DROSOPHILA INFORMATION NEWSLETTER Volume 16, October 1994 The Drosophila Information Newsletter has been established with the hope of providing a timely forum for informal communication among Drosophila workers. The Newsletter will be published quarterly and distributed electronically, free of charge. We will try to strike a balance between maximizing the useful information included and keeping the format short; priority will be given to genetic and technical information. Brevity is essential. If a more lengthy communication is felt to be of value, the material should be summarized and an address made available for interested individuals to request more information. Submitted material will be edited for brevity and arranged into each issue. Research reports, lengthy items that cannot be effectively summarized, and material that requires illustration for clarity should be sent directly to Jim Thompson (THOMPSON@AARDVARK.UCS.UOKNOR.EDU) for publication in DIS. Materials appearing in the Newsletter will be reprinted in DIS. Back issues of DIN are available from FlyBase in the directory flybase/news or in News/ when accessing FlyBase with Gopher. Material appearing in the Newsletter may be cited unless specifically noted otherwise. Material for publication should be submitted by e-mail. Figures and photographs cannot be accepted at present. Send technical notes to Carl Thummel and all other material to Kathy Matthews. The e-mail format does not allow special characters to be included in the text. Both superscripts and subscripts have been enclosed in square brackets; the difference should be obvious by context. Bold face, italics, underlining, etc. cannot be retained. Please keep this in mind when preparing submissions. To maintain the original format when printing DIN, use Courier 10cpi font on a standard 8.5" x 11" page with 1" margins. Drosophila Information Newsletter is a trial effort that will only succeed if a broad segment of the community participates. If you have information that would be useful to your colleagues, please take the time to pass it along. The editors: Carl Thummel Kathy Matthews Dept. of Human Genetics Dept. of Biology Eccles Institute - Bldg. 533 Indiana University University of Utah Bloomington, IN 47405 Salt Lake City, UT 84112 812-855-5782; FAX/2577 801-581-2937; FAX/5374 MATTHEWK@INDIANA.EDU CTHUMMEL@HMBGMAIL.MED.UTAH.EDU MATTHEWK@INDIANA.BITNET *** DIN 16 To add your name to the Newsletter distribution list, send one of the following E-mail messages from the account at which you wish to receive DIN. Via Internet -- To: LISTSERV@IUBVM.UCS.INDIANA.EDU Subject: Message: SUB DIS-L Your real name Via Bitnet -- To: LISTSERV@IUBVM Subject: Message: SUB DIS-L Your real name LISTSERV will extract your user name and node from the E-mail header and add you to the list. Use your Internet address if you have one. You will receive confirmation by E-mail if you have successfully signed on to the list. If you are on the list and do not wish to receive DIN, or you want to remove a soon-to- be-defunct address, replace SUB in the above message with UNS. The SUB command can also be used to correct spelling errors in your real name; the new entry will simply replace the old as long as it was sent from the same USERID@NODE address. *** DIN 16 DIN Vol. 16 TABLE OF CONTENTS >Introduction to Drosophila Information Newsletter >How to subscribe to the Newsletter >TABLE OF CONTENTS >ANNOUNCEMENTS >New Stock Center at Indore, India >Bloomington Stock Center news >Training opportunities at Univ. of Hawaii >REQUESTS FOR MATERIALS >Materials in 99C1-E1 >Deficiencies and P{w[+]} lines >D. simulans with w[+]-marked P inserts >MATERIALS AVAILABLE >Chovnick stocks >TECHNICAL NOTES >An improved devitellinization technique for x-gal staining *** DIN 16 ANNOUNCEMENTS NEW STOCK CENTER AT INDORE, INDIA A new Drosophila Stock Center has been established at Devi Ahilya Vishwavidyalaya, Indore, India; it emphasizes the collection of P insertion, enhancer trap, and other mutant lines. The center is funded by the Indian government's Department of Biotechnology, and is run by Pradip Sinha. Dr. Sinha invites all Drosophila workers to contribute P insertion and other mutant stocks to the collection. The center can be reached by mail at School of Life Sciences, Devi Ahilya Vishwavidyalaya, Vigyan Bhavan, Khandwa Road, Indore-452001, India, or by FAX at 91-731- 473063 (e-mail is not yet available). The Indian Stock Center stock list will be added to FlyBase as soon as a computerized version is available. *** DIN 16 BLOOMINGTON STOCK CENTER NEWS * We will be away for a FlyBase meeting (plus some vacation) October 6 through October 17. Requests received by 11AM on Wednesday, October 5, will be shipped October 10. Requests received between 11AM Oct. 5 and 11AM Oct. 20 will be shipped October 24. * Funding for the stock center has been renewed for the next 5 years, effective September 15, 1994. The collection is now jointly supported by NSF (Biological Instrumentation and Resources) and NIH (the National Center for Research Resources, the National Institute of General Medical Sciences, and the National Eye Institute). The program officers with responsibility for the stock center award are Dr. Machi Dilworth at NSF (mdilwort@nsf.gov) and Dr. Elaine Young at NIH (elainey@ep.ncrr.nih.gov). Thanks to everyone who helped bring this about. Our current funding agreement requires that we institute a cost-recovery program. We will finalize the plans for this program and contact each user group with complete details latter this fall. * The stocks search function on FlyBase has been changed to search all three melanogaster center lists at once. A stock center code precedes each stock number (B for Bloomington, M for Mid-America, and U for Umea) and the center's name and an e-mail address for ordering stocks is included as the last line of each stock record. Please order stocks from the correct center. * The Bloomington stock list on FlyBase is updated whenever new stocks become available. If a stock does not appear on our list we do not have it. *** DIN 16 TRAINING OPPORTUNITIES AT UNIVERSITY OF HAWAII Graduate Research Assistantships (Ph.D. or M.S.). The University of Hawaii announces the availability of graduate assistantships in Ecology, Evolution & Conservation Biology (EECB). Students can study any aspect of ecology, evolution and/or conservation in Hawaii or the Pacific. Much of the Hawaiian biota consists of species swarms which have arisen from single ancestors, and, with the islands arranged in chronological order, provide an unusual opportunity for examining micro- evolutionary events. In addition, the high local endemism, and endangered or threatened status of much of the biota, allows investigation of critical conservation issues. For readers of this newsletter, we draw particular attention to students interested in pursuing the developmental and/or evolutionary genetics of the Hawaiian Drosophila. Such individuals might wish to contact Dr. Terrence W. Lyttle for further information about such research opportunities (tlyttle@uhunix.uhcc.hawaii.edu). Deadline for receipt of all application materials is February 1, 1995. Assistantships will commence in August 1995. For application information, contact Kenneth Kaneshiro (Chair) or Rosemary Gillespie (Associate Chair), Center for Conservation Research and Training, University of Hawaii, 3050 Maile Hawaii, Gilmore 409, Honolulu, HI 96822. (808)956-8884, gillespi@uhuniv.uhcc.hawaii.edu. *** DIN 16 REQUESTS FOR MATERIALS MATERIALS in 99C1-E1 David Bilder, Dept. of Developmental Biology, Stanford Univ. School of Medicine, Stanford, CA 94305-5427, USA. (415)497-2057, bilder@cmgm.stanford.edu. I would appreciate hearing about genetic and molecular information for working in the 99C1-E1 region: P insertions, lethal complementation groups, chromosome aberrations, chromosome walks etc. Thank you. *** DIN 16 DEFICIENCIES AND P{w[+]} LINES Joan E. Wilson, Dept. of Biological Sciences, Gilbert Building, Stanford University, Stanford CA 94305-5020. 415-725-8778, fax/9688, wilsonje@leland.stanford.edu. Looking for deficiencies in 29C-D, 30D-31B, 44C-46C, 48, 91, 92D-93C. Also any P{w[+]} inserts in or around 29-31 or 91-93. *** DIN 16 P{w[+]} SIMULANS STRAINS Dominique Joly, CNRS, Lab. Populations, Genetique et Evolution 91198 Gif sur Yvette Cedex, France. joly@sunbge.bge.cnrs-gif.fr, Fax: 33 1 69 82 37 34. I am studying the genetic basis of sperm length in Drosophila. For that purpose, I would be very interested in any D. simulans strains carrying a P-white[+] element. I am to your disposal for any further indications on the experiments that I would like to realize with those strains. Thanks in advance for your help. *** DIN 16 MATERIALS AVAILABLE Arthur Chovnick, Dept. of Molecular & Cell Biology, U. of Connecticut, Storrs, CT 06269-2131, USA. chounick@uconnvm.uconn.edu. The Chovnick laboratory will not be able to continue to maintain and provide cultures of rosy region stocks beyond January 1, 1995. We will honor requests for these stocks until then. This collection includes rosy wildtype isoalleles with known sequence differences as well as mutations induced on the known isoalleles. Mutant sites cover the entire gene from 5' promoter to the 3' poly T site, and include Ambers, Opals, Frameshifts, electromorphs, transitions, transversions, deletions, duplications and transposable element insertions. They include complimenters, non-complimenters, leaky mutants, nulls and a tissue specific overproducer site located in the large 5' intron, as well as 5' and 3' splice site mutants. The available rosy stocks include the mutants summarized in Lindsley and Zimm, pages 606 through 614. Also, we will be discarding stocks carrying mutations in genes surrounding rosy, and located in 87C, 87D, 87E as well as a set of overlapping deficiencies in this area. See Lindsley and Zimm, pages 399 through 402 and page 873. *** DIN 16 TECHNICAL NOTES AN IMPROVED DEVITELLINIZATION TECHNIQUE WITH A HIGH YIELD OF X-GAL STAINED EMBRYOS A. Singh and M. Kango, Drosophila Stock Center, School of Life Sciences, Khandwa Road Campus, DAVV, INDORE-452001 (M.P) INDIA. indra@cat.ernet.in. X-gal staining (5-Bromo-4-chloro-3-indolyl-beta-D- galactopyranoside) is frequently employed to study the spatio-temporal expression of Drosophila genes. These include enhancer trap studies that involve the insertion of P-lacZ transposons in the vicinity of desired genes (1) and studies involving fusion of the lacZ gene to promoters of developmentally expressed genes (2). X-gal staining is the most popular and convenient method for studying the developmental expression of a lacZ reporter gene (2,3). Devitellinization of embryos, however, is not practiced during X-gal staining since hand peeling of the membrane is time consuming and cumbersome while chemical devitellinization (6) results in diffusion of the stain. Lack of a technique for fast and effective devitellinization of X-gal stained embryos limits the scope of this technique in the study of lacZ expression in embryos. We present here an adaptation of the chemical devitellinization technique (6) which meets these requirements. The protocol involves standard X-gal staining that includes dechorionation of the embryos in bleach (5% Na hypochlorite) and subsequent treatments with 0.7% NaCl and 1% Triton X-100 for 5 min each. Embryos are then fixed in equal volumes of 3.7% formaldehyde in citric phosphate buffer, pH 7.6 (4), and heptane for 15 min. The solution is then drained and the embryos are gently dried to allow traces of heptane to evaporate. This is followed by a rinse in citric phosphate buffer (4) and overnight incubation at 30[o]C in incubation buffer (4) with an increased concentration of Triton X-100 (0.5%) as compared to the usual(0.02%) and a saturating amount (4) of X-gal. Stained embryos are then fixed again with 3.7% formaldehyde, 50 mM EGTA and an equal volume of heptane for 15 min. Fixed embryos are flushed with heptane and after adding 1 ml of 5% TCA (Tricarboxylic acid) stained embryos were shaken for 2 min and then allowed to stand for 5 min. The solution is then removed and the embryos are flushed with methanol and shaken vigorously; the devitellinized embryos will sink down. The duration of this exercise must not last longer than 3-4 min. The solution is then replaced with a mixture of fresh incubation buffer and glycerol (1:1). The embryos will first shrink and then recover their normal shape in 1-2 hrs. The devitellinized X-gal stained embryos are then mounted in a glycero gelatin mountant (4). The time of exposure to methanol should be optimum as prolonged exposure of the embryos to methanol results in flaky appearance of the stain possibly due to precipitation of the proteins. In contrast, shorter periods of exposure give poor yields of devitellinized embryos. Use of a higher concentration of Triton X-100 during staining appeared to be important for retaining the X-gal staining of the devitellinized embryos. This technique overcomes the limitations of the handpeeling technique and also prevents the loss of resolution that results from chemical devitellinization methods due to precipitation of the proteins. This technique provides improved resolution of the X-gal stained embryonic cell types. References: 1. E. Bier et al. (1989) Genes and Dev. 3: 1273- 1287. 2. DiNardo, S. et al. (1988) Nature 332: 604-609. 3. O'Kane, C.J. and Gehring, W.J. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 9123- 9127. 4. Ghysen, A. and O'Kane, C. (1989) Development 105: 35-52. 5. Ashburner, M. (1985) : Drosophila : A laboratory manual. Cold Spring Harbor Laboratory Press. 6. Mitchison, T.J. and Sedat, J. (1983) Dev. Biol. 99: 261-264 *** DIN 16