombosolea tapirina were treated with either des Gly¹⁰ [D-Ala⁶] LHRH ethylamide (LHRH-a) at 50 or 100 pg/kg intraperitoneal injection (ipi), LHRH-a cholesterol pellet (LHRH-a pellet) 100 μg/kg implanted intraperitoneally, or human chorionic gonadotropin (hCG) 1,000 IU/kg ipi. Treatment with hCG, LHRH-a (50 μg/kg) ipi and LHRH-a pellet increased the total number of ovulations above control levels. LHRH-a (100 μg/kg) ipi was not consistently more effective than the control and both LHRH-a (50 μg/kg) ipi and LHRH-a pellet induced more ovulations than LHRH-a (100 μg/kg) ipi. Of those fish that ovulated, most ovulated more than twice, and most ovulations occurred at daily intervals. Oocyte diameters increased and oocyte stage advanced significantly in response to all exogenous hormone treatments. This was accompanied by increases in plasma and ovarian levels of 17β-estradiol (E₂, and in most cases, plasma and ovarian levels of testosterone (T). Plasma and ovarian levels of 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) were not consistently elevated in association with reproductive events.     

In situ studies on the bioaccumulation of microcystins in the phytoplanktivorous silver carp (Hypophthalmichthys molitrix) stocked in Lake Taihu with dense toxic Microcystis blooms

The phytoplanktivorous silver carp is an important biomanipulation fish to control cyanobacterial blooms and is also a food fish with the greatest production in China. The accumulation of the hepatotoxic microcystins (MCs) determined by LC-MS in various organs of silver carp was studied monthly in Lake Taihu dominated by toxic Microcystis aeruginosa. Average recoveries of spiked fish samples were 78% for MC-RR and 81% for MC-LR. The highest content of MCs was found in the intestine (97.48 μg g^− 1 DW), followed by liver (6.84 μg g^− 1 DW), kidney (4.88 μg g^− 1 DW) and blood (1.54 μg g^− 1 DW), and the annual mean MC content was in the order of intestine > liver > kidney > blood > muscle > spleen > gallbladder > gill. Silver carp could effectively ingest toxic Microcystis cells (up to 84.4% of total phytoplankton in gut contents), but showed fast growth (from 141 g to 1759 g in 1 year in mean weight). Silver carp accumulated less microcystins in liver than other animals in the same site or other fish from different water bodies at similar level of toxin ingestion. There was possible inhibition of the transportation of the most toxic MC-LR across the gutwall. Muscle of silver carp in Lake Taihu should not be consumed during period of dense Microcystis blooms while viscera were risky for consumption in more months.     

Ovarian histology of stunted pond-reared Macrobrachium rosenbergii females

The present study describes the ovarian histology of stunted freshwater prawn Macrobrachium rosenbergii. The ovarian maturation of stunted animals was examined and compared with similar‐aged normal females. Ten animals of the stunted group and each maturation stage of the normal group were sampled from the same pond and had their ovaries removed for histological analysis. Body weight, body length, ovarian weight and gonadosomatic index (GSI) were recorded for each female. The diameters of the different oocyte types were compared among groups through histological assessments. The ovarian histology of stunted M. rosenbergii females indicated that although the somatic growth is severely affected (7.6 g), some energy has been placed on the vitellogensis. Stunted females showed the simultaneous occurrence of previtellogenic, vitellogenic and mature oocytes in their ovarian tissue, but overall oocyte diameter and GSI (1.02%) were significantly affected when compared with normal females.     

Ultrastructural alteration of lymphocytes in spleen and pronephros of grass carp (Ctenopharyngodon idella) experimentally exposed to microcystin-LR

Relatively little is known in relation to pathological changes of immune organs in fish when exposed to MC-LR. The ultrastructural alteration of lymphocytes was examined in the spleen and pronephros of grass carp Ctenopharyngodon idella injected experimentally with microcystin-LR. The fish were intraperitoneally injected with MC-LR at a dose of 50 µg/kg body weight, and the spleen and pronephros were dissected out at 1, 2, 7, 14 and 21 days post intraperitoneal injection (dpi). Pathological changes were then examined by transmission electron microscopy. Apoptosis was detected only in lymphocytes in the spleen, with obvious apoptotic features observed at 2 dpi; pathological changes of lymphocytes in the pronephros were also serious with mitochondria being highly edematous. However, damaged lymphocytes were almost un-observed in the spleen and pronephros at 21 dpi. These findings suggest that MC-LR can induce toxic effect on immune organs in grass carp, and the spleen may be much more sensitive to MC-LR stimulation.     

Annual changes in ovarian development and plasma estradiol-17β level in reared female common Japanese conger, <Emphasis Type="Italic">Conger myriaster</Emphasis>

ABSTRACT:   Annual changes in the gonadosomatic index (GSI), oocyte diameter, gonadal histology and plasma estradiol-17β (E₂) levels were investigated in female common Japanese conger Conger myriaster in captivity. Juveniles were caught in September 1999 and reared in seawater at temperatures ranging from 10–20°C for 3 years. All fish were immature when captured in September 1999. GSI and oocyte diameter increased from October 2000, peaked between summer and autumn 2001, and bottomed-out in winter 2001. Plasma E₂ level also increased from October 2000, but remained high. The ovarian developmental stage was at the peri-nucleolus stage or the oil droplet stage until September 2000. Vitellogenesis started in October 2000 and oocytes progressed to the tertiary yolk globule stage by summer 2001. However, vitellogenic oocytes regressed in all females after autumn 2001. The neogenetic oocytes began to increase after November 2001 and ovarian development progressed in 2002 as they did in 2001, although maximum GSI in 2002 was half its 2001 value. These data indicate that ovarian development in the common Japanese conger has an annual periodicity, and that these congers may be able to spawn in multiple years under rearing condition.     

Erythrocyte damage of crucian carp (Carassius auratus) caused by microcystin-LR: in vitro study

Fish suffer from anemia and hypovolemic hypotensive shock after in vivo exposure with microcystins. However, except for in vivo causes for anemia and hypotension, an in vitro study of fish erythrocytes exposed to MC is necessary. For a better understanding of hematology toxicity of MC, the main aim of the present study was to investigate the toxic effects of microcystin on fish erythrocytes in vitro. Crucian carp erythrocytes were incubated in vitro with microcystin-LR (MC-LR) at doses of 0, 1, 10, 100 and 1,000 nM. The level of lipid peroxidate significantly increased in MC-LR treatment groups. Glutathione decreased after exposure to MC-LR. The activities of antioxidative enzymes, including superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase, were significantly increased after exposure with MC-LR. The hemolysis was significantly increased, while the activities of acetylcholinesterase, Na⁺–K⁺-ATPase and Ca²⁺–Mg²⁺-ATPase were significantly decreased. In addition, pathological alterations in agglomerated and jagged erythrocytes were observed in blood smears. The findings indicate that damages to erythrocytes should also be responsible for anemia and hypotensive shock or even death.     

Molecular endocrinology of oocyte growth and maturation in fish

Pituitary gonadotropins (GTHs) are of primary importance in triggering oocyte growth and maturation. However, the actions of GTHs are not direct, but are mediated by the ovarian production of steroidal mediators of oocyte growth (estradiol-17β) and maturation (maturation-inducing hormone, MIH; 17α,20β-dihydroxy-4-pregnen-3-one, 17α,20β-DP in salmonid fishes; 17α,20β,21-trihydroxy-4-pregnen-3-one, 20β-S in sciaenid fishes). It is established that production of estradiol-17β and 17α,20β-DP by salmonid ovarian follicles occurs via the interaction of two cell layers, the thecal and granulosa cell layers (two-cell type model). A distinct shift in the salmonid steroidogenesis from estradiol-17β to 17α,20β-DP occurs in the ovarian follicle layer immediately prior to oocyte maturation. It is possible that this shift is a consequence of dramatic changes in the expression of the genes encoding various steroidogenic enzymes. As an initial step to address this question, we have isolated and characterized the cDNAs encoding a number of ovarian steroidogenic enzymes including the rainbow trout cholesterol side-chain cleavage cytochrome P-450, 3β-hydroxysteroid dehydrogenase (HSD), 17α-hydroxylase/17,20 lyase cytochrome P-450, aromatase cytochrome P-450 cDNAS as well as the pig 20β-HSD cDNA. Estradiol-17β stimulates the hepatic synthesis and secretion of a yolk precursor, vitellogenin. Vitellogenin is then transported to the ovary where it is selectively taken up into the oocyte by a receptor-mediated process involving specific cell-surface receptors. Estradiol-17β was also shown to induce the synthesis of egg membrane proteins in the liver. The maturation-inducing action of 17α,20β-DP and 20β-S is through the binding to the oocyte plasma membrane. This initial MIH-surface interaction is followed by the formation of the major mediator of MIH, maturation-promoting factor (MPF). We have purified MPF from mature oocytes of carp. Carp MPF consists of two components: the homolog of the cdc2⁺ gene product of fission yeast (p34^cdc2) and cyclin B. The cdc2 kinase protein is present in immature oocytes as well as in oocytes induced to mature by 17α,20β-DP treatment, while cyclin B proteins can be detected only in mature oocytes. Addition of bacterially expressed goldfish cyclin B to the extracts of immature goldfish oocytes induced MPF activation. These results suggest that the appearance of cyclin B protein is a crucial step for 17α,20β-DP-induced oocyte maturation in fish.     

Stimulation of spermiation and induction of ovulation in pike (Esox lucius)

Mature northern pike were given various hormonal treatments in March or April in order to stimulate spermiation or to induce ovulation. In males the total amount of sperm collected after treatment increased, in comparison with saline-injected males, by 3–11 times with partially purified salmon gonadotropin (PPSG-activity: half of the highly purified s-GTH; injected at doses of between 5 and 100 μg/kg body weight); 3–6 times with crude carp pituitary extract (0.5–3 mg/kg body weight); and 3–7 times with fresh pike pituitaries (14 and 1.2 mg wet weight/kg body weight). The sperm obtained after hormonal treatment was of good quality. Intracardiac injection of superactive LRH analogue had no effect. In females, PPSG induced 90 and 100% ovulation at the doses of 50 and 25 μg/kg body weight. Dried salmon pituitaries (2.5 mg/kg, equivalent to 50 μg of PPSG) gave 25% ovulation; at 10 mg/kg, 25% complete ovulation was again recorded, but in addition 70% of the females showed oocyte maturation and partial ovulation. Similarly, dried carp pituitary (3 mg/kg) induced only oocyte maturation but no ovulation. The oocytes obtained after hormonal treatment were in general fertile. Intraperitoneal injection of LRH in an emulsified form induced neither oocyte maturation nor ovulation. The lack of effect of LRH analogue is discussed and shows that the use of this compound as a substitute for pituitary preparation is not very promising.