Differentiation of Extracutaneous Melanocytes in Embryos of the Turtle,Trionyx sinensis japonicus

The present study investigates the mode of differentiation of neural crest-derived melanocytes in the embryos of the soft-shell turtle, Trionyx sinensis japonicus. DOPA reaction-positive melanoblasts were first detected in 10-day-old embryos. Melanocyte differentiation in terms of pigmentation takes place from the day 16 of development. Melanin pigments were found in the dorsal integument as well as in various extracutaneous tissues such as skeletal muscle, dorsal aorta, peritoneum, blood vessels, choroid, lung, bone marrow, fat tissues and in the connective tissue of the nose. These results suggest the presence of a specific environmental regulation of the melanoblast differentiation in the soft-shell turtle.     

芸苔属脱外壁花粉的分离与人工萌发

芸苔属青菜（Ｂｒａｓｓｉｃａ ｃｈｉｎｅｎｓｉｓ）与紫菜苔（Ｂ． ｃａｍ ｐｅｓｔｒｉｓｖａｒ． ｐｕｒｐｕｒｅａ）的花粉经低温水合、热激、渗激三步程序,分离出大量具萌发能力的脱外壁花粉,脱外壁率可高达６０％ 以上。在含有碳源与氮源及Ｒｏｂｅｒｔｓ培养基盐成分的碱性ＰＥＧ 培养基中,首次使芸苔属脱外壁花粉萌发,萌发率可达３３％ ～４１％ 。在扫描电镜下观察了花粉脱外壁与萌发的过程。讨论了不同植物花粉脱外壁的方法与花粉壁生物学特点的对应关系,以及外壁对花粉萌发的可能作用     

懒猴属的核糖体DNA变异及其种间分化关系

用１５种限制性内切酶和人２８Ｓ、１８ＳｒＤＮＡ探针构建了懒猴属各物种核糖体ＤＮＡ重复单位的限制性内切酶图谱。在进化速率较高的非转录间隔区,在大、中、小懒猴中分别定位了２３、２４、２４个酶切位点。大懒猴与中懒猴有１２个位点不同,与小懒猴有１４个位点不同,而中、小懒猴间则只有一个位点的差异。经过计算,大懒猴与中懒猴的遗传距离值为１２．６５％,与小懒猴的差异为１４．２４％,中、小懒猴间的差异则仅为０．７     

云南姬鼠的蛋白多态性及其遗传分化关系

本文采用蛋白电泳技术对来源于云南省若干地区的姬鼠属（Ａｐｏｄｅｍｕｓ）的３种姬鼠──高山姬鼠（Ａ．ｃｈｅｖｒｉｅｒｉ）８只,中华姬鼠（Ａ．ｄｒａｃｏ）３只和大耳姬鼠（Ａ．ｌａｔｒｏｎｕｍ）１只,以及作为外群的同科的绒鼠属的大绒鼠（Ｈａｐａｌｏｍｙｓｄｅｌａｌｏｒｉ）３只进行了分析。共检测遗传座位２７个,发现２１个座位存在多态性。根据蛋白多态的数据对研究对象进行遗传分化关系的探讨,用系统分析软件ＰＨＹＬＩＰ计算它们之间的分化关系,得到了一棵无根系统树。结果表明,作为外群的大绒鼠明显不同于其它３种姬鼠而聚在最外面。８只高山姬鼠个体汇聚成独立的一支,中华姬鼠的３个个体也聚成一支,但大耳姬鼠却聚在中华姬鼠一支中,因此我们认为大耳姬鼠同中华姬鼠的分化时间可能比较晚近。     

Kinetics of retroviral production from the amphotropic ΨCRIP murine producer cell line

Rapidly expanding development and practice of gene therapy requires the availability of large quantities of high titer retroviral supernatants. One way to achieve high retroviral titers is through improved understanding of the kinetics of retroviral production and decay, and the subsequent development of improved cell culture methods. In the present study we investigated the effects of different operational modes on the retroviral production of the NIH 3T3 fibroblast derived amphotropic murine retroviral producing cell line pMFG/CRIP. Semi-continuous culture (exchange of 50% of medium volume daily) was found to promote cell growth and enhance retroviral production. The rapid medium exchange resulted in significantly larger amounts of high titer supernatants and an extended production phase as compared to the batch control cultures. The specific viral productivity of the pMFG/CRIP cells was in the range of 10 to 40 infectious viruses produced per thousand producer cells per day. The CV-1 African Green Monkey kidney cell line was used as the infection target. Lowering the serum level form 20% to 10% improved retroviral production slightly. However, at lower serum levels (1%, 5% and 10% (v/v)) growth of the producer cell line, and thus retroviral production, was directly proportional to the serum level. The half-life of the virus at 37°C was found to be 5.5 hours. Promoting the growth of producer cell lines can improve retroviral vectors titers and viral production. High cell density systems that allow for rapid cell growth and waste product removal are likely to be used to generate high-titer retroviral supernatants.     

Turtle Lung Cells Produce a Melanization-Stimulating Activity That Promotes Melanocytic Differentiation of Avian Neural Crest Cells

We found previously that neural crest cells in turtle embryos migrated into the lung buds and melanocytes were located in the lungs. The finding suggested to us that the lungs provide a stimulatory factor(s) to the differentiation of neural crest cells into melanocytes. We have established lung cell lines to facilitate analysis of the interactions of neural crest cells with the environment in melanocyte development. One cell line, TLC-2, was found to produce a putative melanization-stimulating activity (MSA), which promoted the melanocyte differentiation in vitro of avian neural crest cells. The TLC-2-derived MSA was different from that of basic fibroblast growth factor (bFGF), α-melanocyte stimulating hormone (α-MSH), and steel factor (SLF). Its molecular weight was estimated to be within the range of 150 kD. Our findings suggest that MSA may be a novel factor exercising a positive control over melanocyte differentiation.     

包囊游仆虫休眠包囊的超微结构研究

包囊游仆虫休眠包囊中,各类纤毛器的纤毛基体上方的大部分纤毛杆退化,或仅保留毛基体,有时部分额腹棘毛的毛基体也瓦解消失。残留纤毛的纤毛杆周围微管和中央微管仍具有“9 2”结构特征,也有少数纤毛杆出现2套“9 2”微管共处于一层纤毛膜内的现象。毛基体中周围三联体微管的中央形成微管形结构聚合体,基体附属结构仅存在基体间连接及纤毛器托架的残余物;非纤毛区皮层表膜下未见微管层。纤毛区皮层含纤毛器腔周围微管层(相当于表膜下微管层)、纤毛器深部及附近的微管束和分散的微管群。并且,纤毛区皮层囊泡内含有呈不同形态的纤毛杆结构;大核核孔明显变大,核孔数目减少,核孔内膜附着染色质。     

Induction of Antibiotic Resistance in Paramecium tetraurelia by the Bacterial Gene APH-3'-II

We have generated a transformation marker for Paramecium using a Paramecium expression vector (pPXV) and the open reading frame (ORF) of the bacterial antibiotic resistance gene aminoglycoside 3'-phosphotransferase-II (APH-3'-II or neo^r) from the transposon Tn5. The expression vector contained a small multiple cloning site between the 5' and 3' non-coding regions of the calmodulin gene, and Tetrahymena telomere sequences for the stability of the plasmid in Paramecium. After the neo^r ORF was inserted, the plasmid was referred to as pPXV-NEO. Delivery of approximately 10–20 picoliters of linearized PXV-NEO at > 2000 copies/pl into the macronucleus effected 100% transformation. Southern and Northern blot hybridization showed the presence of neo^r-specific DNA and RNA, respectively, in all of the transformed clones but not in the untransformed clones. The degree of resistance to G-418, and the concentrations of neo^r-specific DNA and neo^r-specific RNA in the clones were proportional to the concentration of the vector injected. We have demonstrated that when the linearized plasmid was injected into the macronucleus, the prokaryotic sequence conferred an antibiotic resistance to Paramecium despite codon-usage differences.     

c-fos expression in the amygdala:In vivo antisense modulation and role in anxiety

Summary 1. The amygdaloid complex is a key structure in mechanisms of fear and anxiety. Expression of the immediate-early gene c-fos has been reported in the central nucleus of the amygdala following various stressors, but the functional role of this phenomenon has remained unknown.2. c-fos expression was observed in the central nucleus when rats were subjected to a pharmacologically validated animal model of anxiety, the Vogel conflict test, but not after mere exposure to the test apparatus. Bilateral amygdala injection of a 15-mer phosphorothioate c-fos antisense oligodeoxynucleotide prior to testing blocked conflict-induced c-fos expression and had behavioral effects similar to those of established antianxiety drugs.3. Separate experiments determined that antisense treatment did not affect conflict behavior by acting on shock thresholds or drinking motivation.4. These findings provide evidence that neuronal activation and c-fos induction in the amygdala may be of importance for mechanisms of fear and anxiety.