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DIN Vol. 16

Kathy Matthews matthewk at fly.bio.indiana.edu
Fri Sep 30 17:27:17 EST 1994


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 at 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
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     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 at INDIANA.EDU
CTHUMMEL at HMBGMAIL.MED.UTAH.EDU          MATTHEWK at INDIANA.BITNET
***

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***
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 
***

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. 
***

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 at nsf.gov) and Dr. Elaine Young at NIH
(elainey at 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.  
***

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 at 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 at uhuniv.uhcc.hawaii.edu.
***


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 at 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.
***

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 at 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. 
***

P{w[+]} SIMULANS STRAINS
Dominique Joly, CNRS, Lab. Populations, Genetique et Evolution
91198 Gif sur Yvette Cedex, France. joly at 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.
***

                       MATERIALS AVAILABLE

Arthur Chovnick, Dept. of Molecular & Cell Biology, U. of
Connecticut, Storrs, CT 06269-2131, USA.
chounick at 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.
                                     
                         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 at 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
***



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