spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

doi: 10.1242/10.1242/dev.00533

This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Related articles in Development
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Urbach, R.
Right arrow Articles by Technau, G. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Urbach, R.
Right arrow Articles by Technau, G. M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Molecular markers for identified neuroblasts in the developing brain of Drosophila

Rolf Urbach and Gerhard M. Technau

Institut für Genetik, Universität Mainz, D-55099 Mainz, Germany



View larger version (102K):

[in a new window]
  		Fig. 1. Expression of gap genes at embryonic stages 9 (st9) and 10 (st10). Semi-schematic presentations of the left half of head flat preparations [anterior (a) is towards the top; dorsal (d) is towards the left]. Colour code indicates protein expression patterns of Empty spiracles (ems), Hunchback (hb), Orthodenticle (otd), Sloppy paired 1 (slp1), and the huckebein-lacZ (hkb) and tailless-lacZ (tll) patterns in identified brain neuroblasts [A,E; colour intensity reflects weak (~) and strong (+) expression] and the peripheral procephalic ectoderm (B-D; F-H; thick hatching indicates strong expression, thin hatching indicates weak expression) at stage 9 (A-D) and stage 10 (E H). Nomenclature of brain NBs is according to their position in the trito- (T), deuto- (D) and protocerebrum [P; for details of the nomenclature see Urbach et al. (Urbach et al., 2003Go)]. Red lines in A,E mark the boundaries between trito-, deuto- and protocerebrum. In B,F, ems ectodermal domains arising from the blastodermal ems stripe are numbered (1-4; compare with Fig. 2N). In D,H, high level and low level tll-lacZ expression domains correspond to HL and LL in Rudolph et al. (Rudolph et al., 1997Go). Note, that slp1 is segmentally expressed in the intercalary (slp1 is, slp1 intercalary stripe) and antennal (slp1 as, slp1 antennal stripe) ectoderm. Stars indicate mandibular NBs. Engrailed ectodermal domains (encircled in red) are shown for comparison. as, en antennal stripe; CL, clypeolabrum; FG, forgut; hs, en head spot; is, en intercalary stripe; ML, ventral midline; slp1 oc, ocular slp1 domain; slp1 lr, labral slp1 domain.

 


View larger version (119K):

[in a new window]
  		Fig. 2. Expression of gap genes at embryonic stage 11 (st11). (A-L) Left half of head flat preparations double stained for expression of a gap gene and en/inv (lacZ or antibody staining) as indicated. Pictures on the left (A,C,E,G,I,K) focus on the peripheral procephalic ectoderm; broken lines indicate the ventral midline; insets depict lateral views of stage 11 whole-mount heads. Middle panels (B,D,F,H,J,L) show close-up views of regions indicated in A,C,E,G,I,K by black frames; the focus is at the level of NBs. Positively stained NBs are indicated by white labels, negative ones are indicated by black labels. (A,B) Ems/en-lacZ. (C,D) Hb/en-lacZ. (E,F) hkb-lacZ/Inv. (G,H) Otd/en-lacZ. Note that Otd is expressed in the ventral midline, in a small domain within the ventral intercalary ectoderm (red arrow) and in cells along the dorsal midline (red arrowheads). (I,J) tll-lacZ/Inv; high level (HL) and low level (LL) tlllacZ-expression is indicated. (K,L) Slp1/en-lacZ. (M) Summary of gap gene expression in brain NBs as indicated by the colour code, which distinguishes between weak (~) and strong (+) expression of tll- and hkb-lacZ, and Slp1. Red lines mark the boundaries between tritocerebrum, deutocerebrum, ocular and putative labral neuromer. Stars mark mandibular NBs. (N-P) Semi-schematic presentation of gap gene expression in the peripheral procephalic ectoderm. (N) ems ectodermal domains are numbered (1-5), 3a and 3b derive from intercalary ems domain 3 (compare with Fig. 1B,F). Anterior (a) is towards the top; dorsal (d) is towards the left. AN, IC, MD, MX, antennal, intercalary, mandibular and maxillary segment, respectively; CL, clypeolabrum; FG, foregut; ML, ventral midline; OA, Bolwig organ/optic lobe anlagen; as, en antennal stripe; cl, en expression in the clypeolabrum; hs, en head spot; is, en intercalary stripe; mds, en mandibular stripe; slp1 as, slp1 antennal stripe; slp1 is, slp1 intercalary stripe; slp1 oc, ocular slp1 domain; slp1 lr, labral slp1 domain.

 


View larger version (120K):

[in a new window]
  		Fig. 3. Expression of the pair rule gene runt and the homeotic gene labial at embryonic stages 9, 10 and 11. Semi-schematic presentation of the Runt and Labial protein (Lab) patterns in brain NBs (A,C,E) and outer procephalic ectoderm in relation to the en pattern (B,D,F) at stage 9 (st9, A,B), stage 10 (st10, C,F) and stage 11 (st11, E,F) as indicated; numbers (1-3) in B,D,F reflect the temporal sequence of the ectodermal runt domains. Lab expression covers the complete intercalary neuroectoderm and emerging NBs. (E) Note that all trito- and two deutocerebral NBs express Lab; red lines demarcate neuromeric boundaries. (G,H) en-lacZ/Lab double stained head flat preparation. (G) Lab expression covers the en intercalary stripe (is) and partially the en antennal stripe (as; compare with F); broken line marks the anterior boundary of the Lab domain. (H) Higher magnification of the region indicated by black frame (in G) focusing at the level of NBs. Note that the Lab-positive Dv4 is at the same anteroposterior level as the En-positive Dv8. (I,J) en-lacZ/Runt double labelled head flat preparation. (I) In the periphery, Runt expression is confined to the ocular (1) and intercalary (3) ectodermal domain. (J) Higher magnification of region marked by black frame in I, focusing at the level of NBs. Positively stained NBs in H,J are indicated by white labels, negative ones are indicated by black labels; insets in G,I show a lateral view of stage 11 wholemounts; dashed line marks the ventral midline. Anterior (a) is towards the top; dorsal (d) is towards the left. AN, IC, MD, MX, antennal, intercalary, mandibular and maxillary segment, respectively; CL, clypeolabrum; FG, foregut; ML, ventral midline; as, en antennal stripe; hs, en head spot; is, en intercalary stripe.

 


View larger version (137K):

[in a new window]
  		Fig. 4. Expression of the early eye genes dac, ey and toy at embryonic stages 9, 10 and 11. Semi-schematic presentation of Dachshund (dac), Eyeless (ey), and Twin of eyeless (toy) expression in brain NBs [A-C; low (~) and high (+) expression levels; red lines indicate neuromeric boundaries], and within the outer procephalic ectoderm (D-F; thick hatching indicates strong expression, thin hatching indicates weak expression) as indicated at stage 9 (st9, A,D), stage 10 (st10, B,E), and stage 11 (st11, C,F). Engrailed (en) expression domains in the peripheral ectoderm are indicated for comparison. (G-L) Double staining for expression of svp-lacZ (G-J) or en-lacZ (K,L), and Dachshund (dac; G,H), Eyeless (ey; I,J) and Twin of eyeless (toy; K,L). (G,I,K) Outer procephalic ectoderm of flat preparations (dashed line indicates ventral midline); insets show lateral views of stage 11 wholemounts. (H,J,L) Higher magnifications of regions indicated by frames in G,I,K at the level of brain NBs. (G) Red arrow indicates the intercalary Dac-positive ectodermal domain including the Dac-positive NB Tv2. (I,J) Ey is expressed in few NBs; Ey/en-lacZ double labelling (see inset on bottom left in I) shows Ey expression in the intercalary and antennal head immediately anterior to the respective en domains (arrows). (K,L) Toy is mainly confined to protocerebral NBs and to Dd8. Anterior (a) is towards the top; dorsal (d) is towards the left (except for insets). AN, IC, MD, MX, antennal, intercalary, mandibular and maxillary segment, respectively; CL, clypeolabrum; FG, foregut; ML, ventral midline; OA, Bolwig organ/optic lobe anlagen; as, en antennal stripe; hs, en head spot; is, en intercalary stripe.

 


View larger version (95K):

[in a new window]
  		Fig. 7. Summary diagram of molecular markers expressed in brain NBs at different stages of embryonic development. Colour code indicates the expression of more than 40 different molecular markers (representing 34 different genes as listed) in individual brain NBs. In addition to the markers described in this paper, these maps include the expression of the proneural, segment polarity, DV patterning and glial marker genes, a detailed description of which is given in the accompanying papers (Urbach et al., 2003Go; Urbach and Technau, 2003Go). Each brain NB expresses a specific combination of molecular markers. Accordingly, each NB can be uniquely identified at stage 9 (A; st9), stage 10 (B; st10) and stage 11 (C; st11, which is when the entire population of brain NBs is established). Red lines indicate the neuromeric boundaries between the trito- (T) and deutocerebrum (D), and the ocular (Oc-P) and labral protocerebrum (Lr-P). Anterior (a) is towards the top; dorsal (d) is towards the left. CL, clypeolabrum; FG, foregut.

 


View larger version (84K):

[in a new window]
  		Fig. 5. Expression of cas, Fas2, klu, lbe, pdm1 and unpg at embryonic stages 9, 10 and 11. Semi-schematic presentation of Castor (cas), Fasciclin 2 (Fas2), Klumpfuss (klu), Ladybird early (lbe), POU-domain 1 (pdm1) protein, and unplugged-lacZ (unpg) expression in brain NBs [A,D,G; low (~) and high (+) expression levels; red lines mark neuromeric boundaries], and within the outer procephalic ectoderm [B,C,E,F,H,I; thick hatching indicates strong (+), thin hatching weak (~) expression] as indicated at stage 9 (st9, A-C), stage 10 (st10, D-F) and stage 11 (st11, G-I). Engrailed (en) expression domains in the peripheral ectoderm are indicated for comparison. Anterior (a) is towards the top; dorsal (d) is towards the left. AN, MD, MX, antennal, mandibular and maxillary segment, respectively; CL, clypeolabrum; FG, foregut; ML, ventral midline; OA, Bolwig organ/optic lobe anlagen; as, en antennal stripe; hs, en head spot; is, en intercalary stripe.

 


View larger version (172K):

[in a new window]
  		Fig. 6. Expression of cas, Fas2, klu, lbe, pdm1 and unpg at embryonic stage 11. Double staining for expression of En/Inv (lacZ or antibody staining) or svp-lacZ (C,D) and Castor (cas; A,B), Fasciclin 2 (Fas2; C,D), Klumpfuss (klu; E,F), Ladybird early (lbe; G,H), Pou-domain 1 (pdm1; I,J), and unplugged-lacZ (unpg-lacZ; K,L), respectively, as indicated. (A,C,E,G,I,K) Focus is on the outer procephalic ectoderm of flat preparations (dashed line indicates ventral midline); insets show lateral views of wholemounts. (B,D,F,H,J,L) Higher magnifications of regions indicated by frames in A,C,E,G,I,K at the level of deuto- and protocerebral NBs; positively stained NBs are indicated by white labels, negative ones by black labels; NBs in brackets are not in focus. (A) Red arrowhead indicates expression of Cas in the outer ectoderm; dashed red line indicates the boundary between the labral (Lr-P) and ocular part of the protocerebrum (Oc-P). Cas is prominently expressed in most brain NBs (B). (C,D) Fas2 is expressed on the surface of most tritocerebral NBs and a few deutocerebral NBs (D). (E,F) Nuclear Klu expression is detected in almost all brain NBs. (G,H) Lbe is segmentally expressed in the head (arrows). (I,J) Approximately one half of the brain NBs express pdm-1, including most deuto- and tritocerebral, as well as central ocular and part of the labral, NBs. (K,L) unpg-lacZ is strongly expressed in almost all deutocerebral NBs, and in some ocular NBs close to the deutocerebral/ocular border. AN, antennal, segment; CL, clypeolabrum; Lr-Pc, labral part of the protocerebrum; Oc-Pc, ocular part of the protocerebrum; OA, Bolwig organ/optic lobe anlagen; as, en antennal stripe; hs, en head spot; is, en intercalary stripe. Anterior (a) is towards the top; dorsal (d) is towards the left.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2003