Article Figures & Tables
Figures
- Fig. 1.
(A) A living fourth instar Anopheles gambiae larva that had been fed pH sensitive dye (m-cresol purple) to highlight the gut luminal pH gradient. The structure and compartments of the alimentary canal are outlined in black. (B) A stylized diagram of the gut with the anterior to the left. The brackets above indicate the extent of each region, and the approximate pH of each is indicated (modified from Neira Oviedo et al., 2008). GC, gastric caeca; AMG, anterior midgut; PMG, posterior midgut; HG, hindgut; MT, Malpighian tubules; CM, caecal membrane; PT, peritrophic matrix.
- Fig. 2.
Confocal immunofluorescence microscopy to identify GPI-linked carbonic anhydrase 10 (CA10) in isolated whole-mount preparations of Anopheles gambiae fourth instar larval gut. (A) A low magnification view (maximum projection of a stack of images) of the anterior half of the midgut. CA10 is labeled in green and muscle actin (phalloidin) in red. This panel is an overlay of the two color channels and therefore red indicates peripheral muscles of the gut and yellow indicates simultaneous labeling for CA10 and muscle actin. Note lateral complexes of muscles that are double labeled and central muscles (in this view dorsal) that are labeled for actin only. (B) A single color channel (green; CA10) at high magnification. Arrow indicates specific muscles that label for CA10. (C) The same view but with only the red color channel (phalloidin; muscle) shown. (D) Cell nuclei labeled with DRAQ5 (blue; nuclei). (E) An overlay of all three color channels demonstrating that specific gut muscles are labeled for CA10 and others are not. (F) A single plane of a stack of images at higher magnification, showing the muscle cell surface location of the CA10 labeling (green). GC, gastric caeca; AMG, anterior midgut. Arrows in B,C and E indicate the same point in the CA10-positive muscles (modified from Seron et al., 2004). Scale bars, 80 μm (A); 20 μm (B–E); 30μ m (F).
- Fig. 3.
Molecular phylogenetic analysis of carbonic anhydrase (CA) gene sequences from Homo sapiens (black), Anopheles gambiae (green), Aedes aegypti (blue) and Drosophila melanogaster (red) (Smith et al., 2007).
- Fig. 4.
Confocal immunofluorescence microscopy to identify AgCA9 in sections and whole mounts of Anopheles gambiae larval gut tissues. (A) A longitudinal section of a fourth instar larva labeled for AgCA9 (green) and Na+/K+-ATPase (red). Note prominent AgCA9 labeling in the cells and lumen of the gastric caeca, the ectoperitrophic fluid and the anterior portion of the rectum. (B,C) Two views of a whole-mount preparation of larval rectum showing AgCA9 (green; B) and Na+/K+-ATPase (C). (D) A high magnification cross-section of the rectum with the anterior to the right showing the mutually exclusive labeling for CA-9 (green) and Na+/K+-ATPase (red). The dorsal anterior rectum (DAR) cells are indicated by the arrow. (E) A high magnification view of an isolated rectum with the DAR cells (arrow) labeled for AgCA9 and the external rectal musculature labeled in red (phalloidin). GC, gastric caeca; AMG, anterior midgut; PMG, posterior midgut; MT, Malpighian tubules. All arrows indicate the DAR cells. Scale bars, 150 μm (A–C), 75 μm (D,E).
- Fig. 5.
Analyses of the members of the SLC4A family of anion exchangers/transporters. (A) A molecular phylogenetic tree of the annotated members of this gene family from Anopheles gambiae (green), Aedes aegypti (blue), Drosophila melanogaster (red) and Homo sapiens (black). The human genes include putative and confirmed splice variants of several genes. Similarly, several putative splice variants of the Drosophila genes are included. The tree was generated using MrBayes, 1.5 million iterations and the JTT amino acid substitution model. All nodes represent at least 95% probability but the branch lengths are arbitrary. (B) Hydropathy plots generated for Drosophila NDAE1 (black) and Anopheles AgAE1 (blue). (C) The hypothetical secondary structure of one putative splice variant of AgAE1.
- Fig. 6.
A DNA microarray-based analysis of regionalized expression of the three annotated SLC4A gene family members in Anopheles gambiae larvae. Whole larval gut expression (W) is compared with transcriptome levels in the gastric caeca (GC), anterior midgut (AMG), the posterior midgut (PMG) and the Malpighian tubule/hindgut (HG) (Neira Oviedo et al., 2008). A color scale indicates the transcriptome levels.
- Fig. 7.
Partial model of CA distribution in gut (upper drawing) and physiological impact (lower drawing). Blue arrows indicate hypothetical diffusion of metabolic CO2, only indicated in AMG cells which lack anα -CA. V, V-ATPase; Na/K, Na+/K+-ATPase; NHA, Na+/H+ antiporter; CA, carbonic anhydrase.
Tables
- Table 1.
Carbonic anhydrase genes annotated in the Anopheles gambiae genome
ENSANGP000000 ENSANGT000000 ENSANGG000000 New convention GenBank accession no. α-CA 01278 01278 01096 AgCA1 CA-RP 01574 01574 01335 AgCA-RP2 α-CA 10017 10017 07528 AgCA3 α-CA 11013 11013 08524 AgCA4 CA-RP 11908 11908 09419 AgCA-RP5 α-CA 12957 12957 10468 AgCA6 DQ518577 α-CA 14919 14919 12430 AgCA7 α-CA 18999 18999 16510 AgCA8 α-CA 19212 19212 16723 AgCA9 DQ518576 α-CA 21313 21313 18824 AgCA10 AY280612 α-CA 21739 21739 19250 AgCA11 AY280613 β-CA 29115 28335 15071 AgCAb EF065522 -
Two of the α-CAs are CA-related proteins (CA-RPs). One β-CA is also listed. The Ensembl annotation numbers for the coded proteins (second column from left), transcript (third column from left) and gene (fourth column from left) are given along with the new notation for the genes as given by Smith et al. (Smith et al., 2007). The GenBank accession numbers are shown in the final column on the right for each transcript that has been fully cloned by the Linser laboratory
-
Article navigation
Related articles
Cited by...
More in this TOC section
Similar articles
Other journals from The Company of Biologists
Meet the team at SICB – 3-7 January 2020
Meet the Journal of Experimental Biology team at the SICB 2020 meeting in Austin, Texas! News & Views Editor Kathryn Knight and Reviews Editor Stefan Galander will be at booth 507, along with a selection of JEB goodies including the 2020 Calendar, 2019 Highlights booklet and new JEB T-shirts.
Springy bamboo poles help villagers carry more than their own body weight
People in Southeast Asia often carry extremely heavy loads suspended from a bouncy bamboo pole slung over one shoulder. It turns out that they adjust their stride to bounce in time with the pole, allowing them to save 20% of their energy. Read more.
Two-year pilot transitional open access agreement
We are excited to announce a two-year pilot transitional open access agreement with Jisc from January 2020. Researchers at participating institutions will have unlimited access to The Company of Biologists’ three subscription journals and will be able to publish their research papers open access. Find out more.
Commentary — The utility and determination of Pcrit in fishes
Gordon Ultsch and Matthew Regan outline why Pcrit is a useful and informative comparator of hypoxia tolerance in fishes, provided it is determined using standardized respirometry methods and sound statistical approaches.
Where will your research take you?
Early-career researchers can apply for up to £2,500 to offset the cost of travel and expenses to make collaborative visits to other labs around the world. Read about Pierre’s experience in Greenland, where he continued research into the Greenland Shark’s remarkable longevity.