PMID- 25766383 OWN - NLM STAT- MEDLINE DCOM- 20160108 LR - 20181202 IS - 1532-0456 (Print) IS - 1532-0456 (Linking) VI - 171 DP - 2015 May TI - A novel variant of aquaporin 3 is expressed in killifish (Fundulus heteroclitus) intestine. PG - 1-7 LID - S1532-0456(15)00021-6 [pii] LID - 10.1016/j.cbpc.2015.03.001 [doi] AB - Killifish (Fundulus heteroclitus) are euryhaline teleosts that are widely used in environmental and toxicological studies, and they are tolerant to arsenic, in part due to very low assimilation of arsenic from the environment. The mechanism of arsenic uptake by the intestine, a major route of arsenic uptake in humans is unknown. Thus, the goal of this study was to determine if aquaglyceroporins (AQPs), which transport water and other small molecules including arsenite across cell membranes, are expressed in the killifish intestine, and whether AQP expression is affected by osmotic stress. Through RT-PCR and sequence analysis of PCR amplicons, we demonstrated that the intestine expresses kfAQP3a and kfAQP3b, two previously identified variants, and also identified a novel variant of killifish AQP3 (kfAQP3c) in the intestine. The variants likely represent alternate splice forms. A BLAST search of the F. heteroclitus reference genome revealed that the AQP3 gene resides on a single locus, while an alignment of the AQP3 sequence among 384 individuals from eight population ranging from Rhode Island to North Carolina revealed that its coding sequence was remarkably conserved with no fixed polymorphism residing in the region that distinguishes these variants. We further demonstrate that the novel variant transports arsenite into HEK293T cells. Whereas kfAQP3a, which does not transport arsenite, was expressed in both freshwater (FW) and saltwater (SW) acclimated fish, kfAQP3b, an arsenic transporter, was expressed only in FW acclimated fish, and kfAQP3c was expressed only in SW acclimated fish. Thus, we have identified a novel, putative splice variant of kfAQP3, kfAQP3c, which transports arsenic and is expressed only in SW acclimated fish. CI - Copyright (c) 2015 Elsevier Inc. All rights reserved. FAU - Jung, Dawoon AU - Jung D AD - Department of Microbiology and Immunology and of Physiology, The Geisel School of Medicine at Dartmouth, Hanover, NH 03755, USA; Mt. Desert Island Biological Laboratory, Salisbury Cove, ME 04672, USA. Electronic address: Dawoon.Jung@Dartmouth.Edu. FAU - Adamo, Meredith A AU - Adamo MA AD - Mt. Desert Island Biological Laboratory, Salisbury Cove, ME 04672, USA. FAU - Lehman, Rebecca M AU - Lehman RM AD - Mt. Desert Island Biological Laboratory, Salisbury Cove, ME 04672, USA. FAU - Barnaby, Roxanna AU - Barnaby R AD - Department of Microbiology and Immunology and of Physiology, The Geisel School of Medicine at Dartmouth, Hanover, NH 03755, USA. FAU - Jackson, Craig E AU - Jackson CE AD - The School of Public and Environmental Affairs, Indiana University, Bloomington, IN 47405, USA. FAU - Jackson, Brian P AU - Jackson BP AD - Department of Earth Sciences and Chemistry, Dartmouth College, Hanover, NH 03755, USA. FAU - Shaw, Joseph R AU - Shaw JR AD - Mt. Desert Island Biological Laboratory, Salisbury Cove, ME 04672, USA; The School of Public and Environmental Affairs, Indiana University, Bloomington, IN 47405, USA; School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK. FAU - Stanton, Bruce A AU - Stanton BA AD - Department of Microbiology and Immunology and of Physiology, The Geisel School of Medicine at Dartmouth, Hanover, NH 03755, USA; Mt. Desert Island Biological Laboratory, Salisbury Cove, ME 04672, USA. LA - eng GR - P30 GM106394/GM/NIGMS NIH HHS/United States GR - P42 ES007373/ES/NIEHS NIH HHS/United States GR - P20 RR016463/RR/NCRR NIH HHS/United States GR - P20 RR-016463/RR/NCRR NIH HHS/United States GR - P20 GM103423/GM/NIGMS NIH HHS/United States GR - R01 ES019324/ES/NIEHS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20150309 PL - United States TA - Comp Biochem Physiol C Toxicol Pharmacol JT - Comparative biochemistry and physiology. Toxicology & pharmacology : CBP JID - 100959500 RN - 0 (Arsenites) RN - 0 (Fish Proteins) RN - 0 (Recombinant Proteins) RN - 158801-98-0 (Aquaporin 3) RN - N5509X556J (arsenite) SB - IM MH - Alternative Splicing MH - Animals MH - Aquaporin 3/chemistry/genetics/*metabolism MH - Arsenites/metabolism MH - Base Sequence MH - Biological Transport MH - Conserved Sequence MH - Estuaries MH - Fish Proteins/chemistry/genetics/*metabolism MH - Fundulidae/growth & development/*physiology MH - *Gene Expression Regulation MH - HEK293 Cells MH - Humans MH - Intestinal Mucosa/*metabolism MH - Maine MH - Molecular Sequence Data MH - New England MH - *Osmoregulation MH - Recombinant Proteins/chemistry/metabolism MH - Salinity MH - Sequence Alignment MH - Southeastern United States MH - *Stress, Physiological PMC - PMC4402271 MID - NIHMS670697 OTO - NOTNLM OT - Aquaglyceroporin OT - Arsenite uptake OT - Fundulus heteroclitus OT - Intestine OT - Salinity EDAT- 2015/03/15 06:00 MHDA- 2016/01/09 06:00 PMCR- 2016/05/01 CRDT- 2015/03/14 06:00 PHST- 2014/12/07 00:00 [received] PHST- 2015/03/01 00:00 [revised] PHST- 2015/03/03 00:00 [accepted] PHST- 2015/03/14 06:00 [entrez] PHST- 2015/03/15 06:00 [pubmed] PHST- 2016/01/09 06:00 [medline] PHST- 2016/05/01 00:00 [pmc-release] AID - S1532-0456(15)00021-6 [pii] AID - 10.1016/j.cbpc.2015.03.001 [doi] PST - ppublish SO - Comp Biochem Physiol C Toxicol Pharmacol. 2015 May;171:1-7. doi: 10.1016/j.cbpc.2015.03.001. Epub 2015 Mar 9.