1、生物醫(yī)學(xué)世紀(jì)講壇,How to Publish Your Papers in the Top Scientific Journals,魯白 教授,魯白教授,1957年12月生于上海市。1982年獲上海華東師范大學(xué)學(xué)士學(xué)位。1990年獲康乃爾大學(xué)博士學(xué)位后，在洛克菲勒大學(xué)和哥倫比亞大學(xué)從事博士后研究,導(dǎo)師為Paul Greengard和蒲慕明教授。1993年加入羅氏分子生物學(xué)研究所，并任哥倫比亞大學(xué)助理教授。1996年加入NIH兒童健康和人類(lèi)發(fā)育研究所，任突觸發(fā)育和可塑性研究室主任。 主要研究神經(jīng)營(yíng)養(yǎng)因子在突觸發(fā)育和可塑性中的作用。他的研究室是最早提出并發(fā)現(xiàn)神經(jīng)營(yíng)養(yǎng)因子對(duì)神經(jīng)系統(tǒng)突觸傳遞、突觸發(fā)

2、育可塑性有調(diào)控作用的實(shí)驗(yàn)室之一，與幾個(gè)著名實(shí)驗(yàn)室一起開(kāi)創(chuàng)了神經(jīng)營(yíng)養(yǎng)因子的突觸調(diào)控這一新領(lǐng)域。,M. F. Egan, M. Kojima, J. H. Callicott, T. E. Goldberg, B. S. Kolachana, E. Zaistev, A. Bertolino, B. Gold, D. Goldman, M. Dean, B. Lu, (co-corresponding author) and D. R. Weinberger. (2003) The BDNF val66met polymorphism affects activity-dependent secr

3、etion of BDNF and human memory and hippocampal function. Cell 112, 257-269. L. Ma, Y. -Z. Huang, J. Valtschanoff, L. Feng, B. Lu, W. Xiong, R. Weinberg, L. Mei. (2003) Ligand-dependent recruitment of the neuregulin signaling complex into neuronal lipid rafts. J. Neurosci. 23, 3164-3175. J. Wang, C.

4、-Q. Chen, B. Lu, and C. -P. Wu. (2003) GDNF acutely potentiates Ca2+ channels and excitatory synaptic transmission in midbrain dopaminergic neurons. NeuroSignals 12, 78-88. M. Miura, S. Gronthos, M. Zhao, B. Lu, L. W. Fisher, P. G. Robey, and S. Shi (2003) SHED: Stem cells from human exfoliated deci

5、duous teeth. Proc. Natl. Acad. Sci. USA 100, 5807-5812. Y. X. Zhou, M. Zhao, K. Shimazu, K. Sakata, D. Li, C. -X. Deng, B. Lu. (2003) Impairments in cerebellum Purkinje cells and motor function in mice lacking Smad4 in the central nervous system. J. Biol. Chem. 278, 42313 - 42320. J. Du, L. Feng, E.

6、 Zaitsev, H. S. Je, X. Liu, and B. Lu. (2003) Activity- and tyrosine kinase-dependent facilitation of TrkB receptor internalization in hippocampal neurons. J Cell Biol. In press. Z. G. Luo, H. -S. Je, F. Yang, W. C. Xiong, B. Lu, and L. Mei (2003) Activation of geranylgenanyltransferase is essential

7、 for Agrin-induced AChR clustering. Neuron In press. S. X. Bamji, K. Shimazu, N. Kimes, J. Huelsken, W. Birchmeier, B. Lu, L. F. Reichardt. (2003) Regulation of presynaptic assembly and maintenance by -catenin. Neuron In Press. F. Yang, X. He, J. Russell, and B. Lu. (2003) Ca2+ influx independent tr

8、ansmitter release mediated by mitochondrial Na+-Ca2+ exchanger and protein kinase C. J. Cell Biol. In press.,2003 research articles,What Is the Purpose of Doing Research?,It is not about the number of papers It is not about the impact factor of the papers It is not about the Nobel Prize,Publish or p

9、erish 1 Nature = 10 JBC,Basic Elements for Basic Research,Passion,6 Sigma,Silver bullet test,Good Research Is the Key,My English is not good They are biased against Chinese (foreigners),What Is a First-Class Paper/Research?,Major advance in a classic field 干細(xì)胞是如何分化成特定組織細(xì)胞的，膽固醇在人體的正常功用 New techniques

10、 and methods that can be widely used 人類(lèi)基因組研究中的自動(dòng)測(cè)序技術(shù) , PCR, Patch clamp, Ca2+ Imaging, GFP Discoveries with obvious practical implications AIDS virus receptor 的發(fā)現(xiàn), 老年癡呆癥基因的發(fā)現(xiàn) Conceptual breakthrough, novel ideas 神經(jīng)營(yíng)養(yǎng)因子可以促進(jìn)學(xué)習(xí)記憶, RNA干擾現(xiàn)象 Challenge to traditional views, break dogma 腦內(nèi)有可分裂的神經(jīng)干細(xì)胞，打破了傳統(tǒng)觀念

11、 Opening up new area, cross board “細(xì)胞凋亡”現(xiàn)象的發(fā)現(xiàn), 開(kāi)辟了新的科研領(lǐng)域,What Is a Mediocre Paper/Research?,Horizontal growth I made the discovery in rats, you find the same in cat. Filling gaps EGF activates JNK which is known to induce c-Jun expression. You show that EFG enhances c-Jun expression. Working out det

12、ails I found NO induces the production of cGMP, you work out dose response and time course. Support existing idea, “me too” EGF-R endocytosis requires dynamin, PDGF-R too. Follow up CREB binds to CRE. Working out CRE sequence. Incomplete study, preliminary,How to Read Scientific Papers?,The Gilbert

13、way Keep these in mind when you read What is the major question addressed in this paper? Is this question important and why? What are the approaches used in this paper, and whether they are adequate for the questions? What are the novel idea or using innovative approaches? What is the concept coming

14、 out of this paper? Do the results presented support this new concept? Weekly reading of CNS titles Critical, appreciative,What Makes Good Science?,Important and significant Original and innovative Solid and rigorous Unique and unusual,Novelty is essential,語(yǔ)不驚人誓不休,The evaluation process,Editorial st

15、aff Board of Reviewing Editors,20-30%,REJECT,REVIEW,REJECT,70%,ACCEPT (10%),70%,20%,6%,4%,Should your paper go to CNS?,Is it your best ever? Will it have a big impact? Does it interest scientists in other fields? Does it overturn conventional wisdom? Work that represents a large step forward solutio

16、n to long-standing problem different way of thinking broad implications,What helps:,Convincing data Appropriate controls Careful presentation Consideration of all viable alternatives,What doesnt help,The minimal publishable unit. Excessive or unfounded speculation Repeat examples of a known phenomen

17、on Insufficient advance over previously published work,Editorial Policies of Different Journals,Cell/Neuron/Immunity Editorial board does a lot of reviews. Editors discuss and decide Nature sister journals Editors discuss and decide Science Space meeting, board of review editors PNAS Communicate, co

18、ntribute, Track C Others,Procedures for High Profile Journals,Pre-submission inquiry Submit/cover letter Initial screen Send out for reviews Reject/soft reject/revise Rebuttal Revise again Accept,significance/importance general interests unusual/surprise,Initial screening,suggest reviewers, may take

19、 one friends may not always support you “not to review” always honored “soft” and “harsh” reviewers,Selection of reviewers,You,Editors,Cover Letters,main findings significance suggested reviewers “not to review” list who have read,Dear Editor, We would like to submit the enclosed manuscript entitled

20、 GDNF Acutely Modulates Neuronal Excitability and A-type Potassium Channels in Midbrain Dopaminergic Neurons, which we wish to be considered for publication in Nature Neuroscience. GDNF has long been thought to be a potent neurotrophic factor for the survival of midbrain dopaminergic neurons, which

21、are degenerated in Parkinsons disease. In this paper, we report an unexpected, acute effect of GDNF on A-type potassium channels, leading to a potentiation of neuronal excitability, in the dopaminergic neurons in culture as well as in adult brain slices. Further, we show that GDNF regulates the K+ c

22、hannels through a mechanism that involves activation of MAP kinase. Thus, this study has revealed, for the first time, an acute modulation of ion channels by GDNF. Our findings challenge the classic view of GDNF as a long-term survival factor for midbrain dopaminergic neurons, and suggest that the n

23、ormal function of GDNF is to regulate neuronal excitability, and consequently dopamine release. These results may also have implications in the treatment of Parkinsons disease. Due to a direct competition and conflict of interest, we request that Drs. XXX of Harvard Univ., and YY of Yale Univ. not b

24、e considered as reviewers. With thanks for your consideration, I am Sincerely yours,Dear Editor, We would like to submit the enclosed manuscript entitled Ca2+-binding protein frequenin mediates GDNF-induced potentiation of Ca2+ channels and transmitter release, which we wish to be considered for pub

25、lication in Neuron. We believe that two aspects of this manuscript will make it interesting to general readers of Neuron. First, we report that GDNF has a long-term regulatory effect on neurotransmitter release at the neuromuscular synapses. This provides the first physiological evidence for a role

26、of this new family of neurotrophic factors in functional synaptic transmission. Second, we show that the GDNF effect is mediated by enhancing the expression of the Ca2+-binding protein frequenin. Further, GDNF and frequenin facilitate synaptic transmission by enhancing Ca2+ channel activity, leading

27、 to an enhancement of Ca2+ influx. Thus, this study has identified, for the first time, a molecular target that mediates the long-term, synaptic action of a neurotrophic factor. Our findings may also have general implications in the cell biology of neurotransmitter release.,Dear Editor: Enclosed are

28、 copies of a manuscript entitled BDNF and NT-4/5 Promote the Development of Long-Term Potentiation in the Hippocampus, which we wish to be considered for publication in Nature. As you know, there is a great deal of interest and excitement recently in understanding the role of neurotrophins in synaps

29、e development and plasticity. Our manuscript provides, for the first time, the physiological evidence that neurotrophins regulate long-term potentiation (LTP). The main point of the paper is that the neurotrophins BDNF and NT-4 induce an earlier appearance of LTP in developing hippocampus. In contra

30、st to recent Science article by XXs group, we (and several other LTP groups) did not see that BDNF enhance basal synaptic transmission in adullt hippocampus. However, we found that in adult hippocampus, inhibition of BDNF/TrkB activity attenuated LTP, and weak tetanus that normally cannot induce LTP

31、 produced enduring LTP. These findings may have implications in the basic mechanism for regulation of synapse development and long-term modulation of synaptic efficacy. Because of the rather competitive nature of the field and the important implication of our findings, we have not yet presented this

32、 work in any public forum. However, confidential discussion with several prominent neuroscientists such as 111 and 222 have generated tremendous excitement. Thus, we feel that this work is of general interest and is suitable for publication in Nature. We would like to suggest Drs. aaa of Yale Univ.,

33、 bbb of Harvard Medical School, and ccc of Univ. of California-Berkeley, as reviewers for this manuscript. Due to a direct competition and conflict of interest, we request that Dr. XX and YY. not be considered as reviewers. Thank you very much for your consideration.,Titles,Important/significant Une

34、xpected/unusual Function Mechanisms Simple Straight forward Specific,Structure, mechanism, an regulation of the Neurospora plasma membrane H+,Modulation of postendocytic sorting of G-protein-coupled receptors,Distinct molecular mechanism for initiating TRAF6 signaling,Identification of; Role of; Inv

35、olvement of,Sequence of writing,Abstract Figure layout Figure legend Material and methods Results Introduction Discussion,Abstract,Rationale “remain unknown”; “To determine” Summary statement “Here we show” Body Dont go into details; dont use many special terms Significance Must point out, but dont

36、claim too much,Formation of the normal mammalian cerebral cortex requires the migration of GABAergic inhibitory interneurons from an extracortical origin, the lateral ganglionic eminence (LGE). Mechanisms guiding the migratory direction of these neurons, or other neurons in the neocortex, are not we

37、ll understood. We have used an explant assay to study GABAergic neuronal migration and found that the ventricular zone (VZ) of the LGE is repulsive to GABAergic neurons. Furthermore, the secreted protein Slit is a chemorepellent guiding the migratory direction of GABAergic neurons, and blockade of e

38、ndogenous Slit signaling inhibits the repulsive activity in the VZ. These results have revealed a cellular source of guidance for GABAergic neurons, demonstrated a molecular cue important for cortical development, and suggested a guidance mechanism for the migration of extracortical neurons into the

39、 neocortex.,It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material. - J. D. Watson and F. H. C. Crick,Neuronal responses to brain-derived neurotrophic factor (BDNF) are initiated by the activation of the r

40、eceptor TrkB tyrosine kinase (1). In this study we examined whether cholesterol- and glycolipid-rich microdomains, lipid rafts, provide a functional platform for BDNF-dependent signal transduction (2). Using primary culture of cortical neurons, we demonstrated that TrkB was dramatically translocated

41、 into lipid rafts in BDNF-dependent manner (3). This translocation was blocked by the pharmacological effect of general Trk inhibitors, indicating that TrkB activation is required for the translocation mechanism. We also showed that BDNF and TrkB-FL were both concentrated in lipid rafts during devel

42、opment of cerebral cortex, concomitant with that of synaptic vesicle proteins, including soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins and synaptophysin (4). This result, together with the findings that BDNF stimulation caused translocation of synaptophysin i

43、nto lipid rafts (5) and that BDNF-enhanced glutamate release and exocytosis were both attenuated by depletion of cholesterol from the cell surface with methyl-beta-cyclodextrin (MCD), indicates that lipid rafts are essential for BDNF regulation of neurotransmitter release (6).,Brain-derived neurotro

44、phic factor (BDNF) plays an important role in synaptic plasticity but the underlying signaling mechanisms remain unknown. Here we show that BDNF rapidly recruits full-length TrkB (TrkB-FL) receptor into cholesterol-rich lipid rafts from non-raft regions of neuronal plasma membranes. Truncated TrkB l

45、acking the intracellular kinase domain was not translocated, and the translocation of TrkB-FL was blocked by Trk inhibitors, suggesting a role for TrkB tyrosine kinase in the translocation. Disruption of lipid rafts by depleting cholesterol from the cell surface blocked BDNF-dependent TrkB transloca

46、tion. Disruption of rafts also prevented the potentiating effect of BDNF on transmitter release in cultured neurons, as well as that on synaptic response to tetanus in hippocampal slices. In contrast, lipid rafts are not required for BDNF regulation of neuronal survival. Thus, ligand-induced TrkB tr

47、anslocation into lipid rafts may represent a selective signaling mechanism for synaptic modulation by BDNF in the CNS.,A calcium-independent but voltage-dependent secretion (CIVDS) coexists with the calcium dependent exocytosis in dorsal root ganglion (DRG) neurons (1). Here we have investigated the

48、 CIVDS-coupled endocytosis (2). Using optical and membrane capacitance measurements, we show that, in calcium-free medium, either step depolarization or a train of action-potential-like stimulation induce a novel form of rapid endocytosis, which occurs immediately after the CIVDS. Surprisingly, this

49、 calcium-independent endocytosis is strongly dependent on the stimulation frequency (3). H7 suppress the endocytosis, while PKA agonists enhance it (4). Biochemical experiments show that membrane depolarization directly up-regulate PKA in DRG neurons. Our experiments also showed that the frequency d

50、ependency of CIVDS-RE is dynamin-independent (5). Thus, our data indicate that neuronal activity modulates a rapid endocytosis via a Ca2+- and dynamin-independent phosphorylation-dependent manner in DRG neurons (6).,Synaptic vesicle endocytosis is believed to require Ca2+ and the GTPase dynamin. Her

51、e we report a novel form of rapid endocytosis (RE) that is independent of Ca2+ and dynamin in dorsal root ganglion (DRG) neurons. Using FM dye labeling and membrane capacitance measurements, we show that both step depolarization and repetitive stimulation induce RE in Ca2+-free medium. RE also occur

52、s in the presence of a Ca2+ chelator (BAPTA). Inhibition of dynamin function by three different approaches does not affect RE. Protein kinase A (PKA) inhibitors suppress the endocytosis, while PKA activators enhance it. Biochemical experiments demonstrate that membrane depolarization directly up-reg

53、ulated PKA activity. These results reveal a Ca2+- and dynamin-independent form of endocytosis that is controlled by neuronal activity and PKA-dependent phosphorylation in DRG neurons.,Introduction,What do we know about the subjects? Only relevant information should be provided; dont write a review W

54、hat we dont know Rationale Why you want to do it? Dont repeat abstract Approaches How you are going to do it. Significance Make an appeal to general readers,In this study we have examined the role of chromogranins CGA and CGB, in dense-core secretory granule biogenesis. We analyzed the effect of spe

55、cific depletion of either CGA or CGB, using an antisense RNA strategy, on dense-core secretory granule formation in rat pheochromocytoma (PC12) cells, a model neuroendocrine cell line. We also expressed CGA in a pituitary cell line (6T3) lacking the regulated secretory pathway and nonendocrine fibro

56、blast cells to determine its effect on induction of dense-core secretory granule biogenesis and regulated secretion. Finally, we determined whether CGA could regulate the level of other secretory granule proteins in neuroendocrine and endocrine cells, PC12 and 6T3. These studies identified CGA as a

57、key regulator of dense-core secretory granule biogenesis and storage of other granule proteins in endocrine cells.,Results,Logic Need to explain the rationales in the beginning Connections between paragraphs Dont jump,Previous studies have shown that membrane depolarization-triggered Ca2+ influx thr

58、ough L-type VSCCs induces an increase in BDNF mRNA expression in cultured neurons (Zafra et al., 1990 ; Ghosh et al., 1994 ). This increase in BDNF mRNA could be the result of increased transcription initiation, or increased BDNF mRNA stability, or both. To determine if membrane depolarization stimu

59、lates BDNF transcription, we.,Given the finding that Ca2+ influx through L-type VSCCs induces BDNF transcription, experiments were carried out to determine which of the four BDNF promoters is capable of mediating a Ca2+ response. As described above, the rat BDNF gene consists of four distinct 5 exons each driven by a specific promoter and each spliced to a common 3 exon that encodes the BDNF protein. Since each of the four primary BDNF transcripts can be polyadenylated at one of two sites, a total of eight BDNF transcripts are generated. In principle, the eight tran