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Int. J. Environ. Res. Public Health 2017, 14(2), 213; doi:10.3390/ijerph14020213

Characterization of Carbonic Anhydrase 9 in the Alimentary Canal of Aedes aegypti and Its Relationship to Homologous Mosquito Carbonic Anhydrases

1
The Whitney Laboratory, University of Florida, Saint Augustine, FL 32080, USA
2
The Anastasia Mosquito Control District, St. Augustine Florida, Saint Augustine, FL 32092, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Paul B. Tchounwou
Received: 20 December 2016 / Revised: 13 February 2017 / Accepted: 14 February 2017 / Published: 21 February 2017
(This article belongs to the Special Issue Mosquito Control Innovations into The 21st Century)
View Full-Text   |   Download PDF [4827 KB, uploaded 21 February 2017]   |  

Abstract

In the mosquito midgut, luminal pH regulation and cellular ion transport processes are important for the digestion of food and maintenance of cellular homeostasis. pH regulation in the mosquito gut is affected by the vectorial movement of the principal ions including bicarbonate/carbonate and protons. As in all metazoans, mosquitoes employ the product of aerobic metabolism carbon dioxide in its bicarbonate/carbonate form as one of the major buffers of cellular and extracellular pH. The conversion of metabolic carbon dioxide to bicarbonate/carbonate is accomplished by a family of enzymes encoded by the carbonic anhydrase gene family. This study characterizes Aedes aegypti carbonic anhydrases using bioinformatic, molecular, and immunohistochemical methods. Our analyses show that there are fourteen Aedes aegypti carbonic anhydrase genes, two of which are expressed as splice variants. The carbonic anhydrases were classified as either integral membrane, peripheral membrane, mitochondrial, secreted, or soluble cytoplasmic proteins. Using polymerase chain reaction and Western blotting, one of the carbonic anhydrases, Aedes aegypti carbonic anhydrase 9, was analyzed and found in each life stage, male/female pupae, male/female adults, and in the female posterior midgut. Next, carbonic anhydrase 9 was analyzed in larvae and adults using confocal microscopy and was detected in the midgut regions. According to our analyses, carbonic anhydrase 9 is a soluble cytoplasmic enzyme found in the alimentary canal of larvae and adults and is expressed throughout the life cycle of the mosquito. Based on previous physiological analyses of adults and larvae, it appears AeCA9 is one of the major carbonic anhydrases involved in producing bicarbonate/carbonate which is involved in pH regulation and ion transport processes in the alimentary canal. Detailed understanding of the molecular bases of ion homeostasis in mosquitoes will provide targets for novel mosquito control strategies into the new millennium. View Full-Text
Keywords: carbonic anhydrase (CA); mosquito; alkaline; pH; midgut; immunohistochemistry; data mining; molecular phylogenetics carbonic anhydrase (CA); mosquito; alkaline; pH; midgut; immunohistochemistry; data mining; molecular phylogenetics
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Supplementary materials

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    Doi: 122016
    Link: http://www.mdpi.com/link,Figure S1:Alignment of AAEL005520 splice variants; Figure S2:Alignmnet of AAELO10893 splice variants
    Description: Supplementary Figure 1. Alignment of AeCA3 splice variants. This figure shows only the alignments of the 5’ and 3’ UTR. The nucleotide sequences highlighted in black are identical while nucleotide sequences highlighted in grey are similar. The 5’ UTR is indicated by a magenta box, while the 3’ UTR is indicated by a light blue boc. The 5’ UTRs between both variants are divergent except for the last 83 bp. Supplementary Figure 2. Alignment of AeCA7 splice variants. This figure shows only the alignments of the 5’ and 3’ UTR. The nucleotide sequences highlighted in black are identical while nucleotide sequences highlighted in grey are similar. The 5’ UTR is indicated by a magenta box while the 3’ UTR is indicated by a light blue box. The 5’ UTRs between both sequences are not similar to each other.

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MDPI and ACS Style

Dixon, D.P.; Van Ekeris, L.; Linser, P.J. Characterization of Carbonic Anhydrase 9 in the Alimentary Canal of Aedes aegypti and Its Relationship to Homologous Mosquito Carbonic Anhydrases. Int. J. Environ. Res. Public Health 2017, 14, 213.

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