INTRODUCTION
The pathway from a developmental decision to a morphological
change in a cell or tissue can typically be divided into two sets
of events. The first is a cascade of regulatory processes, such as
cell-cell signals and changes in gene expression. Many of the
molecules involved in these processes can be readily identified
by molecular or genetic screens, because their expression
patterns or mutant phenotypes are tissue-specific. However,
these molecules have no direct impact on cell morphology. The
second set of events includes activation of ‘effector’ molecules
that actually drive cell shape changes. Effectors are often cell
adhesion and cytoskeletal proteins that tend to be generally
expressed and have multiple and/or redundant functions. These
properties can make the genetic analysis of effectors rather
difficult; as a result, effectors of many developmental processes
remain unidentified or poorly characterized.
Drosophila provides not one, but several outstanding model
systems to study the regulation and mechanics of tissue mor-
phogenesis. Adult eyes, wings, ovaries and testes can yield a
wide range of mutant phenotypes because each of these organs
is highly differentiated, but dispensable for viability of the fly.
Many upstream regulators of morphogenesis have been char-
acterized for these and other structures. The connection
between these regulators and their effectors may be elucidated
by a variety of methods including genetic interaction strategies
and ‘reverse’ genetics (disrupting the function of proteins
suspected to be important).
We have performed a reverse genetic analysis of the actin-
based motor protein nonmuscle myosin II. Numerous studies
have shown that the actin cytoskeleton plays a major role in
tissue morphogenesis and can produce specific forces within the
cell in direct response to external signals (aspects of this are
reviewed by von Kalm et al., 1995; Hall, 1994). Of all the actin
modulating proteins that can influence cell shape, the myosins
are uniquely suited to generate contractile forces by sliding
actin filaments past each other or along the plasma membrane.
However, the in vivo composition, organization and regulation
of an actomyosin contractile system are well described only in
the specialized case of muscle cells, where contraction is
powered by sarcomeric myosin II (Harrington and Rodgers,
1984; Rayment et al., 1993). In nonmuscle cells, myosin II is
ubiquitous and has been suspected to drive numerous motile
events (Warrick and Spudich, 1987). However, all eucaryotes
studied express multiple, distinct types of myosin, as well as
numerous other motor proteins, complicating the assignment of
1499
Development 122, 1499-1511 (1996)
Printed in Great Britain © The Company of Biologists Limited 1996
DEV6218
Morphogenesis is characterized by orchestrated changes in
the shape and position of individual cells. Many of these
movements are thought to be powered by motor proteins.
However, in metazoans, it is often difficult to match specific
motors with the movements they drive. The nonmuscle
myosin II heavy chain (MHC) encoded by zipper is required
for cell sheet movements in Drosophila embryos. To
determine if myosin II is required for other processes, we
examined the phenotypes of strong and weak larval lethal
mutations in spaghetti squash (sqh), which encodes the
nonmuscle myosin II regulatory light chain (RLC). sqh
mutants can be rescued to adulthood by daily induction of
a sqh cDNA transgene driven by the hsp70 promoter. By
transiently ceasing induction of the cDNA, we depleted
RLC at specific times during development. When RLC is
transiently depleted in larvae, the resulting adult pheno-
types demonstrate that RLC is required in a stage-specific
fashion for proper development of eye and leg imaginal
discs. When RLC is depleted in adult females, oogenesis is
reversibly disrupted. Without RLC induction, developing
egg chambers display a succession of phenotypes that
demonstrate roles for myosin II in morphogenesis of the
interfollicular stalks, three morphologically and mechanis-
tically distinct types of follicle cell migration, and com-
pletion of nurse cell cytoplasm transport (dumping).
Finally, we show that in sqh mutant tissues, MHC is abnor-
mally localized in punctate structures that do not contain
appreciable amounts of filamentous actin or the myosin
tail-binding protein p127. This suggests that sqh mutant
phenotypes are chiefly caused by sequestration of myosin
into inactive aggregates. These results show that myosin II
is responsible for a surprisingly diverse array of cell shape
changes throughout development.
Key words: nonmuscle myosin, spaghetti squash, imaginal disc,
oogenesis, cell migration, Drosophila, egg chamber
SUMMARY
Drosophila nonmuscle myosin II has multiple essential roles in imaginal disc
and egg chamber morphogenesis
Kevin A. Edwards and Daniel P. Kiehart*
Department of Cell Biology, Duke University Medical Center, Box 3709, Durham, NC 27710, USA
*Author for correspondence (e-mail: Dan_Kiehart@cellbio.duke.edu)