Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Sci Rep
2023 Mar 17;131:4416. doi: 10.1038/s41598-023-31503-0.
Show Gene links
Show Anatomy links
Hiat1 as a new transporter involved in ammonia regulation.
Fehsenfeld S
,
Quijada-Rodriguez AR
,
Zhouyao H
,
Durant AC
,
Donini A
,
Sachs M
,
Eck P
,
Weihrauch D
.
???displayArticle.abstract???
The orphan transporter hippocampus-abundant transcript 1 (Hiat1) was first identified in the mammalian brain. Its specific substrate specificity, however, has not been investigated to date. Here, we identified and analyzed Hiat1 in a crustacean, the green crab Carcinus maenas. Our phylogenetic analysis showed that Hiat1 protein is conserved at a considerable level between mammals and this invertebrate (ca. 78% identical and conserved amino acids). Functional expression of Carcinus maenas Hiat1 in Xenopus laevis oocytes demonstrated the capability to transport ammonia (likely NH4+) in a sodium-dependent manner. Furthermore, applying quantitative polymerase chain reaction, our results indicated a physiological role for Carcinus maenas Hiat1 in ammonia homeostasis, as mRNA abundance increased in posterior gills in response to elevated circulating hemolymph ammonia upon exposure to high environmental ammonia. Its ubiquitous mRNA expression pattern also suggests an essential role in general cellular detoxification of ammonia. Overall, our results introduce a new ubiquitously expressed ammonia transporter, consequently demanding revision of our understanding of ammonia handling in key model systems from mammalian kidneys to crustacean and fish gills.
Figure 3. Flux studies in Xenopus laevis oocytes expressing Carcinus maenas Hiat1. (A) Absolute H3-methylamine/ammonium (MA/MA+) uptake of sham-injected oocytes (white bar) and oocytes expressing Carcinus maenas Hiat1 (dark grey bar) under control conditions (medium pH 7.5, bath [total MA/MA+]â=â1 mmolâ=â10 μCi). (B) Net changes in MA/MA+ uptake (calculated as [Carcinus maenas Hiat1âsham]) at (i) pH 7.5 as referred to in panel (A), (ii) medium containing 1 mmol Lâ1 NH4Cl, and (iii) medium without Na+. Positive bars represent higher and negative bars represent lower MA/MA+ uptake of Carcinus maenas Hiat1-expressing oocytes relative to sham-injected oocytes. Asterisk indicates significant differences of MA/MA+ uptake between sham-injected and Carcinus maenas Hiat1-expressing oocytes (without sham-subtraction). Daggers indicate significant differences of MA/MA+ uptake between control conditions (pH 7.5 sham-subtracted, light grey bar) and treatments (black bars, sham subtracted) (Studentâs T-tests with Pâ<â0.01, Nâ=â20). Values are presented as meansâ±âSE. Experiments have been conducted on two different batches of oocytes (i.e., different female, different days).
Figure 4. SIET measurements of net fluxes in Xenopus laevis oocytes expressing Carcinus maenas Hiat1. (A) NH4+ uptake (positive values, bath to oocyte) and (B) H+ release (negative values, oocyte to bath) kinetics of sham-injected (dotted line, empty circles) and Carcinus maenas Hiat1-expressing (solid line, black squares) oocytes in baths with increasing NH4Cl concentration. Asterisks in (A, B) denote significant differences between sham-injected and Carcinus maenas Hiat1-expressing oocytes with regards to either NH4+ or H+ flux at the given bath [NH4Cl] (two-way ANOVA with Bonferroni multiple comparisons, Pâ<â0.05, Nâ=â5). Values are presented as meansâ±âSE. Experiments have been conducted on two different batch of oocytes (i.e., different female, different days).
Figure 1. Phylogenetic analysis of Carcinus maenas Hiat1. Shown is the Maximum likelihood consensus tree of the MUSCLE alignment of Hiat1 protein as performed with MEGAX22. Numbers beside branches represent bootstrap values (1000 replicates). Percentages indicate the level of conservation between respective species (identical + strong residues, as determined by NCBI alignment). The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. A, Arthropoda; AM, Amphibia; AV, Aves; C, Crustacea; CN, Cnidaria; E, Echinodermata; EL, Elasmobranchii; F, Fungi; M, Mammalia; N, Nematoda; P, Plantae; PL, Placozoa; PO, Porifera; TE, Teleostei, TU, Tunicata.
Figure 2. Analysis of Carcinus maenas Hiat1 mRNA abundance by quantitative PCR. (A) Organ and tissue mRNA abundance profile of Carcinus maenas Hiat1 (black bars); (B) Short-term (24 h) acclimation of green crabs to high environmental ammonia of 1 mmol L−1 (HEA). Grey bars referred to on the left y-axis in panel (B) represent mRNA abundance of Carcinus maenas Hiat1 in posterior gill #7 at 0, 6, and 24 h of HEA acclimation. Line graph referred to on the right y-axis in panel (B) represents fold change in hemolymph ammonia levels after 0, 2, 6, and 24 h of HEA acclimation. Carcinus maenas Hiat1 has been normalized to either Ribosomal gene 3 (Rbs-3, panel (A)) or elongation factor 1α (Ef1α, panel (B)). Upper case letters denote significant differences in mRNA abundance, lower case italic letters denote significant changes in hemolymph ammonia (ANOVA with Tukey’s post-hoc analysis, P > 0.05, Ntissue = 5; NHemoAmm = 6; NRNA,0 h = 6, N RNA,6 h = 7, N RNA,24 h = 8). Values are presented as means ± SE. Cer.gang., cerebral ganglion; Hypoderm., hypodermis; Ant. gland, antennal gland; Ant. gill, anterior respiratory gill #5; Post. gill, posterior osmoregulatory active gill #7.
Abdulnour-Nakhoul,
Structural determinants of NH3 and NH4+ transport by mouse Rhbg, a renal Rh glycoprotein.
2016, Pubmed,
Xenbase
Abdulnour-Nakhoul,
Structural determinants of NH3 and NH4+ transport by mouse Rhbg, a renal Rh glycoprotein.
2016,
Pubmed
,
Xenbase
Andrade,
The Amt/Mep/Rh family of ammonium transport proteins.
2007,
Pubmed
Burckhardt,
Pathways of NH3/NH4+ permeation across Xenopus laevis oocyte cell membrane.
1992,
Pubmed
,
Xenbase
Caner,
Mechanisms of ammonia and ammonium transport by rhesus-associated glycoproteins.
2015,
Pubmed
,
Xenbase
Carrisoza-Gaytán,
The hyperpolarization-activated cyclic nucleotide-gated HCN2 channel transports ammonium in the distal nephron.
2011,
Pubmed
,
Xenbase
Chasiotis,
An animal homolog of plant Mep/Amt transporters promotes ammonia excretion by the anal papillae of the disease vector mosquito Aedes aegypti.
2016,
Pubmed
Cooper,
Biochemistry and physiology of brain ammonia.
1987,
Pubmed
Donini,
Analysis of Na+, Cl-, K+, H+ and NH4+ concentration gradients adjacent to the surface of anal papillae of the mosquito Aedes aegypti: application of self-referencing ion-selective microelectrodes.
2005,
Pubmed
Doran,
Mfsd14a (Hiat1) gene disruption causes globozoospermia and infertility in male mice.
2016,
Pubmed
Durant,
Ammonium transporter expression in sperm of the disease vector Aedes aegypti mosquito influences male fertility.
2020,
Pubmed
Edgar,
MUSCLE: a multiple sequence alignment method with reduced time and space complexity.
2004,
Pubmed
Fehsenfeld,
The role of an ancestral hyperpolarization-activated cyclic nucleotide-gated K+ channel in branchial acid-base regulation in the green crab, Carcinus maenas.
2016,
Pubmed
Fehsenfeld,
Section-specific expression of acid-base and ammonia transporters in the kidney tubules of the goldfish Carassius auratus and their responses to feeding.
2018,
Pubmed
Fehsenfeld,
Differential acid-base regulation in various gills of the green crab Carcinus maenas: Effects of elevated environmental pCO2.
2013,
Pubmed
Fehsenfeld,
Effects of elevated seawater pCO(2) on gene expression patterns in the gills of the green crab, Carcinus maenas.
2011,
Pubmed
Kumar,
MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.
2018,
Pubmed
Lekholm,
Putative Membrane-Bound Transporters MFSD14A and MFSD14B Are Neuronal and Affected by Nutrient Availability.
2017,
Pubmed
Marini,
A family of ammonium transporters in Saccharomyces cerevisiae.
1997,
Pubmed
Martin,
Effects of high environmental ammonia on branchial ammonia excretion rates and tissue Rh-protein mRNA expression levels in seawater acclimated Dungeness crab Metacarcinus magister.
2011,
Pubmed
Matsuo,
Cloning of a cDNA encoding a novel sugar transporter expressed in the neonatal mouse hippocampus.
1997,
Pubmed
Méndez-Narváez,
Reproductive colonization of land by frogs: Embryos and larvae excrete urea to avoid ammonia toxicity.
2022,
Pubmed
Nawata,
Functional characterization of Rhesus glycoproteins from an ammoniotelic teleost, the rainbow trout, using oocyte expression and SIET analysis.
2010,
Pubmed
,
Xenbase
Ninnemann,
Identification of a high affinity NH4+ transporter from plants.
1994,
Pubmed
Omasits,
Protter: interactive protein feature visualization and integration with experimental proteomic data.
2014,
Pubmed
Sachs,
Characterization of two novel ammonia transporters, HIAT1α and HIAT1β, in the American Horseshoe Crab, Limulus polyphemus.
2023,
Pubmed
,
Xenbase
Si,
Identification of the role of Rh protein in ammonia excretion of the swimming crab Portunus trituberculatus.
2018,
Pubmed
Sobczak,
Endogenous transport systems in the Xenopus laevis oocyte plasma membrane.
2010,
Pubmed
,
Xenbase
Soreq,
Xenopus oocyte microinjection: from gene to protein.
1992,
Pubmed
,
Xenbase
Sreedharan,
Long evolutionary conservation and considerable tissue specificity of several atypical solute carrier transporters.
2011,
Pubmed
Towle,
Ammonium ion substitutes for K+ in ATP-dependent Na+ transport by basolateral membrane vesicles.
1987,
Pubmed
Weihrauch,
Ammonia excretion in aquatic and terrestrial crabs.
2004,
Pubmed
Weihrauch,
Active ammonia excretion across the gills of the green shore crab Carcinus maenas: participation of Na(+)/K(+)-ATPase, V-type H(+)-ATPase and functional microtubules.
2002,
Pubmed
,
Xenbase
Weiner,
Ammonia Transporters and Their Role in Acid-Base Balance.
2017,
Pubmed
Weiner,
Ammonia transport in the kidney by Rhesus glycoproteins.
2014,
Pubmed
Wright,
A new paradigm for ammonia excretion in aquatic animals: role of Rhesus (Rh) glycoproteins.
2009,
Pubmed
Zhouyao,
Characterization of two novel ammonia transporters, Hiat1a and Hiat1b, in the teleost model system Danio rerio.
2022,
Pubmed
,
Xenbase
