七星瓢虫成虫脂肪体核酸代谢与生殖滞育的关系

本文以微量萤光法测定七星瓢虫成虫繁殖期、滞育期脂肪体核酸、蛋白含量的变化,结果表明:繁殖期雌虫卵巢发育Ⅰ和Ⅱ级时RNA/DNA比值分别为2.47和4.50;卵巢发育至Ⅲ级时RNA/DNA比值出现高峰为8.44,Ⅳ级时降为3.60,产卵后显著下降至1.76。所以脂肪体RNA/DNA比值可做为卵黄蛋白合成的一项指标。 成虫羽化后,以短光照(每日9小时光照,15小时黑暗)诱导成虫滞育,测定预滞育期RNA/DNA比值及蛋白质含量分別为3.16及59微克/毫克脂肪体;进入滞育期则分别降为1.24和14微克/毫克脂肪体;滞育结束后RNA/DNA比值上升为7.04,蛋白含量为133微克/毫克脂肪体。 成虫繁殖期与滞育期不同阶段脂肪体核酸、蛋白量的差异反映卵黄蛋白合成水平。核酸蛋白质含量低可做为滞育出现的信号。而滞育结束后RNA/DNA及蛋白含量的增加,可做为滞育结束的一个生化指标。     

光周期对三带喙库蚊发育期间体内核酸动态的影响

作者测试了长短光周期处理的三带喙库蚊Culex tritaeniornynchus(Giles)各虫态与滞育及解滞蚊体内RNA与DNA的变化.结果表明:幼虫与蛹核酸含量最高,成虫期变化比较稳定.短光周期可以降低该蚊发育期间核酸的合成作用.雌蚊卵形成期间DNA相对稳定,吸血后卵巢开始发育,RNA含量明显增加,36—48小时达到高峰,然后逐渐下降.10—15日龄的滞育蚊体内DNA比发育蚊低,解滞蚊RNA与DNA含量均比发育蚊为高.新蚊、发育、滞育与解滞蚊间DNA、RNA含量不同,DNA、RNA减少与增加可用作判断滞育发生与解除的一个生化指标.     

家蚕后丝腺亚细胞组分中核酸和蛋白质的分布

丝心蛋白是家蚕后丝腺的分泌产物。在家蚕五龄早期,后丝腺细胞中迅速形成丝心蛋白合成所必需的一些亚细胞结构,在后丝腺细胞质中粗糙内质网迅速增多,平滑内质网迅速形成空泡状或管状。而且生化测定表明,在五岭早期后丝腺中DNA,RNA、蛋白质和脂肪的含量皆迅速增加,这些为在五龄后期丝心院的合成,即为丝心蛋白信息的转录和翻译,提供亚细胞结构和生化代谢的物质基础(Tashiro等1968;赤井弘1973)。因此有必要对家蚕后丝腺亚细胞组分如细胞核、线粒休、微粒体和105,000g上清液中核酸和蛋白质代谢进行研究。本文初步建立一种分离家蚕后丝腺亚细胞组分的差级离心方法,并测定各亚细胞组分中核酸和蛋白质的含量。     

蓖麻蚕蛹卵巢发育过程中亚细胞组分核酸和蛋白质的代谢与脑激素的影响

在20℃条件下,蓖麻蚕蛹期发育需14天,接近发育完成的卵巢中各亚细胞组分中DNA、RNA和蛋白质含量迅速增长。经手术去脑后的蓖麻蚕无脑蛹卵巢不发育,其卵巢中上述成分的含量皆处于蛹早期水平。经脑激素处理后的无脑蛹卵巢即可迅速发育,在处理后18—20天,卵巢发育可接近正常水平,但较晚。在电镜下观察到的游离多核糖体呈二到七聚体,其紫外光谱特性为A260/A280=1.70;A240/A280=1.27。将脑激素注射到无脑蛹中去,可促使卵巢中游离多核糖体含量的增加。     

东亚飞蝗生殖的研究:雌蝗成虫卵巢发育过程中核酸和蛋白质的代谢与激素调节

本文报道东亚飞蝗交配后雌蝗卵巢中核酸和蛋白质的含量变化,以及孤雌生殖和雄蝗促性腺因子对卵巢中核酸和蛋白质代谢的影响。雌蝗羽化后7—10天进行交配,卵巢开始迅速发育,卵巢鲜重、总磷量以及蛋白质含量皆迅速增长,同时末端卵母细胞长度亦不断增加。到羽化第15天时卵巢已接近发育完成。末端卵母细胞长达6.2毫米。在各种磷化合物中,酸溶性部分在羽化15天时占总磷量的70%,磷脂磷可达20%。这表明酸溶性磷化合物和磷脂在卵巢发育过程中有较高的代谢和积累。卵巢中RNA-P增长28倍,DNA-P,增长6倍。RNA/DNA比值随着卵巢的发育而增加,这标志着蛋白质在卵巢中合成;对蛋白质含量的测定也证实了这一点。如果以RNA-P和DNA-P占总磷量的百分含量或以每百毫克卵巢鲜重计算其含量,则在卵巢发育过程中反而皆相对降低,表明卵巢中其他含磷化合物的积累优于核酸磷的增长。当雌蝗第一次产卵后,卵巢的各组成成份迅即减少,此后四天内卵巢中核酸和蛋白质的含量复可再度迅速增长,末端卵母细胞(即原未产卵前之末端第二卵母细胞)亦进一步长大,从而表现了卵巢发育的周期性变化。 人为隔离的孤雌生殖的雌蝗在羽化后40天内,卵巢发育缓慢,其末端卵母细胞长度增长缓慢,卵蜒中核酸和蛋白质的含量皆较低,相当于正常发育卵巢5—10天的水平。如将雄蝗脂肪体脂类提取物涂抹于羽化后5—7天雌蝗体表,则表现出促进雌蝗卵巢发育的作用,卵巢鲜重、卵巢中核酸等磷化合物和蛋白质含量,以及卵管中末端卵母细胞长度,在短时期内,基本上相同于正常交配对照的卵巢发育水平。因此证实:雄媓促健腺因子对卵巢发育过程中核酸和蛋白质代谢和合成起调节作用。这种调节因子可能是与保幼激素共同起作用的。     

柞蚕蛹滞育和滞育解除过程中海藻糖酶基因表达模式及酶活力分析

【目的】克隆柞蚕Antheraea pernyi海藻糖酶(trehalase,Treh)基因,探讨该基因在柞蚕蛹滞育和滞育解除过程中的表达模式与海藻糖酶活力变化,为阐明柞蚕蛹滞育期间糖代谢机制提供参考。【方法】利用RT-PCR技术从柞蚕蛹中克隆获得海藻糖酶基因,并对其进行生物信息学分析。采用半定量RT-PCR检测长光照(17L∶7D)处理后的滞育解除柞蚕蛹与对照滞育蛹不同组织中该基因的表达谱;采用实时定量PCR(qPCR)分析其在长光照下滞育解除过程中柞蚕蛹脂肪体中的相对表达量变化。利用3,5-二硝基水杨酸法检测脂肪体中海藻糖酶活力的变化,同时采用蒽酮比色法测定其血淋巴中海藻糖含量。【结果】克隆获得柞蚕3个海藻糖酶基因,分别命名为ApTreh1A,ApTreh1B和ApTreh2(GenBank登录号分别为:KU977455,KU977456和KU977457),开放阅读框(ORF)全长分别为1 797,1 635和1 932 bp,分别编码598,544和643个氨基酸。同源序列比对与系统进化树分析表明,ApTreh1A和ApTreh1B为可溶型海藻糖酶(Treh S),ApTreh2为膜结合型海藻糖酶(Treh M)。半定量RT-PCR检测发现,各组织中ApTreh2比ApTreh1的分布更广且表达量更高。qPCR检测发现,ApTreh1A和ApTreh1B在长光照处理后的柞蚕蛹脂肪体中,21 d时表达量都表现出快速升高[分别是对照组(12L∶12D)的2倍和4.7倍],28 d与35 d时下降,42 d时表达量再次升高;ApTreh2随着滞育的解除表达量逐渐升高,28 d时达到最高(约为对照组的2.7倍),42 d时又出现一个小高峰(约2.3倍),后期逐渐下降。长光照下脂肪体中海藻糖酶活力逐渐升高,21 d时达到最高(约18.5 U),35 d时降到最低(约11.2 U),42 d时其酶活力再次略微升高,之后呈下降趋势,与基因表达变化趋势一致。蛹血淋巴中海藻糖含量在长光照条件下呈现出升高趋势,21 d时达到最高,在整个发育时期的含量比对照组要高。【结论】本研究结果表明柞蚕蛹滞育解除过程中海藻糖酶基因表达的变化与蛹脂肪体中海藻糖酶活性、蛹血淋巴中海藻糖含量的变化趋势呈一致性,提示海藻糖酶基因的表达响应在柞蚕蛹滞育解除中发挥重要作用。     

灭幼脲对粘虫不育作用的机理

通过一系列组织学观察和生物化学测定,研究了灭幼脲在粘虫Mythimna seporata(Walkcr)雌蛾体内及其所产卵内的作用及影响。 灭幼脲(50ppm)经雌蛾连续取食后,卵巢中DNA、RNA的合成及卵巢对血淋巴中蛋白质的吸取均被促进, 卵巢细胞核中DNA、RNA及蛋白质的含量均比正常雌蛾增加,卵巢发育及卵子形成亦被促进,产卵量正常。粘虫生殖系统未受灭幼脲的抑制。胚胎发育阶段细胞核及线粒体中DNA的含量均因雌蛾取食灭幼脲而降低,但胚胎发育并不中断,幼虫分化可达黑头期。灭幼脲导致粘虫所产卵不能孵化的主要原因在于幼虫分化过程中体壁及气管系统的形成受到严重抑制。体壁皮细胞缩小呈扁平形,无细胞核,连成线状,有断裂;原表皮缺乏或极薄。虫体内无气管分布。幼虫死于卵壳内。灭幼脲的不育作用和直接杀卵作用实质上都是对正在分化幼虫中几丁质合成的抑制。本文对灭幼脲的毒理机制进行了讨论。     

葱蝇非滞育、 冬滞育和夏滞育蛹发育和形态特征比较

昆虫非滞育、 冬滞育和夏滞育蛹具有不同的生理和发育过程。本研究以葱蝇Delia antiqua作为模式种, 以黑腹果蝇Drosophila melanogaster蛹的发育形态特征和命名为参照, 用解剖、 拍照、 长度测量和图像处理等方法系统地比较研究了非滞育、 冬滞育和夏滞育蛹的发育历期和形态变化, 重点在头外翻和滞育相关发育和形态特征, 目的在于弄清非滞育、 冬滞育和夏滞育蛹发育和形态特征差异, 为滞育发育阶段的识别、 滞育分子机理研究奠定形态学基础。冬滞育蛹的滞育前期、 滞育期和滞育后期分别为4, 85和27 d, 夏滞育蛹的滞育前期、 滞育期和滞育后期分别为2, 8和22 d。从化蛹至头外翻完成为滞育前期, 头外翻完成约10 h内复眼中央游离脂肪体可见。头外翻的完成是滞育发生的前提, 非滞育、 夏滞育和冬滞育蛹头外翻发生在化蛹后的48, 36和83 h, 在头外翻过程中发育形态没有差异。头外翻的过程为： 首先, 头囊和胸部附肢从胸腔外翻, 头部形态出现; 然后, 腹部肌肉继续收缩, 将血淋巴和脂肪体推进头部及胸部附肢。葱蝇蛹在完成蛹期有效积温约15%时进入冬滞育或夏滞育。在滞育期, 蛹的形态一直停留在复眼中央游离脂肪体可见这一形态时期, 且冬滞育和夏滞育的蛹在形态上没有区别。在体长、 体宽和体重上, 冬滞育蛹最大, 夏滞育蛹次之, 非滞育蛹最小。在滞育后期, 在黄色体出现期间, 非滞育蛹的马氏管清楚可见, 呈绿色, 而滞育蛹的马氏管几乎不可见。本研究为认知昆虫滞育生理、 从发育历期和形态上推断滞育发育进程提供了依据。     

草地螟滞育幼虫的蛋白和核酸含量变化

为了阐明草地螟Loxostege sticticalis 滞育的分子调控基础, 本研究应用Trizol法、 DNA和蛋白质定量试剂盒、 SDS-PAGE电泳技术分别对进入滞育1, 2, 3和4个月、 解除滞育以及非滞育草地螟老熟幼虫中的核酸含量、 总蛋白含量和组分的变化进行了测定。结果表明： 滞育不同月份幼虫的总RNA含量显著低于非滞育幼虫的总RNA含量（P＜0.05）; 解除滞育幼虫的总RNA含量显著高于滞育2, 3和4个月的幼虫, 但仍显著低于非滞育的对照组。滞育不同月份、 非滞育以及解除滞育幼虫的总DNA含量没有显著差异（P＞0.05）。RNA/DNA比值随滞育的开始而显著下降, 随着滞育的结束而显著上升。滞育不同月份的幼虫总蛋白含量显著高于非滞育幼虫的总蛋白含量（P<0.05）, 但解除滞育与非滞育幼虫的总蛋白含量无显著差异。利用SDS-PAGE电泳分析发现滞育幼虫体内存在滞育关联蛋白, 蛋白条带在24 kDa左右, 但非滞育和已经解除滞育的幼虫则没有该蛋白条带。这些结果表明, 总RNA含量的降低、 RNA/DNA比值下降、 总蛋白含量的升高, 以及24 kDa蛋白的存在是草地螟幼虫滞育的主要生理特征。     

柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化

【目的】克隆柞蚕Antheraea pernyi海藻糖合成酶(trehalose-6-phosphate synthase,TPS)基因,并对其进行组织表达分析,探讨该基因在柞蚕滞育蛹解除滞育过程中的表达规律,为阐明柞蚕滞育期间碳水化合物代谢规律与蛹滞育解除的关系提供数据支持。【方法】利用PCR及3'RACE技术从柞蚕幼虫脂肪体组织中克隆得到TPS基因,并进行生物信息学分析;RT-PCR检测该基因在柞蚕幼虫各组织中的表达分布,进一步采用Real-time PCR分析柞蚕滞育蛹解除滞育过程中该基因在脂肪体组织和血淋巴中的表达量变化。【结果】克隆获得柞蚕海藻糖合成酶基因并命名为ApTPS。其开放阅读框长2 487 bp,编码828个氨基酸,蛋白预测分子量为93.19 k D,等电点(p I)4.61;无信号肽,无跨膜区。蛋白质亚细胞定位预测该蛋白定位于细胞质中;蛋白质结构域分析表明,ApTPS有两个保守功能区:TPS(第22-497位氨基酸)和TPP(第532-772位氨基酸)。组织特异性分析表明,ApTPS基因在柞蚕幼虫脂肪体中表达量最高;柞蚕解除滞育过程中,ApTPS在脂肪体和血淋巴中的表达量均有所升高,且显著高于对照组(P0.05),但血淋巴中表达量的升高滞后于脂肪体。【结论】结果提示ApTPS参与了柞蚕蛹滞育中碳水化合物代谢调控并在其中发挥重要作用,与柞蚕蛹滞育解除关系密切。     

山黧豆胚胎发育过程中ODAP和一些大分子物质含量的变化

应用微量分析方法检测了山黧豆胚胎发育过程中ODAP毒素含量和核酸、蛋白质、糖类等大分子物质的含量变化。结果表明:每粒种子的ODAP含量随着胚的发育而增加。每粒种子DNA量随着细胞的迅速分裂而增加,R、蛋白质、淀粉含量随着胚的发育而成倍地增加,当进入心形胚时这些物质的增加更为迅速。如以每克干重中的含量来表示,那么ODAP、DNA及可溶性糖含量则随胚的发育而下降,其它大分子物质含量在胚发育前期升高,进入心形胚时,这些物质达到最高峰;到鱼雷胚时,这些物质含量开始下降,直到胚基本分化完全时,降到最低点;只有酸性蛋白质含量一直保持增长。     

IDENTIFICATION OF A PROTEIN FRACTION IN THE OCCIPITAL CORTEX OF THE MONKEY RAPIDLY LABELLED DURING THE EXPOSURE OF THE ANIMAL TO RHYTHMICALLY FLICKERING LIGHT

Monkeys exposed to a rhythmically flickering light (flicker frequency 7/sec, intensity 1614 lumens/m²) show a higher incorporation of intracisternally administered l -(U-³H)-lysine into proteins of the visual cortex as compared to monkeys kept in darkness. An increase in specific radioactivity is noticed in both the soluble and particulate (including membrane linked) proteins. The 105,000 g supernatant proteins from the visual cortex have been fractionated on DEAE-cellulose columns followed by resolution of each fraction on polyacrylamide gels. The results suggest that there is a group of acidic low molecular weight proteins whose synthesis is significantly stimulated during the exposure of the animal to flickering light. The fractions give immunological cross-reaction with anti S-100 Serum.     

SUBCELLULAR LOCALIZATION OF TRYPTOPHAN-5-MONO-OXYGENASE IN BOVINE PINEAL GLANDS AND RAPHE NUCLEI

Abstract— Tryptophan-5-mono-oxygenase from both bovine raphe nuclei and pineal glands was activated by preincubation with dithiothreitol and ferrous ion at pH 8.5. The optimum pH for the enzyme activity lies between pH 6.4 and 7.3. Preincubation increased the activity of the enzyme from raphe nuclei by about 20 times in both the homogenate and 105,000 g precipitate prepared from it. Activity in the 105,000 g supernatant fraction was about trebled. Corresponding increases in pineal gland enzyme activity were noted: 100 times in homogenate and 105,000 g precipitate and 15 times in 105,000 g supernatant fluid. Total recoveries of activated enzyme from the homogenate prepared in hypo-osmotic medium, in the 105,000 g supernatant and precipitate, were 87.1% and 79.0% for raphe nuclei and pineal glands respectively. Of this, 89.5-91.3% in the case of the raphe nuclei and 76.0-82.0% in the case of the pineal glands, was found in the precipitate. In contrast, 85-90% of the lactate dehydrogenase activity was found in the supernatant fraction. The results of subcellular fractionation revealed that the raphe nuclear enzyme was located in both 'mitochondrial' and 'microsomal' fraction while the pineal gland enzyme was effectively restricted to the 'mitochondrial' fraction. The structural characteristics of the fraction were confirmed by electron microscopy.     

cAMP-dependent protein kinase in sea urchin embryos

cAMP-dependent protein kinase in the supernatant fraction of the homogenate of sea urchin eggs and embryos obtained by centrifugation at 105,000g was investigated in the present study. In the previous report, the dissociation constant between cAMP-binding proteins and cAMP changed during the development. This suggests that the nature of cAMP-dependent protein kinase, which has been well established to be the major cAMP receptor, changes during the development. In the present study, four protein kinases were separated through DEAE-cellulose column from the supernatant of unfertilized egg homogenate. One of them was cAMP-dependent protein kinase. The others were cAMP-independent ones. One among them was phosvitin kinase, and the others were not identified at present. The activity of cAMP-dependent protein kinase gradually increased during a period from fertilization to the swimming blastula stage. During this period, cleavages occurred at a high rate, and the rate decreased after hatching out. Thus, it is supposed that cAMP-dependent protein kinase in the supernatant may take a part in the mechanism of cleavage. The activity, however, became very low at the mesenchyme blastula, the gastrula, and the pluteus stages. cAMP-binding capacity was observed in the sedimentable fraction and the supernatant fraction, respectively, obtained by 105,000g centrifugation at all stages examined. If the structure-bound cAMP-binding protein is also cAMP-dependent protein kinase, it may play different roles in the mechanism of development.     

PROPERTIES AND METABOLISM OF SOLUBLE LIPOPROTEINS CONTAINING CHOLINE AND ETHANOLAMINE PHOSPHOLIPIDS IN RAT BRAIN

Abstract— Choline and ethanolamine phospholipids in the 105,000 g supernatant fraction of rat brain exhibited density and electrophoretic properties consistent with their binding to protein. About 40% of these two phospholipids were bound to soluble lipoproteins, whereas the remainder appeared to be associated with particulate complexes. Following intracranial injection of [2-³H]glycerol, the specific radioactivities of the choline and ethanolamine phospholipids in the supernatant fraction were higher than those in the microsomal fraction at all time points examined, from 15 min to 12 h after injection. The properties of cytoplasmic lipoproteins containing choline and ethanolamine phospholipids have been compared with those which we have previously described containing sulphatide.     

烟草愈伤组织继代培养与分化期间核酸代谢的比较研究

柳叶烟草愈伤组织在分化和芽原基形成期间,DNA 和RNA 含量均高于继代培养物;在芽原基形成后和幼芽生长期间(12天以后),DNA和RNA 含量持续上升,而同期继代培养物巳进入生长静止期,DNA 和RNA 含量基本不变或略有下降。根据RNA 电泳结果还进一步分析了两种愈伤组织培养物各RNA 组分变化与总RNA 含量变化的关系。分化培养物在芽原基形成时有明显升高的RNase 活性峰和持续上升的RNA 合成速率;而此时期继代培养物的RNase 活性及RNA 合成能力均较低;分化愈伤组织的DNA 合成速率在幼芽生长期间仍维持上升趋势,且显著高于同期继代愈伤组织的合成速率。这些结果表明,烟草愈伤组织分化培养物比继代培养物有更旺盛的核酸代谢能力。     

Ca2+-dependent phosphorylation of rat ovary proteins

A 105,000 × g supernatant fraction from prepubertal rat ovaries was incubated in the presence of [γ-³²P]ATP. Phosphorylated proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and identified by autoradiography. Inclusion of Ca²⁺ in the phosphorylation reaction promoted a selective ³²p incorporation into two proteins of M_r = 95,000 and 50,000. Inclusion of chlorpromazine with Ca²⁺ blocked the Ca²⁺-stimulated increase of ³²p incorporation. Our results demonstrate the presence of Ca²⁺-stimulated protein phosphorylation system capable of recognizing endogenous substrate proteins in the prepubertal rat ovary.     

平贝母体细胞胚胎发生中的核酸与蛋白质合成研究

在获得比较理想的平贝母体细胞胚胎发生体系的基础上,我们应用放射性同位素液体闪烁计数技术测定了平贝母体细胞胚胎发生过程中球形胚、心形胚、鱼雷胚、子叶胚和成熟胚等时期的DNA、RNA和蛋白质合成动态,实验表明,从球形胚到子叶胚,核酸与蛋白质的合成速度递增。在子叶胚前期RNA合成达到高峰,在子叶胚期蛋白质合成达到高峰,在子叶胚后期DNA合成达到高峰,核酸与蛋白质合成速度的变化与胚体细胞增殖及器官分化相关联。     

THE ISOLATION AND PARTIAL CHARACTERIZATION OF THE PYRENOID PROTEIN OF EREMOSPHAERA VIRIDIS

The pyrenoids of Eremosphaera viridis, a green alga, were isolated by density gradient centrifugation and their physical and enzymatic properties were studied. The ultraviolet absorption spectrum of sodium dodecyl sulfate (SDS) extracts of pyrenoids showed a single peak at a wavelength of 277 nm, indicating the presence of protein and the probable absence of nucleic acid. Upon electrophoresis on polyacrylamide gels containing SDS, 16 bands were resolved of which two, together, accounted for 90% of the total protein on the gels. The molecular weights of these two proteins were estimated to be 59,000 and 12,300 and the ratio by weight of the larger to the smaller protein was found to be 2:1. The physical and enzymatic properties of these two proteins were found to closely resemble the properties reported in the literature for the subunits of fraction I protein. Both pyrenoids and fraction I protein are localized in the chloroplast, and both have two principal protein components. The molecular weights and relative ratio of the two pyrenoid components are very similar to those of the two components of fraction I protein. The pyrenoid was found to contain a high specific activity of ribulose-1,5-diphosphate carboxylase which is the same enzymatic activity exhibited by fraction I protein. The presence of ribose-5-phosphate isomerase and ribulose-5-phosphate kinase activities was also noted in pyrenoid preparations. It is suggested that the pyrenoid contains fraction I protein and possibly other enzymes of the Calvin-Bassham carbon dioxide fixing pathway.     

Nucleic acids and protein content in relation to growth and regression of buffalo (Bubalus bubalis) corpora lutea

Information on nucleic acids and protein content of buffalo corpus luteum (CL) in relation to growth, development and regression is not available. An experiment was thus conducted to investigate the variation and relationship between nucleic acids and protein content in CL during different developmental stages and to determine the qualitative differences in protein constituents in any of these stages. Buffalo corpora lutea of different developmental stages viz., developing (day 5-10, n = 16), developed (day 11-17, n = 12) and regressed (day 18-21, n = 10) stages were collected from non-pregnant and -pathological genitalia (n = 38). The DNA, RNA and protein content in tissue extracts were determined and the proteins in pooled samples were analyzed by polyacrylamide gel electrophoresis. Developing stage CL had more total and per gram tissue level of DNA and RNA with significant positive relationship with total and per gram RNA and protein contents. Although there was no significant difference in total weight, a significant decrease in total DNA as well as per gram level of DNA and RNA was observed in developed stage compared to developing stage CL. The total protein content in developed stage CL was compared to developing and regressed stage CL. Non-denaturing PAGE analysis of CL proteins of different stages showed five protein bands of 210, 190, 82, 68 and 66 kDa and one that migrated with the dye front in all the stages however, not shown any differences in banding pattern. Denaturing PAGE showed 15 bands viz., 205, 66, 53, 42, 35, 27, 24, 22, 20, 18, 17, 14, 9, 7.5 and 6.5 kDa. Out of these 66 and 53 kDa bands appeared with maximum intensity in all the three stages of CL. Comparison of bands between the three stages revealed five 57, 31, 27, 19 and 16 kDa stage-specific bands in regressed stage CL. The present study indicated that the DNA, RNA and protein content of buffalo CL varied with the stages of development and regressed stage CL contained some unique protein bands which were not observed either in developed or developing stage CL.