Differentiation of the melanotrophic cells of rat pituitary primordium in organotypic culture in defined medium

Summary Organotypic cultures, in defined medium, of pituitary primordia obtained from 15-day-old rat fetuses were performed in order to study the in vitro differentiation of melanotrophic cells. The morphological and ultrastructural features of the transplants resembled those of the gland developing in vivo. In situ hybridization on semi-thin sections, using a ³⁵S-labelled oligonucleotide probe, revealed pro-opiomelanocortin-mRNA-containing cells on the first day of culture in the anterior lobe and after 2–3 days in the intermediate lobe. Immunoperoxidase labelling of adjacent sections showed that the same cells reacted with antibodies against -melanocyte-stimulating hormone (MSH), ₃ and adrenocorticotropic hormone in both lobes. The pro-opiomelanocortin-mRNA-containing cells formed progressively conspicuous areas in the intermediate lobe, which was almost uniformly labelled after 6 days. In the anterior lobe, these cells remained scattered in small cell groups, and colloidal gold immunolabelling showed the progressive disappearance of MSH labelling from the secretory vesicles in cells exhibiting morphological features of adult corticotrophic cells. Both the MSH content of the explants and MSH release into the culture medium increased with time. Treatment with the dopamine agonist bromocriptine induced a strong dose-dependent decrease in MSH secretion, which was significant after 3 days in culture, indicating that dopamine D₂ receptors are able to regulate hormonal release of melanotrophic cells at early stages. This system constitutes a suitable model for further studies of factors controlling cell differentiation and cellular interactions involved in histogenesis.     

TGA cysteine codons and intron sequences in conserved and nonconserved positions are found in macronuclear RNA polymerase genes of Euplotes octocarinatus.

The gene sequences of the second largest subunits of RNA polymerases I and II of Euplotes octocarinatus, RPA2 and RPB2, were determined and compared to the respective known sequences of Saccharomyces cerevisiae. The similarity of the derived polypeptide sequences permitted their assignment to the respective polymerases and allowed the comparison of the zinc binding regions. In frame TGA codons were detected, which are likely to encode conserved cysteinyl residues in the putative zinc-finger region of the RPA2 gene. They were also found in other positions in both the RPA2 and RPB2 genes. The RPB2 gene contains a 30 bp intron close to the 5'-end of its coding region. The 5'-ends of the coding regions of all three genes encoding the largest subunits of the three different polymerases were also analyzed. The zinc finger structures again show the use of TGA codons for conserved cysteinyl residues in two of the genes. An N-terminal intron is located in the RPB1 gene at a conserved position as compared to the respective genes of several other eucarya.     

Phytochemical and morphological support for the existence of two species inMonoclea (Hepaticae)

Recognition of two different species in the liverwort genusMonoclea Hook. (monotypic orderMonocleales), viz.M. forsteri Hook. in New Zealand andM. gottschei Lindb. in the New World, is supported by characteristics of the sporophyte, antheridial receptacle and secondary metabolites.M. gottschei produces the greatest variety of flavonoids and the largest amount of bisbibenzyls ever encountered in a liverwort. In contrast,M. forsteri is poor in secondary metabolites. Two allopatric subspecies are recognized inM. gottschei, based on characteristics of the antheridial receptacle: subsp.gottschei in Chile (Valdivian region, Juan Fernandez Is.) and subsp.elongata Gradst. & Mues, subsp. nova, in tropical America. The exclusive occurrence inMonoclea of glucuronide and galacturonide flavone glycosides and the fact that capsule dehiscence may take place before full elongation of the seta are new arguments in support of the placement ofMonocleales in theMarchantiidae. Publication Nr. 43 of the Arbeitskreis Chemie und Biologie der Moose, Universität des Saarlandes, Saarbrücken. This paper is dedicated to DrElla O. Campbell, Massey University, Department of Botany and Zoology, New Zealand on the occasion of her 80th birthday.     

取食雄蜂蛹粉对龟纹瓢虫和异色瓢虫卵黄发生的影响

本文比较了龟纹瓢虫Proylea japonica和异色瓢虫Harmonia axyridis取食蚜虫和取食雄蜂蛹粉时的卵黄发生情况.当取食雄蜂蛹粉时,体内卵黄蛋白出现迟,积累速度慢,产卵前期长.但用保幼激素类似物ZR512点滴处理后则能达到与食蚜对照相当的水平.ZR512对取食雄蜂蛹粉瓢虫的作用显著大于取食岈虫者.进一步的研究表明,ZR512能促进这两种瓢虫取食雄蜂蛹粉,但对成虫的体重没有明显的影响.因此推论,雄蜂蛹粉基本能够满足这两种瓢虫生殖的营养需要,但对其内分泌有一定的影响,使瓢虫处于类似生殖滞育的状态.本文根据不同食物条件对卵黄蛋白发生的影响不同,建议用卵黄蛋白的量作为生理指标,以快速初步筛选和评估人工饲料.     

两种品系油菜植株成分与蚜虫种群消长及成蚜翅型的关系

本文研究两种品系油菜植株成分与桃蚜(Myzus persicae(Sulzer))、萝卜蚜(Lipaphis erysimi(Kaltenbach))的种群消长及成蚜翅型的关系.经分析得出如下结果:1.桃蚜种群消长与苏氨酸、赖氨酸、组氨酸、丙氨酸、硬脂酸等含量有关;桃蚜成蚜无翅率与精氨酸、谷氨酸、酪氨酸、异亮氨酸等含量有关.2.萝卜蚜种群消长与苏氨酸、蛋氨酸、异亮氨酸、苯丙氨酸、酪氨酸、含水量有关;萝卜蚜成蚜无翅率与含水量.亚麻酸.苏氨酸、油酸,天门冬氨酸、丝氯酸、亚油酸等含量有关.     

Variable expression of latent membrane protein in nasopharyngeal carcinoma can be related to methylation status of the Epstein-Barr virus BNLF-1 5''-flanking region.

Seven virus-coded proteins, the nuclear proteins EBNA-1 to EBNA-6 and the latent membrane protein (LMP), are regularly expressed in Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines. In nasopharyngeal carcinoma (NPC), only EBNA-1 is regularly expressed; LMP is detected in about 65% of the tumors. In Burkitt's lymphoma tumors only EBNA-1 is expressed. We have recently shown that the methylation patterns of the EBV genome varied between these cell types. In virally transformed lymphoblastoid cell lines of normal origin, the EBV DNA is completely unmethylated. In contrast, in the Burkitt's lymphoma-derived cell line Rael and in a nude mouse-passaged NPC tumor, C15, there was an extensive methylation of CpG pairs. The methylation extended into the coding regions of the two expressed genes, EBNA-1 (in both tumor types) and LMP (in C15). Two presumptive control regions were exempted from this overall methylation: the oriP that contains both an origin of DNA replication and an EBNA-1-dependent enhancer and the 5'-flanking region of the BNLF-1 open reading frame that codes for LMP. The latter was only exempted in the LMP expressing NPC. We have now investigated the relation between expression of LMP and methylation of DNA in the 5'-flanking 1 kb region of BNLF-1, coding for LMP. LMP was methylated in 3 of 12 NPC biopsies that did not express LMP but was partially or totally unmethylated in the remaining 9 that expressed the protein. The three BNLF-1 exons were highly methylated in all the tumors. The oriP region was unmethylated in all the tumors, as in the previously studied Rael cell line and nude mouse-passaged NPC. Also, the BamHI W enhancer region involved in the expression of EBNA nuclear proteins was methylated. None of the biopsies expressed EBNA-2. Our data show that the EBV genomes are highly methylated in NPC tumors. The strong reverse correlation between the methylation of the putative control region of the LMP gene and the expression of LMP suggests that methylation has a role in the regulation of this gene.     

Performance of 133 compounds in the lambda prophage induction endpoint of the Microscreen assay and a comparison with S. typhimurium mutagenicity and rodent carcinogenicity assays.

The Microscreen assay was developed as a means of testing very small samples, as in complex mixture fractionation. It is a multi-endpoint assay which utilizes E. coli WP2s(lambda). Exposure takes place to serial dilutions of the test compound in microtitre wells (250 microliters) followed by sampling from wells in which growth has occurred ('non-toxic wells'). Although a number of different endpoints can be measured, only the prophage induction endpoint (the first one developed) has been extensively tested. Results with 133 compounds are presented. These include 111 compounds which have been tested in the S. typhimurium assay and 66 compounds for which both rodent bioassay and S. typhimurium assay data exists. The concordance for the Microscreen assay and the S. typhimurium assay was 71%. For this group of compounds, the sensitivity of the Microscreen assay in detecting carcinogens was 76% compared with 58% for the S. typhimurium assay. However, the S. typhimurium assay was somewhat more specific (69%) compared with the Microscreen (56%). The overall association between carcinogenicity and Microscreen results was statistically significant (p = 0.029), whereas for the S. typhimurium assay the association with carcinogenicity was non-significant (p = 0.086). The Microscreen assay was able to detect halogenated compounds better than the S. typhimurium assay. The Microscreen assay should prove useful in complex mixture fractionation, or in other situations where sample size is limiting.     

The trk tyrosine protein kinase mediates the mitogenic properties of nerve growth factor and neurotrophin-3.

The product of the trk proto-oncogene encodes a receptor for nerve growth factor (NGF). Here we show that NGF is a powerful mitogen that can induce resting NIH 3T3 cells to enter S phase, grow in semisolid medium, and become morphologically transformed. These mitogenic effects are absolutely dependent on expression of gp140trk receptors, but do not require the presence of the previously described low affinity NGF receptor. gp140trk also serves as a receptor for the related factor neurotrophin-3 (NT-3), but not for brain-derived neurotrophic factor. Both NGF and NT-3 induce the rapid phosphorylation of gp140trk receptors and the transient expression of c-Fos proteins. However, NT-3 appears to elicit more limited mitogenic responses than NGF. These results indicate that the product of the trk proto-oncogene is sufficient to mediate signal transduction processes induced by NGF and NT-3, at least in proliferating cells.     

The trkB tyrosine protein kinase is a receptor for brain-derived neurotrophic factor and neurotrophin-3.

trkB is a tyrosine protein kinase gene highly related to trk, a proto-oncogene that encodes a receptor for nerve growth factor (NGF) and neurotrophin-3 (NT-3). trkB expression is confined to structures of the central and peripheral nervous systems, suggesting it also encodes a receptor for neurotrophic factors. Here we show that brain-derived neurotrophic factor (BDNF) and NT-3, but not NGF, can induce rapid phosphorylation on tyrosine of gp145trkB, one of the receptors encoded by trkB. BDNF and NT-3 can induce DNA synthesis in quiescent NIH 3T3 cells that express gp145trkB. Cotransfection of plasmids encoding gp145trkB and BDNF or NT-3 leads to transformation of recipient NIH 3T3 cells. In these assays, BDNF elicits a response at least two orders of magnitude higher than NT-3. Finally, 125I-NT-3 binds to NIH 3T3 cells expressing gp145trkB; binding can be competed by NT-3 and BDNF but not by NGF. These findings indicate that gp145trkB may function as a neurotrophic receptor for BDNF and NT-3.