DEVELOPMENTAL DYNAMICS 218:80 –93 (2000)
REVIEWS
A PEER REVIEWED FORUM
Integration of Epithelial Patterning and Morphogenesis in
Drosophila Ovarian Follicle Cells
LEONARD L. DOBENS AND LAUREL A. RAFTERY*
Cutaneous Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Charlestown,
Massachusetts
ABSTRACT Drosophila oogenesis involves
the coordinated development of germ cells and
an overlying follicular epithelium. The follicle
cells provide a genetically tractable system to
investigate the cell biology of patterning and
morphogenesis. Follicle cells initially form a
cuboidal epithelium surrounding a syncytium of
nurse cells and oocyte. Epithelial structure is
maintained as these cells reorganize to create
the three dimensional architecture of the egg-
shell. Both long-range and short-range cell-cell
communications pattern the domains of follicle
cells that will create specific eggshell structures.
After terminal differentiation to deposit the egg-
shell proteins, the follicle cells die. This review
summarizes recent progress in understanding
the cell-cell communication that orchestrates fol-
licle cell patterning and migrations. DE-cad-
herin-mediated adhesion is important at several
steps in egg chamber formation and follicle cell
migration. Notch signaling is critical during each
successive round of patterning and migration.
Integration of bone morphogenetic protein
(BMP) and epidermal growth factor (EGF) sig-
nals patterns the elaborate structures of the dor-
sal-anterior eggshell. Dev Dyn 2000;218:80 –93.
© 2000 Wiley-Liss, Inc.
Key words: Drosophila melanogaster; follicle cells;
oogenesis; cell migration; pattern
formation
INTRODUCTION
The Drosophila egg is surrounded by an eggshell
with elaborate structures that enhance survival of the
developing embryo. This eggshell is the product of the
somatic follicle cell epithelium, which develops in con-
cert with the oocyte. Oogenesis requires a series of
interactions between the follicle cells and an underly-
ing cyst of germ cells. At the same time, distinct do-
mains of follicle cells (FCs) are patterned by series of
local cell-cell interactions. Within each domain, the
cells undergo coordinated shape changes and move-
ments, followed by differentiation to create a special-
© 2000 WILEY-LISS, INC.
ized structure of the eggshell. An accompanying review
discusses the establishment and maintenance of the
FC epithelium. Here, we focus on patterning of the FC
fates and how it is linked to the cell shape changes and
migrations that create the three-dimensional architec-
ture of the eggshell.
The follicle cells provide a tractable system to inves-
tigate key questions about the coordination of cell fate
and behavior within an intact epithelial sheet. FCs
first organize a simple cuboidal epithelium; epithelial
structure is maintained through migrations that create
a three-dimensional architecture. FC development re-
quires many of the same genes that regulate morpho-
genesis of other organs (Table 1). How are domains of
cell fate patterned by local and regional cell-cell com-
munication? How are boundaries set, so that morpho-
genesis is uniquely orchestrated within each domain?
How is tissue architecture formed, maintained, and
dissolved? Early on, cell contacts were recognized as
important signals in FC morphogenesis. More recently,
critical roles were demonstrated for secreted signals
such as epidermal growth factor (EGF)-related pro-
teins and bone morphogenetic protein (BMP)-related
members of the transforming growth factor-␤ (TGF-␤)
family. Notch signaling has been implicated in all as-
pects of FC morphogenesis. Understanding the succes-
sive interactions of these patterning signals will shed
light on their interactions in morphogenesis of other
tissues and other organisms.
OVERVIEW OF OOGENESIS
Each Drosophila egg develops as a discrete structure
termed an egg chamber (Fig. 1; reviewed in King, 1970;
Spradling, 1993). Egg chambers are formed at the an-
terior of the ovary, and move posteriorly as they de-
velop. Thus, each ovariole contains a linear array of
developmental stages. The follicular epithelium under-
Grant sponsor: American Cancer Society; Grant number: RPG-96-
103-03-DDC; Grant sponsor: National Institutes of Health; Grant
number: GM60501.
*Correspondence to: Laurel A. Raftery, Cutaneous Biology Research
Center, MGH-East 3rd Floor, Building 149 13th Street, Charlestown,
MA 02129. E-mail: laurel.raftery@cbrc2.mgh.harvard.edu
Received 10 January 2000; Accepted 8 February 2000
 REGULATION OF FOLLICLE CELL MORPHOGENESIS 81

TABLE 1. Genes Involved in Follicle Cell Morphogenesis

Gene name Ovarian requirementa Protein function Referenceb

EGFR signaling
Epidermal growth factor
receptor (EGFR, torpedo) S EGF receptor tyrosine kinase (RTK) 1
fs(1)K10 (K10) G Helix-turn-helix motif transcription factor 2
gurken (grk) G TGF␣ ligand 3
ras S Small GTPase, RTK signaling 4
Raf G/S Raf kinase, RTK signaling 5
argos (aos) S Secreted, EGFR antagonist 6
rhomboid (rho) S Transmembrane protein, RTK mediator 7
sprouty (sty) S Membrane protein, EGF antagonist 8
pointed (ptd) S ETS domain transcription factor 9
bunched (bun) S TSC-22 family transcription factor 10
Notch signaling
Notch (N) S Cell surface receptor 11
Delta (Dl) S Cell surface ligand 11
big brain (bib) S MIP family motif, aquaporin, channel 11
brainiac (brn) G Novel, secreted 12
daughterless (da) S bHLH motif transcription factor 13
cut S Homeodomain transcription factor 14
fs(1)Yb G Novel 15
toucan (toc) G Novel, coiled-coil motif 16
neuralized (neu) S Zinc finger transcription factor 11
egghead (egh) G Novel, secreted 17
BMP signaling
decapentaplegic (dpp) S BMP ligand 18
thick veins (tkv) S BMP receptor ser-thr kinase 18
saxophone (sax) S,G BMP receptor ser-thr kinase 18
Mothers against dpp (Mad) S Smad transcription factor 19
D-mef2 Myogenic transcription factor 20
Hedgehog signaling
hedgehog (hh) S Ligand 21
costal-2 (cos-2) S Kinesin domain, antagonist 22
cubitus interruptus (ci) S Zinc finger transcription factor 23
patched (ptc) S 7-pass transmembrane protein, antagonist 23
JUN kinase signaling
hemipterous (hep) S JUN kinase 39
puckered (puc) S JUN phosphatase 40
Wnt signaling
wingless (wg) S Wnt family ligand 23
Cytoskeletal organization
zipper (zip) S,G Myosin II heavy chain 25
spaghetti squash (sqh) S Myosin regulatory light chain 26
rap1, Dcdc42, Drac1, DRhoL S/G Small GTPase family members 27, 28
␣-spectrin S ␣-spectrin, membrane cytoskeleton 29
karst S ␤-H-spectrin, membrane cytoskeleton 30
Cell adhesion
Fasciclin III (FasIII) S Septate junctions; transmembrane 11
discs large (dlg) S,G Septate junctions, MAGUK family 31
shotgun (shg) S,G E-cadherin, cell adhesion 32
armadillo (arm) S ␤-catenin 24
Other genes and markers
stall S Unknown 33
slow border cells (slbo) S bZIP transcription factor 34
breathless (btl) S FGF receptor tyrosine kinase 35
93F S Stalk cell marker 33
fs(1)cup G Novel; coiled-coil motif membrane protein 36
bullwinkle (bwk) G HMG protein 37
dicephalic (dic) G Unknown 38
A359 S Centripetally migrating FC marker, unknown 19
a
Abbreviations: (G) Germ line, (S) soma.
b
All genes are described in FlyBase (http://flybase.bio.indiana.edu/) Representative references: 1. Price et al. (1989); 2. Cheung
et al. (1992); 3. Roth and Schüpbach (1994); 4. Lee and Montell (1997); 5. Brand and Perrimon (1994); 6. Wasserman and
Freeman (1998); 7. Ruohola-Baker et al. (1993); 8. Reich et al. (1999); 9. Morimota et al. (1996); 10. Dobens et al. (1997); 11.
Ruohola et al. (1991); 12. Goode et al. (1996a); 13. Cummings and Cronmiller (1994); 14. Jackson and Blochlinger (1997); 15.
King and Lin (1999); 16. Grammont et al. (1997); 17. Goode et al. (1996b); 18. Twombly et al. (1996); 19. Dobens et al. (2000);
20. Mantrova et al. (1999); 21. Forbes et al. (1996a); 22. Liu and Montell (1999); 23. Forbes et al. (1996b); 24. Peifer et al. (1993);
25. Young et al. (1993); 26. Edwards and Kiehart (1996); 27. Asha et al. (1999); 28. Murphy and Montell (1996); 29. Lee et al.
(1997); 30. Zarnescu and Thomas (1999); 31. Goode and Perrimon (1997); 32. Niewiadomska et al. (1999); 33. Tworoger et al.
(1999); 34. Montell et al. (1992); 35. Murphy et al. (1995); 36. Keyes and Spradling (1997); 37. Rittenhouse and Berg (1995);
38. González-Reyes et al. (1997); 39. S. Noselli, personal communication; 40. LLD and LAR, in preparation.
82 DOBENS AND RAFTERY
Fig. 1. Follicle cell migrations in the germarium. Anterior is to the left
in these and all subsequent figures. Germ cells divide in region 1 to form
a 16-cell cyst, which flattens as it exits region 2A. Under the influence of
the terminal filament, FCs (depicted in gray) proliferate in region 2B,
where they migrate to contact the posterior side of the flattened cyst. The
cyst is enveloped by FCs to form a stage 1 egg chamber as it leaves
region 2B. FCs that do not contact the germ cyst interleave to form stalks
that connect to adjacent egg chambers at the polar cells.
goes many changes after the egg chamber is formed.
Terminally differentiated FCs die after depositing the
eggshell. The structural details of the eggshell reveal
the organization of the final FC epithelium.
Egg Chamber Formation
The egg chamber forms from two distinct cell types:
the somatic follicle cells, derived from the gonadal me-
soderm, and the germ cells. Egg chambers are assem-
bled at the anterior tip of each ovariole, in a germarium
(Fig. 1). Germline stem cells are maintained at the
distal tip (Lin and Spradling, 1993). Egg chambers
assemble in an adjacent tube of somatic sheath cells,
which is divided into three regions. In region 1, the
pro-oocyte arises from an asymmetric stem cell divi-
sion, and undergoes four incomplete cell divisions to
form a syncytial cyst. As the cyst moves posteriorly
through region 2a, one germ cell is selected to become
the oocyte. The remaining 15 germ cells become nurse
cells. The cyst flattens and moves to region 2b, where a
few FC stem cells reside (Margolis and Spradling,
1995). FCs migrate to encase the cyst in a cuboidal
epithelium.
As the egg chamber buds off from the germarium,
five to eight FCs become stalk cells (Fig. 1). These cells
form stacks that separate adjacent egg chambers. Stalk
cells do not contact the germ cells, and are not part of
the epithelium. Within the epithelium, the polar cells
become distinct from other FCs (Margolis and Sprad-
ling, 1995). These cells are important for subsequent
patterning of the egg chamber.
Morphogenesis of Eggshell Architecture
After leaving the germarium, the egg chamber grows
in size. The germ cells enlarge from synthesis in the
nurse cells and uptake of yolk produced in the fat body
(reviewed in Spradling, 1993). With the exception of
the polar cells, FC proliferation continues through
stage 6 (Orr-Weaver, 1991; Calvi et al., 1998). During
stages 7 to 10, FCs become polyploid through en-
doreduplication. Specific amplification of the chorion
genes occurs after stage 10B.
The architecture of the eggshell is laid out through a
stereotypic sequence of migrations. During stage 9,
most FCs shift posteriorly to form a columnar epithe-
lium over the oocyte (Fig. 2A–C). During this process,
about 30 anterior FCs flatten to form a squamous epi-
thelium over the nurse cells. Simultaneously, 6 –10
anterior FCs, the border cells, migrate between the
nurse cells to the oocyte. The anterior squamous and
posterior columnar FCs maintain a contiguous epithe-
lium over the germline cyst.
Follicle cell behaviors are tightly coordinated with
events in the germ cells during stage 10B. First, the
nurse cells dump their contents into the oocyte, causing
it to swell rapidly. At the same time, FCs at the nurse
cell-oocyte boundary migrate centripetally, to cover the
anterior end of the oocyte (Fig. 2D–F). The squamous
FCs shrink in size, maintaining their association with
the degenerating nurse cells. As the oocyte swells,
mainbody FCs stretch along the anterior/posterior
axis, maintaining the epithelium. FCs surrounding the
oocyte deposit the eggshell. Two dorsal anterior FC
subgroups migrate coordinately to produce the dorsal
appendages of the eggshell. Finally, FCs undergo apo-
ptosis (Chao and Nagoshi, 1999), and are resorbed as
the mature egg exits the ovary.
Structure of the Eggshell
The eggshell (Fig. 3A) is a multi-layered protein
structure deposited by the FCs (Margaritis et al.,
1980). The inner layers include the vitelline mem-
brane, the waxy layer, and the inner chorion layer
(ICL). The outer layers include the endochorion, which
we focus on here, and the exochorion. The proteins that
comprise each layer are deposited by the FCs under
strict temporal control. The roof of the endochorion
shows ridges that correspond to the junctions between
FCs. Thus, FCs leave imprints on the eggshell, so that
the final epithelial organization is evident in the result-
ant egg (Fig. 3B).
The eggshell has regional specializations, both in its
three-dimensional architecture and in the ultrastruc-
ture of the specific layers (Margaritis et al., 1980). The
most prominent features are the dorsal appendages,
long tubes of chorion that project from the anterior-
dorsal surface (Fig. 3C). The chorion of these filaments
is uniquely porous (Fig. 3C inset), suggesting a role in
gas exchange. The anterior-most structure is the oper-
culum, a flat plate ringed on three sides by a prominent
ridge, the collar (Fig. 3A, E). The operculum plate
arises from the centripetally migrating FCs of stage
10B. The micropyle juts out near the ventral collar
(Fig. 3E), and surrounds a canal for sperm entry,
formed by the border cells. The operculum can be fur-
 REGULATION OF FOLLICLE CELL MORPHOGENESIS 83

Fig. 2. Follicle cell rearrangements during mid-oogenesis. Anterior is to the left. ␣-spectrin staining reveals FC shapes (white; ␣-spectrin antiserum
from Pesacreta et al., 1989). A: Stage 8 egg chamber cuboidal epithelium covering nurse cells (NC) and oocyte (O; line marks the anterior edge).
Posterior terminal cells are more rectangular than others. B: Main body epithelium (MBE) changes shape (arrows) and shifts posteriorly (stage 9). C:
Border cells (bc; arrowhead) migrate through the nurse cells; the oocyte increases in size (stage 9). D: Boundary between squamous and columnar
cells (arrow) is the site of presumptive centripetally migrating FCs (CMFC, stage 10A). E: Centripetally migrating FCs further elongate and begin to
cover anterior of oocyte (stage 10B). F: Border cells shift slightly to the dorsal side.

ther subdivided into three regions that form concentric
arcs around the micropyle (Fig. 3E). The collar exhibits
ultrastructural perforations in the chorion and
vitelline membrane and is easily split. Thus, the oper-
culum is an escape hatch for the larva. At the posterior
pole, the aeropyle includes two nested rings of distinct
cell imprints (Fig. 3D, F). FC patterning specifies the
domains that will create each eggshell structure; FC
migrations move these domains to the proper position.
The specialized ultrastructure of the chorion must
arise from different programs of gene expression and
protein secretion as the differentiated FCs deposit the
eggshell.
FOLLICLE CELL MORPHOGENESIS DURING
EGG CHAMBER FORMATION
The formation of the egg chamber involves three
interdependent processes: 1) FC proliferation from
stem cells, 2) FC migration and envelopment of a germ-
line cyst, and 3) FC differentiation to create the stalks
that separate egg chambers. Some defects in these
processes lead to egg chambers with two or more com-
plete germline cysts; in extreme cases, a single epithe-
lial bag full of cysts forms. Other defects can result in
split cysts, in which an egg chamber with too few germ
cells occurs next to one with too many. In either case,
the egg chambers fail to develop, causing reduced fe-
cundity or sterility.
Regulation of Stem Cell Proliferation
A lifetime of egg production depends on continuous
production of FCs from a few stem cells in germarium
region 2b (Fig. 1) (Margolis and Spradling, 1995). Re-
duced FC proliferation results in fused egg chambers
and gaps in the FC epithelium. The somatic and germ-
line stem cells can proliferate independently (Gateff,
1982; McKearin and Spradling, 1990; Margolis and
Spradling, 1995), but may be coordinated (King and
Lin, 1999). FC proliferation in the germarium is regu-
lated by a hedgehog (hh) signal produced by the termi-
nal filament, and requires the signal transduction com-
ponents patched and cubitus interruptus in the FCs
(Forbes et al., 1996a; Forbes et al., 1996b). Hh signal-
ing regulates both FC proliferation and specification of
the stalk/polar cell precursors, suggesting that these
processes are connected.
Follicle Cell-Germline Interactions Mediate
Cyst Envelopment
Assembly of discrete egg chambers requires that FCs
recognize individual germline cysts and migrate be-
tween them. Encapsulation of the germline cyst in-
volves both the envelopment of the cyst and the sepa-
ration of egg chambers by differentiation of stalk cells.
These processes are interdependent, as revealed by
sensitive genetic interactions between several of the
genes involved.
Proliferating FCs migrate between successive cysts,
first contacting the posterior portion of the cyst disc
(Fig. 1) (González-Reyes and St. Johnston, 1998b) and
then surrounding the entire disc. A germ cell to FC
signal promotes initial FC migration and patterning;
this signal either activates or functions coordinately
with Notch signaling. Notch protein levels are tran-
siently elevated in posterior FCs of region 2b, and fused
egg chambers are among the phenotypes engendered
by a temperature-sensitive mutation in Notch (Nts, Xu
et al., 1992). The use of temperature-sensitive muta-
tions has been essential to determine the roles of Notch
signaling, because it functions at multiple times during
84 DOBENS AND RAFTERY

Fig. 3. Specialized features of the Drosophila eggshell. (A–F) Scanning electron micrographs of eggshell chorion. A: Dorso-lateral view of entire
egg. Reference line marks the dorsal midline in this and other panels. B: Dorso-lateral close up of midbody imprints (d ⫽ dorsal; v ⫽ ventro-lateral
imprints). C: Close up views of a dorsal respiratory appendage (da). Inset shows the sharp boundary between specialized exochorion along the ventral
and dorsal sides of the appendage (arrowheads). D: Aeropyle at the posterior pole of the eggshell, with two types of distinct imprints, I and II. E: Close
up view of the anterior eggshell showing the micropyle (mp), dorsal appendages (da) and three regions of distinct cell imprints (I–III). F: Close up view
of posterior imprints shows thin eggshell of region I imprints and waved surface of region II imprints. Scanning electron micrographs are reprinted from
Margaritas et al., 1980, with permission from The Company of Biologists, Ltd.

oogenesis (Xu et al., 1992). Another neurogenic gene,
brainiac (brn), is required in the germline and genet-
ically interacts with Notch (Goode et al., 1996). Notch
interacts with two other genes required in the germ-
line, egghead (egh) and toucan (toc) (Goode et al.,
1996a; Grammont et al., 1997). An alternative hy-
pothesis is that the germline genes are required to
strengthen germ cell-germ cell adhesion, so that FCs
cannot migrate between the germ cells within a wild-
type cyst (Rubsam et al., 1998). However, FCs that
lack the Cut transcription factor can migrate be-
tween germ cells within a cyst (Jackson and Bloch-
linger, 1997), supporting the notion that FCs actively
distinguish the boundary of a cyst. Strikingly, re-
duced Notch levels suppress this phenotype of cut
mutants.
 REGULATION OF FOLLICLE CELL MORPHOGENESIS
The germ cell to FC signal may be DE-cadherin-
mediated adhesion, which is required for several steps
of cyst envelopment (Oda et al., 1993). Encoded by the
gene shotgun, DE-cadherin is an integral membrane
protein that mediates cell-cell adhesion through ho-
mophilic binding. FCs first contact the posterior of the
disc-shaped germline cyst, where the oocyte lies (Fig.
1). In the nascent egg chamber, DE-cadherin levels are
elevated in both the oocyte and the FCs at the anterior
and posterior poles. Expression is required in both cell
types to maintain the oocyte in this posterior position
(Godt and Tepass, 1998; González-Reyes and St.
Johnston, 1998b). When the posterior FCs lack DE-
cadherin, the oocyte associates with DE-cadherin-ex-
pressing anterior FCs. The oocyte position determines
the anterior-posterior polarity of the egg chamber,
which subsequently establishes the anterior-posterior
polarity of the embryo (Godt and Tepass, 1998; Gonzá-
lez-Reyes and Johnston, 1998).
Encapsulation Involves Determination of Stalk
Cells and Polar Cells
Each egg chamber is attached to its neighbors by a
stalk that forms as it leaves the germarium (Fig. 1).
Stalks are essential to maintain separate egg cham-
bers. Fused egg chambers arise from mutations that
cause the early absence of stalk cell precursors (Ruo-
hola et al., 1991; Forbes et al., 1996a) or the later
degeneration of the stalks (Edwards and Kiehart,
1996). The stalks contact the FC epithelium through
two pairs of polar cells, situated at the anterior and
posterior poles of the egg chamber.
A single precursor cell type gives rise to stalk cells
and polar cells (Tworoger et al., 1999). The stalk/polar
precursor is specified by a Hh signal produced from the
terminal filament (Forbes et al., 1996a; Forbes et al.,
1996b). Ectopic Hh expression can induce the forma-
tion of ectopic polar and stalk cells at any position in
the follicular epithelium. The costal2 gene is required
in the FC epithelium to prevent the polar cell fate
(Forbes et al., 1996; Liu and Montell, 1999; Tworoger et
al., 1999).
The level of Notch signal is critical for the proper
subdivision of stalk cells and epithelial FCs. Stalk cell
marker expression is lost when Notch signal levels are
reduced using temperature sensitive mutations in
Notch or its ligand Delta (Ruohola et al., 1991). In
contrast, low level expression of activated Notch in-
duces extended stalk structures (Larkin et al., 1996).
At higher levels of activated Notch, stalk cells fail to
fully differentiate. Both stalk cells and polar cells show
persistent expression of daughterless, which encodes a
transcription factor required for Notch signaling in
several tissues (Cummings and Cronmiller, 1994).
Consistent with this role, daughterless mutations ge-
netically enhance both Notch and Delta mutant encap-
sulation phenotypes. Other genes, such as fs(1)Yb
(Johnson et al., 1995) and cut (Cummings and Cron-
miller, 1994), show genetic interactions with Notch
that implicate them in stalk cell differentiation. Stalk
85
formation requires additional genes, such as rap1 and
stall (Asha et al., 1999; Tworoger et al., 1999), but their
specific roles are unknown. Subtle differences in Notch
and hh phenotypes indicate that these signals regulate
different aspects of stalk formation.
Polar cells are determined in the germarium, where
they cease to divide. Visible differentiation of the polar
cells occurs during stages 6 – 8, when these cells round
up and lose contact with the basal lamina. Polar cells
exhibit unusual subcellular localization of DE-cadherin
and the ␤-catenin Armadillo, and are otherwise dis-
tinct in their subcellular morphology (Ruohola et al.,
1991; Peifer et al., 1993; Oda et al., 1997; González-
Reyes and St. Johnston, 1998b). The polar cells are the
keystone for FC patterning, for they nucleate subse-
quent patterning of the terminal FCs (González-Reyes
and St. Johnston, 1998b; González-Reyes and St.
Johnston, 1998a).
ESTABLISHMENT OF TERMINAL AND
MAINBODY FOLLICLE CELL DOMAINS
A signal from the oocyte specifies posterior FC iden-
tity, thus polarizing the anterior-posterior (A/P) axis of
the epithelium (González-Reyes et al., 1995; Roth et al.,
1995). Gurken (Grk) a TGF-␣-like ligand, is produced
in the oocyte (Neuman-Silberberg and Schüpbach,
1993). Both the EGF-receptor (EGFR) (Price et al.,
1989; Clifford and Schüpbach, 1992) and the Ras signal
transduction pathway are required in the FCs (Brand
and Perrimon, 1994; Hsu and Perrimon, 1994; Lee et
al., 1996). In the absence of EGF signaling, posterior
FCs express anterior FC markers (González-Reyes et
al., 1995; Roth et al., 1995). Conversely, global eleva-
tion of Ras activity early in oogenesis can induce ante-
rior FCs to transiently express the later posterior gene
expression marker, pointed (ptd) (Lee and Montell,
1997). Furthermore, expression of constitutively active
EGFR can induce anterior ptd expression and prevent
anterior decapentaplegic (dpp) expression (Larkin et
al., 1999). Thus, EGFR signaling prevents posterior
FCs from adopting the anterior fate.
The FCs at the ends of the egg chamber, the terminal
FCs, are distinct from FCs on the lateral surface of the
egg chamber, the mainbody FCs (Fig. 4). Terminal FCs
are uniquely competent to express posterior markers in
response to Ras activation. Global elevation of EGFR or
Ras activity does not induce ptd expression in the
mainbody FCs (Lee and Montell, 1997; Larkin et al.,
1999). Further studies led González-Reyes and St.
Johnston (1998a) to propose that each pair of polar
cells specifies a flanking domain of terminal cells (Fig.
4). Establishment of the terminal cells occurs by the
end of stage 6, is Notch dependent, and is independent
of A/P axis polarization (González-Reyes and St.
Johnston, 1998a; Larkin et al., 1999). Although termi-
nal specification is thought to precede A/P polarization,
the gene expression markers currently available for the
terminal cells reflect A/P patterning. These markers
are first detected during stage 7, and include slow
border cells (slbo) and dpp enhancer traps in the ante-
86 DOBENS AND RAFTERY
rior and ptd enhancer traps in the posterior (Lee and
Montell, 1997; Larkin et al., 1999).
Anterior FC become subdivided into multiple fates,
including the border cells, the squamous FCs and the
centripetally migrating FCs (González-Reyes et al.,
1995; Roth et al., 1995; Lee et al., 1996). González-
Reyes and St. Johnston (1998a) suggest that posterior
terminal cells are subdivided into equivalent domains
(Fig. 4), based on their observation that posterior cells
lacking the EGFR show position-specific expression of
anterior markers. These authors propose that each ter-
minal domain is patterned so that a series of concentric
competence domains surrounds the polar cells.
Whether anterior or posterior, all terminal cells at a
specific distance from the polar cells have a similar
positional identity, and are competent to give rise to
the appropriate anterior cell type. EGFR activation
then diverts posterior terminal cells to a posterior fate
(Fig. 4). Consistent with this proposed positional spec-
ification, the number and terminal position of the an-
terior border cells and squamous FCs are similar to the
number and terminal position of the two posterior cell
types that deposit the aeropyle (Fig. 1D, F). The cen-
tripetally migrating FCs were proposed as an outer-
most terminal domain (González-Reyes and St.
Johnston, 1998a); however, their identity as a terminal
cell type remains in question. Mainbody FCs can ex-
press a centripetal migration marker in response to
ectopic Dpp (Dobens and Raftery, 1998; Dobens et al.,
2000), suggesting that this cell type is an indirect re-
sult of terminal patterning.
EPITHELIAL REORGANIZATION DURING
MID-OOGENESIS
During stage 9, FCs reorganize into three physical
domains: a small cluster of border cells inside the egg
chamber, a squamous epithelium over the nurse cells,
and a columnar epithelium over the oocyte (Fig. 2). The
border cells detach from the anterior epithelium and
migrate through the nurse cells to reach the oocyte. At
the same time, mainbody FCs move posteriorly to form
a columnar epithelium over the oocyte. The remaining
30 – 40 anterior terminal cells flatten over the nurse
cells. Border cell migration and columnar reorganiza-
tion proceed in concert. Each migration involves many
of the same cell biological processes, so that some mu-
tations cause defects in multiple migrations (first noted
by Schüpbach and Wieschaus, 1986).
Border Cell Recruitment and Migration
During border cell migration, 5–10 FCs delaminate
from the anterior pole to invade the nurse cell complex
(Fig. 2C–F). The anterior polar cells recruit flanking
FCs to become border cells. Ectopic polar cells can
recruit additional border cells, but only in the anterior
half of the egg chamber (Liu and Montell, 1999). Con-
versely, failure to specify the posterior FC fate leads to
posterior expression of early border cell markers, such
as slbo (González-Reyes et al., 1995). Notch is impor-
tant for specification of border cells; in Nts egg cham-
bers, the presumptive border cells do not express slbo
and do not delaminate from the epithelium (González-
Reyes and St. Johnston, 1998a). Specification of the
border cell fate requires slbo, which encodes a bZIP
transcription factor (Montell et al., 1992). slbo is re-
quired for expression of border cell-specific genes in-
cluding the breathless (btl) FGF receptor and DE-cad-
herin (Niewiadomska et al., 1999).
Gratuitous Btl/FGF receptor expression rescues bor-
der cell migration in slbo mutants (Murphy et al.,
1995). Because the FGF receptor signals through the
Ras pathway, further studies used overexpression of
constitutively active or dominant negative Ras trans-
genes (Lee et al., 1996). These studies indicate that the
level and timing of Ras activity is critical to maintain
migration. Surprisingly, Slbo and Ras play antagonis-
tic roles during the initiation of migration (Lee et al.,
1996).
Migrating border cells form a highly organized clus-
ter, with the polar cells surrounded by a rosette of
flanking FCs. Specialized cell junctions appear to hold
the cluster together as it migrates between the nurse
cells (Niewiadomska et al., 1999). Migration involves
reorganization of the cytoskeleton; expression of dom-
inant negative Rac can block migration (Murphy and
Montell, 1996) and genes for cytoskeletal components
such as sqh and karst, are required (Edwards and
Kiehart, 1996; Zarnescu and Thomas, 1999). Notch
signaling may be active in migrating border cells, for
Notch protein levels are elevated (Xu et al., 1992).
Border cell delamination is regulated separately
from migration. discs large (dlg) inhibits delamination,
but not migration (Goode and Perrimon, 1997). Many
FCs migrate along the normal border cell route in dlg
mutants, even though they do not express border cell
markers. In contrast, ectopic border cells do not mi-
grate (Liu and Montell, 1999). They delaminate, only to
remain between the FC epithelium and the nurse cells.
These observations suggest that germ cells provide
guidance molecules for border cell migration.
DE-cadherin is essential for migration (Oda et al.,
1997; Niewiadomska et al., 1999); furthermore, the
speed of migration appears to be dependent on DE-
cadherin levels. Absence of DE-cadherin only in the
germ cells results in aberrant migration between the
FC epithelium and the nurse cells. The change in route
suggests that border cells preferentially adhere to the
cells with the highest level of DE-cadherin, indepen-
dent of tissue origin. Migration also requires DE-cad-
herin expression in the border cells. However, a cluster
only needs a few DE-cadherin-expressing cells to per-
mit migration. Strikingly, mosaic clusters that migrate
always have DE-cadherin positive cells at the leading
edge of migration. Taken together, these data indicate
that homophilic adhesion through DE-cadherin is in-
volved in both guidance and movement.
Mainbody Follicle Cell Reorganization
Posterior migration of the mainbody FCs is tightly
linked to formation of a columnar epithelium over the
 REGULATION OF FOLLICLE CELL MORPHOGENESIS
oocyte. Prior to stage 9, most FCs are cuboidal (Fig.
2A). Early in stage 9, FCs over the oocyte are columnar;
mainbody FCs become columnar as they move posteri-
orly to contact the oocyte (Fig. 2B). At the end of this
reorganization, a sharp apposition of columnar and
squamous cells lies directly over the oocyte-nurse cell
junction (Fig. 2C). Local cell-cell adhesion and signal-
ing between the germline and the FCs is critical to
reorganize the epithelium overlying the oocyte; both
DE-cadherin and Notch pathway genes are important
for this process. Surprisingly, the posterior migration
does not appear to involve actin-myosin-based motility
(Edwards and Kiehart, 1996), suggesting that this
movement is driven by other forces.
Formation of a columnar epithelium over the oocyte
may be regulated differently in the mainbody cells and
the posterior terminal cells. The Rho family of small
GTPase proteins regulates cell morphology in many
systems (Hall, 1998); their roles in FC morphogenesis
have been examined through the overexpression of
dominant negative and constitutively active trans-
genes (Murphy and Montell, 1996). Overexpression of
dominant negative RhoL results in loss of epithelial
structure specifically in the posterior terminal cells;
these FCs change shape and become multilayered. The
balance of Rho family GTPase activity may be critical,
for overexpression of constitutively active RhoL pro-
duces a similar phenotype. However, these observa-
tions do not clarify the function of Rho GTPases during
normal reorganization. Loss of ␣-spectrin also causes
loss of epithelial polarity specifically in posterior FCs
(Lee et al., 1997). These authors speculated that the
curvature of the egg chamber in the posterior is a
source of physical strain on the epithelium. In contrast,
the change in mainbody cell shape specifically requires
␤-H spectrin (Zarnescu and Thomas, 1999).
Notch is essential for posterior migration. Notch is
apically localized in the FCs, with higher levels in the
FCs over the oocyte than in the nurse cell-associated
FCs (Xu et al., 1992). A prolonged period with reduced
Notch function, using Nts, leads to a complete block of
reorganization; shorter periods result in migration of
fewer FCs, but otherwise normal reorganization of the
columnar epithelium (Xu et al., 1992). Reduced Notch
function can also cause specific loss of posterior epithe-
lial organization (Ruohola et al., 1991; Goode et al.,
1996; González-Reyes and St. Johnston, 1998a).
Cell-cell contacts with the germ cells are critical to
posterior migration. Reduction of DE-cadherin in the
germline causes a delay in posterior migration (Oda et
al., 1997). Similarly, germline expression of toc is es-
sential for posterior movement (Grammont et al.,
1997). In contrast, reduction of either egh or brn in the
germline causes a significant acceleration of posterior
migration, as well as aberrant FC shape (Goode et al.,
1996). When the oocyte is small, due to growth retar-
dation caused by fs(2)cup or Bicaudal D mutations, the
sharp demarcation between the squamous and colum-
nar epithelia is lost (Swan and Suter, 1996; Keyes and
Spradling, 1997). When the oocyte resides in a central
87
position, the overlying mainbody FCs become columnar
at stage 10 (González-Reyes and St. Johnston, 1998a).
All data support the model that contact with the oocyte
triggers elongation of posterior and mainbody FCs. The
resultant compaction of the posterior epithelium may
provide the force for posterior movement of the main-
body cells.
Formation of the Anterior
Squamous Epithelium
The flattening of anterior terminal FCs is a conse-
quence of patterning. A few ectopic squamous cells
form near the posterior pole when the posterior FC fate
is not specified (González-Reyes et al., 1995; Roth et al.,
1995). This indicates that the anterior cells are not
simply stretched as a secondary consequence of main-
body cell migration. It also indicates that a signal from
the nurse cells is not necessary for formation of the
overlying squamous epithelium. Little is known about
the mechanism of cell flattening. These cells remain
tightly associated with the nurse cells, thus they are
also called the nurse cell follicle cells. The nurse cell
FCs do not secrete chorion, and contract as the nurse
cells degenerate.
The current model for A/P patterning predicts that
absence of a posterior signal leads to flattening of the
anterior terminal cells. However, early over-expression
of activated Ras is not sufficient to prevent stage 10
anterior cell flattening. Instead, anterior cells can be-
come columnar at stage 10 when activated Ras is ex-
pressed in a slbo heterozygous background (Lee and
Montell, 1997). Similarly, small clones of posterior cells
that lack EGFR can express anterior markers, but the
cells retain columnar morphology (González-Reyes and
St. Johnston, 1998a). These data suggest that an ante-
rior signal is necessary for flattening. The BMP ligand
Dpp is a candidate for this signal, for it directs other
aspects of anterior FC patterning (Twombly et al.,
1996; Deng and Bownes, 1997; Dobens and Raftery,
1998; Dobens et al., 2000; Peri and Roth, 2000). Dpp is
expressed in the anterior terminal cells before and
during flattening (Fig. 5).
MORPHOGENESIS OF THE
EGGSHELL ARCHITECTURE
In the last stages of oogenesis, a tightly timed se-
quence of patterning interactions and migrations cre-
ates the elaborate architecture of the eggshell. We will
consider these final events in roughly sequential order.
Dorsal-Anterior Patterning of Columnar
Follicle Cells
Polarization of the dorsal-ventral (D/V) axis is in-
duced by a localized Grk signal from the oocyte (Schüp-
bach, 1987; Neuman-Silberberg and Schüpbach, 1993;
Roth et al., 1999). Prior to stage 7, the oocyte nucleus
resides in the extreme posterior of the cell. During
stages 7 and 8, the oocyte nucleus migrates to an asym-
metric position in the anterior. This repositioning is
essential for D/V patterning of the FCs and the result-
88 DOBENS AND RAFTERY

Fig. 4. Axial patterning of the follicle cells. Mainbody FCs are white,
terminal FCs are colored. Polar cells are determined in the germarium
(Margolis and Spradling, 1995). Establishment of terminal cells is Notch-
dependent, and occurs before stage 7 of oogenesis (González-Reyes
and St. Johnston, 1998a; Larkin et al., 1999). Subdivision of the terminal
cells into competence domains is proposed to occur before the anterior
and posterior fates are established (González-Reyes and St. Johnston,
1998a). The posterior follicle cell fate is induced by a Gurken signal from
the oocyte (González-Reyes et al., 1995; Roth et al., 1995). Patterning of
anterior-dorsal eggshell structures involves combinatorial signaling by
EGF and BMP signals (Dobens et al., 2000; Peri and Roth, 2000). The
eggshell diagram is color-coded to indicate the follicle cell population that
makes each structure.

Fig. 5. Flattening of the nurse cell follicle cells. Spectrin staining outlines the FCs (red), propidium iodide
stains nuclear DNA in blue; dpp enhancer trap (nuclear ␤-galactosidase; green) stains nuclei of prospective
squamous follicle cells in cuboidal epithelium at stage 8 (A). Dpp expression persists in these cells as they
flatten over the nurse cell complex during stage 9 (B) and persists through stage 10B (C).

ant embryo (recently reviewed in Ray and Schüpbach, The dorsal Grk signal is required to induce two
1996). The dorsal mainbody FCs are exposed to Grk prominent dorsal-anterior fates, the operculum and the
signal as they pass near the oocyte nucleus in the dorsal appendages. These eggshell structures are also
posterior migration. dependent on anterior patterning. When the oocyte is
 REGULATION OF FOLLICLE CELL MORPHOGENESIS
Fig. 6. Gene expression domains in the centripetally migrating cells.
In all panels, anterior is to the left; immunofluorescence staining reveals
nuclear ␤-galactosidase expression from enhancer traps in red, MYC
antigen expression used to mark FCs in green. A: Enhancer trap in dpp
marks nurse cell FCs at stage 10 in this confocal cross section (top);
dpp-expressing cells are at the leading edge of centripetal migration and
Fig. 7. Follicle cell migrations during dorsal appendage formation.
Panels A–C show immunofluorescence staining of egg chambers with
anti-␣-spectrin (red) to show cell outlines, and nuclear ␤-galactosidase
expressed from bunched-lacZ (green) marks elongating dorsal append-
age FCs prominently (arrows). DNA is stained by propidium iodide (blue).
89
come to lie adjacent to the forming micropyle at stage 12 (bottom). B:
A359 enhancer trap marks columnar FCs adjacent to Dpp-expressing
cells (top), which migrate to cover the anterior of the egg (bottom). C:
bunched enhancer trap expression is strictly excluded from the A359-
expressing cells (top) and abuts the edge of the operculum after centrip-
etal migration (bottom). Figures reprinted from Dobens et al., 2000.
small, the anterior mainbody FCs do not cover the
oocyte and dorsal appendages are aberrant or lacking
(Swan and Suter, 1996). Several EGFR target genes
are only expressed in response to the dorsal Grk signal
and not in response to the posterior signal (Brand and
Perrimon, 1994; Queenan et al., 1997; Sapir et al.,
1998). Hyperactivation of the EGFR/Ras signal trans-
duction pathway induces ectopic dorsal appendages
only in the anterior eggshell and ectopic gene expres-
sion only in the anterior mainbody FCs (Brand and
Perrimon, 1994). Two models are consistent with these
data. First, a terminal pre-pattern may specify the ring
of cells competent to form dorsal appendages. Second,
an anterior signal from the terminal cells may syner-
gize with the Grk signal to induce dorsal anterior fates.
Experiments in which the EGF-R/Ras signal trans-
duction pathway is activated in grk mutants support
the latter hypothesis. Dorsal target genes are ex-
pressed in a punctate pattern throughout the main-
body cells, and patches of dorsal appendage material
occur randomly throughout the eggshell (Queenan et
al., 1997; Sapir et al., 1998). Similarly, subtle eleva-
tions of EGFR signaling, for example in FC clones
lacking the EGF signaling antagonist Sprouty (Reich et
al., 1999), result in ectopic dorsal appendages only
along the collar (see also fs(1)K10, Heinlin et al., 1987).
90 DOBENS AND RAFTERY
The BMP ligand Dpp is a good candidate for a col-
laborating anterior signal. Dpp is expressed in the an-
terior terminal cells (Fig. 5; Twombly et al., 1996; Lar-
kin et al., 1999). High levels of Dpp expand the
operculum at the expense of the dorsal appendages
(Dobens et al., 2000). Moderate elevation of Dpp levels
can cause duplication of the dorsal appendages along
the edge of the collar (Twombly et al., 1996). The Dpp
type I receptor Thickveins and the BMP signal trans-
ducer Mad (Raftery and Sutherland, 1999) are required
to repress more posterior gene expression markers in
the anterior-most columnar follicle cells (Deng and
Bownes, 1997; Dobens et al., 2000). Intriguingly, Mad
is also required for expression of the Broad-Complex
genes in the dorsal appendage primordia (Peri and
Roth, 2000). Thus, it appears that combinatorial sig-
naling by BMP and EGF ligands generates the pattern
of anterior eggshell structures.
Competence for Centripetal Migration
At stage 10B, follicle cells migrate centripetally be-
tween the nurse cells and the oocyte, enclosing the
anterior of the egg (Fig. 2E, F). The centripetally mi-
grating FCs arise at the border between the anterior
squamous FCs and the columnar mainbody FCs, which
overlies the border between the nurse cells and the
oocyte (Fig. 2D). This migration occurs concurrently
with nurse cell dumping, so that the oocyte is capped by
FCs after the nurse cells have expelled their contents.
It is likely that the timing of centripetal migration is
critical for proper egg formation. If migration is com-
pleted too soon, nurse cell contents may not be com-
pletely transferred to the oocyte.
Competence to migrate centripetally is a conse-
quence of FC patterning. When the oocyte is small, the
junction between anterior terminal and mainbody FCs
lies over the nurse cell complex, rather than the nurse
cell-oocyte boundary. This sometimes leads to inappro-
priate migration between nurse cells, so that the egg-
shell encloses some nurse cells with the oocyte (Swan
and Suter, 1996). Distinct domains of gene expression
markers are visible in the anterior columnar FCs prior
to and during migration (Fig. 6) (Deng and Bownes,
1997; Dobens et al., 2000). In particular, Dpp is ex-
pressed in the cells that lead migration (Fig. 6A; Twom-
bly et al., 1996); the following cells express the A359
enhancer trap (Fig. 6B; Dobens et al., 2000). A359 is
expressed in a tight band of anterior columnar FCs, at
the boundary with the squamous FCs, prior to migra-
tion (Dobens and Raftery, 1998), thus marking the
presumptive centripetally migrating cells.
The bunched gene, which encodes a leucine zipper
transcription factor, acts in the mainbody FCs to limit
the size of this anterior domain (Dobens et al., 2000).
EGF signals stimulate bunched expression, but Dpp
represses bunched expression in anterior columnar
FCs (Fig. 6C). bunched inhibits the operculum fate in
more posterior mainbody FCs, and regulates expres-
sion of cytoskeletal components involved in migration
(LLD and LAR, unpublished data). Thus, a posteriorly-
expressed leucine zipper transcription factor, Bunched,
antagonizes anterior patterning by Dpp (Dobens et al.,
2000). Similarly, an anteriorly-expressed leucine zip-
per transcription factor, Slbo, antagonizes posterior
patterning directed by activated Ras (Lee et al., 1996).
Centripetal migration involves DE-cadherin medi-
ated recognition between the FCs and the oocyte. Lack
of DE-cadherin in the germline results in FC migration
between the nurse cells (Oda et al., 1997; Niewiadom-
ska et al., 1999). Migration also is disrupted when FCs
lacking DE-cadherin are in or near the presumptive
centripetally migrating FCs (Niewiadomska et al.,
1999). Migration requires myosin II, which accumu-
lates at the apical tips of centripetally migrating cells
to form a supracellular ring (Edwards and Kiehart,
1996). Notch accumulates in high levels in the migrat-
ing cells, and centripetal migration can be blocked in
Nts experiments (Xu et al., 1992).
The cells at the leading edge of centripetal migration
come to rest at the border cells. The leading edge cells
express Dpp during the migration and throughout mi-
cropyle formation (Fig. 6A; Twombly et al., 1996; Do-
bens et al., 2000). Although centripetal migration is
radially symmetrical, the micropyle is situated at the
ventral edge of the mature operculum (Fig. 3E). For-
mation of the operculum is directed by both EGF and
Dpp signals. High levels of Dpp expand an otherwise
normal operculum (Twombly et al., 1996). Overexpres-
sion of an activated EGFR can expand the operculum
material, but it is aberrantly patterned and lacks a
collar (Queenan et al., 1997). bunched integrates these
signals to pattern the collar structure (Dobens et al.,
2000). Together, the leading edge cells and the border
cells create the micropyle of the eggshell. The leading
edge cells deposit the vitelline membrane and endocho-
rion, whereas the border cells extend a process that
forms the canal (Edwards et al., 1997).
Formation of the Dorsal Appendages
Continued EGF ligand signaling is required for for-
mation of the dorsal appendages. However, exposure to
the Grk signal is cut off after vitelline membrane dep-
osition, which begins at stage 10. The dorsal Grk signal
induces local FC expression or activation of two addi-
tional EGFR ligands, Vein and Spitz (Sapir et al., 1998;
Wasserman and Freeman, 1998). The two dorsal ap-
pendage primordia separate when peak EGFR signal-
ing at the dorsal anterior midline induces expression of
an EGFR inhibitor, Argos (Wasserman and Freeman,
1998). These EGFR feedback loops have been reviewed
in detail (Stevens, 1998). Argos modulation of EGFR
signaling creates the patch of type III operculum cells
that separates the dorsal appendages (Fig. 3E). High
level EGFR signaling continues in two dorso-lateral
patches that will migrate to form the dorsal append-
ages during stages 11 through 14.
The dorso-lateral patches of FCs migrate anteriorly
over the nurse cell FCs (Fig. 7) and form two stretching
tubes. This process is easily perturbed, causing dorsal
appendage fusions and shortening. FC patterning is
 REGULATION OF FOLLICLE CELL MORPHOGENESIS
critical; germline expression of bullwinkle is also re-
quired (Rittenhouse and Berg, 1995). Dorsal append-
age formation may be indirectly perturbed when nurse
cell dumping is blocked (Robinson and Cooley, 1997).
However, both nurse cell dumping and dorsal append-
age formation involve cytoskeletal rearrangements, so
that some genes may be independently required in both
processes. For example, the requirement for myosin
light chain in dorsal appendage formation can be sep-
arated in time from the requirement in nurse cell
dumping (Edwards and Kiehart, 1996). JUN kinase
signaling plays a role in dorsal appendage morphogen-
esis (S. Noselli, personal communication). In contrast,
the JUN kinase phosphatase gene puckered acts in the
centripetally migrating cells (LLD, A. Martin-Blanco,
A. Martinez-Arias, F. Kafatos, and LAR, unpublished
data). These observations suggest that the level of Jun
kinase signal transduction is important for orchestrat-
ing each of these migrations.
PERSPECTIVE
Dissecting the molecular pathways that specify cell
fate and coordinate movement requires a well-defined
and simple tissue. The follicle cells provide a geneti-
cally tractable system, where mosaic analysis can es-
tablish the molecules that are necessary for an event
and clonal overexpression can establish the molecules
that are sufficient to induce it. Egg chambers will de-
velop in organ culture, so that genetic analysis can be
coupled with in vitro approaches. Through these tech-
nical advantages, Drosophila follicle cells offer a
unique opportunity to understand the cell biology of
epithelial morphogenesis.
The architecture of the follicle cell epithelium is
achieved through successive iterations of local induc-
tive interactions between tissue layers, regional pat-
terning signals from within the epithelium, and con-
certed migrations of specific cell types. The regulatory
pathways for these events are the same pathways that
pattern other organs in both invertebrates and verte-
brates. Notch signaling and cadherin-mediated adhe-
sion are involved in many of the morphogenetic events.
Late patterning of the FCs involves combinatorial BMP
and EGFR signaling.
Key questions that remain for FC morphogenesis
reflect important questions for the field of developmen-
tal biology. Morphogenesis of the FCs appears to in-
volve temporal changes in competence in addition to
positional signaling. Are these temporal changes due to
a required sequence of patterning events or does an
internal clock regulate events in the FCs? The timing of
nurse cell dumping is regulated by the level of ecdysone
produced by the germ cells (Buszczak et al., 1999).
Ecdysone signaling may regulate dorsal appendage for-
mation as well (Tzolovsky et al., 1999). It will be im-
portant to determine whether this signal acts as an
internal clock to coordinate morphogenesis of the two
cell types.
The evidence for subdivision of epithelial FCs into
terminal and mainbody cell types is compelling. How-
91
ever, the sequence and timing of subsequent pattern-
ing events remains unclear. Are terminal regions cryp-
tically subdivided by an early positional signal, or do
later interactions at the terminal-mainbody cell bound-
aries organize new fates? Boundary formation is im-
portant in creating organizing centers for morphogen-
esis of other tissues (Dahmann and Basler, 1999); do
cell type boundaries serve as organizing centers in the
FC? Cell sorting has been proposed to be important at
boundaries (Dahmann and Basler, 1999), and many FC
migrations involve selective adhesion between cells ex-
pressing high levels of DE-cadherin. Do boundaries
between follicle cell types play a role in initiating mi-
grations? In other tissues, Notch signaling plays a
prominent role in creating organized boundaries be-
tween two cell types (Irvine, 1999). Understanding the
multiple roles of Notch in FC patterning will be impor-
tant to resolve this question.
ACKNOWLEDGMENTS
We thank W. Petri and L. Margaritis for generously
sharing electron micrographs of eggshell structures, S.
Noselli for discussing unpublished results, and D.
Branton for antisera. We are grateful to Flybase and
The Interactive Fly for providing organized and up-to-
date information on the Web. Members of the Raftery
and Perrimon labs provided useful discussions. The
Raftery lab is supported by grants from the American
Cancer Society (RPG-96-103-03-DDC) and NIH
(GM60501).
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