A light-dependent ammonia-assimilating mechanism in the ctenidia of a giant clam

Hiong, Kum C.; Choo, Celine Yen Ling; Boo, Mel V.; Ching, Biyun; Wong, Wai P.; Chew, Shit F.; Ip, Yuen K.

doi:10.1007/s00338-016-1502-4

Cited by 40 publications

(46 citation statements)

References 33 publications

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“…PCR and cloning experiments were performed as described previously in Hiong et al. (). Analysis of multiple clones of CA2‐like fragments from the outer and inner mantle of T. squamosa revealed only one form.…”

Section: Methodsmentioning

confidence: 99%

“…The qPCR reactions contained 5 μ L of 2X Fast SYBR ® Green Master Mix (Applied Biosystems), 0.3 μ mol L −1 of forward and reverse primers each, and various quantities of standard (to construct the standard curve) or 1 ng of sample cDNA in a total volume of 10 μ L. The cycling conditions, melt curve analysis, and construction of a standard curve were performed according to the method of Hiong et al. (,b). The amplification efficiency of the CA2‐like primers was 97.4%.…”

Section: Methodsmentioning

confidence: 99%

“…Protein extraction and SDS‐PAGE were performed according to the methods of Hiong et al. (,b). Fifty micrograms of protein were electrophoretically separated and transferred onto PVDF membranes.…”

Section: Methodsmentioning

confidence: 99%

“…Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society gigas CA2 (XM_011449590.2), Mytilus galloprovincialis CA2 (KT818923.1), Octopus bimaculoides CA2-like (XM_014929272.1), and Danio rerio CA2 (NM_199215.1). PCR and cloning experiments were performed as described previously in Hiong et al (2017a). Analysis of multiple clones of CA2-like fragments from the outer and inner mantle of T. squamosa revealed only one form.…”

Section: Pcr Cloning and Race-pcrmentioning

confidence: 99%

“…) and glutamine synthetase (Hiong et al. ) in the ctenidium, would be upregulated by light exposure. In order to shed light on the possible physiological function of CA2‐like in T. squamosa , immunofluorescence microscopy was also performed to confirm its cellular localization.…”

Section: Introductionmentioning

confidence: 99%

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Carbonic anhydrase 2‐like in the giant clam, Tridacna squamosa : characterization, localization, response to light, and possible role in the transport of inorganic carbon from the host to its symbionts

Koh

Hiong

et al. 2017

Physiol Rep

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The fluted giant clam, Tridacna squamosa, lives in symbiosis with zooxanthellae which reside extracellularly inside a tubular system. Zooxanthellae fix inorganic carbon (Ci) during insolation and donate photosynthate to the host. Carbonic anhydrases catalyze the interconversion of CO 2 and HCO3−, of which carbonic anhydrase 2 (CA2) is the most ubiquitous and involved in many biological processes. This study aimed to clone a CA2 homolog (CA2‐like) from the fleshy and colorful outer mantle as well as the thin and whitish inner mantle of T. squamosa, to determine its cellular and subcellular localization, and to examine the effects of light exposure on its gene and protein expression levels. The cDNA coding sequence of CA2‐like from T. squamosa comprised 789 bp, encoding 263 amino acids with an estimated molecular mass of 29.6 kDa. A phenogramic analysis of the deduced CA2‐like sequence denoted an animal origin. CA2‐like was not detectable in the shell‐facing epithelium of the inner mantle adjacent to the extrapallial fluid. Hence, CA2‐like is unlikely to participate directly in light‐enhanced calcification. By contrast, the outer mantle, which contains the highest density of tertiary tubules and zooxanthellae, displayed high level of CA2‐like expression, and CA2‐like was localized to the tubule epithelial cells. More importantly, exposure to light induced significant increases in the protein abundance of CA2‐like in the outer mantle. Hence, CA2‐like could probably take part in the increased supply of inorganic carbon (Ci) from the host clam to the symbiotic zooxanthellae when the latter conduct photosynthesis to fix Ci during light exposure.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Pcr Cloning and Race-pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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