Why can Mozambique Tilapia Acclimate to Both Freshwater and Seawater? Insights From the Plasticity of Ionocyte Functions in the Euryhaline Teleost.

In teleost fishes, ionocytes in the gills are important osmoregulatory sites in maintaining ionic balance. During the embryonic stages before the formation of the gills, ionocytes are located in the yolk-sac membrane and body skin. In Mozambique tilapia embryos, quintuple-color immunofluorescence staining allowed us to classify ionocytes into four types: type I, showing only basolateral Na/K-ATPase (NKA) staining; type II, basolateral NKA and apical Na, Cl cotransporter 2; type III, basolateral NKA, basolateral Na, K, 2Cl cotransporter 1a (NKCC1a) and apical Na/H exchanger 3; and type IV, basolateral NKA, basolateral NKCC1a and apical cystic fibrosis transmembrane conductance regulator Cl channel.

Brood pouch evolution in pipefish and seahorse based on histological observation.

Introduction: Fishes of the Syngnathidae family are rare in having male pregnancy: males receive eggs from females and egg development occurs in the male brood pouch that diverged during evolution. The family is divided into two subfamilies: Nerophinae and Syngnathinae.

Methods: We compared histologically five types of the brood pouch in Syngnathinae: an open pouch without skinfolds (alligator pipefish); an open pouch with skinfolds (messmate pipefish); a closed pouch with skinfolds (seaweed pipefish); and closed pouches with a sac-like pouch on the tail (pot-bellied seahorse) or within a body cavity (Japanese pygmy seahorse).

Pactacin is a novel digestive enzyme in teleosts.

Generally, animals extract nutrients from food by degradation using digestive enzymes. Trypsin and chymotrypsin, one of the major digestive enzymes in vertebrates, are pancreatic proenzymes secreted into the intestines. In this investigation, we report the identification of a digestive teleost enzyme, a pancreatic astacin that we termed pactacin.

Ion uptake pathways in European sea bass Dicentrarchus labrax.

Ion uptake mechanisms are diverse in fish species, certainly linked to duplication events that have led to the presence of a multitude of paralogous genes. In fish, Na uptake involves several ion transporters expressed in different ionocyte subtypes. In the European sea bass Dicentrarchus labrax, several key transporters potentially involved in Na uptake have been investigated in seawater (SW) and following a 2 weeks freshwater (FW) acclimation.

Functional classification of gill ionocytes and spatiotemporal changes in their distribution after transfer from seawater to freshwater in Japanese seabass.

Spatiotemporal changes in branchial ionocyte distribution were investigated following transfer from seawater (SW) to freshwater (FW) in Japanese seabass. The mRNA expression levels of cystic fibrosis transmembrane conductance regulator (CFTR) and Na/K/2Cl cotransporter 1a (NKCC1a) in the gills rapidly decreased after transfer to FW, whereas Na/H exchanger 3 (NHE3) and Na/Cl cotransporter 2 (NCC2) expression were upregulated following the transfer. Using quadruple-color whole-mount immunofluorescence staining with anti-Na/K-ATPase, anti-NHE3, anti-CFTR and T4 (anti-NKCC1a/NCC2) antibodies, we classified ionocytes into one SW type and two FW types: NHE3 cell and NCC2 cell.

Morphology of brood pouch formation in the pot-bellied seahorse .

Background: The reproductive strategies of vertebrates are diverse. Seahorses (Pisces: Syngnathidae) possess the unique characteristic of male pregnancy; i.e.

Comparison of Egg Envelope Thickness in Teleosts and its Relationship to the Sites of ZP Protein Synthesis.

Teleost egg envelope generally consists of a thin outer layer and a thick inner layer. The inner layer of the Pacific herring egg envelope is further divided into distinct inner layers I and II. In our previous study, we cloned four zona pellucida (ZP) proteins (HgZPBa, HgZPBb, HgZPCa, and HgZPCb) from Pacific herring, two of which (HgZPBa and HgZPCa) were synthesized in the liver and two (HgZPBb and HgZPCb) in the ovary.

Different mechanisms of Na uptake and ammonia excretion by the gill and yolk sac epithelium of early life stage rainbow trout.

In rainbow trout, the dominant site of Na uptake () and ammonia excretion () shifts from the skin to the gills over development. Post-hatch (PH; 7 days post-hatch) larvae utilize the yolk sac skin for physiological exchange, whereas by complete yolk sac absorption (CYA; 30 days post-hatch), the gill is the dominant site. At the gills, and occur via loose Na/NH exchange, but this exchange has not been examined in the skin of larval trout.

(Uncommon) Mechanisms of Branchial Ammonia Excretion in the Common Carp (Cyprinus carpio) in Response to Environmentally Induced Metabolic Acidosis.

Freshwater fishes generally increase ammonia excretion in acidic waters. The new model of ammonia transport in freshwater fish involves an association between the Rhesus (Rh) protein Rhcg-b, the Na(+)/H(+) exchanger (NHE), and a suite of other membrane transporters. We tested the hypothesis that Rhcg-b and NHE3 together play a critical role in branchial ammonia excretion in common carp (Cyprinus carpio) chronically exposed to a low-pH environment.

An evolutionary insight into the hatching strategies of pipefish and seahorse embryos.

Syngnathiform fishes carry their eggs in a brood structure found in males. The brood structure differs from species to species: seahorses carry eggs within enclosed brood pouch, messmate pipefish carry eggs in the semi-brood pouch, and alligator pipefish carry eggs in the egg compartment on abdomen. These egg protection strategies were established during syngnathiform evolution.

Diverse mechanisms for body fluid regulation in teleost fishes.

Teleost fishes are the major group of ray-finned fishes and represent more than one-half of the total number of vertebrate species. They have experienced in their evolution an additional third-round whole genome duplication just after the divergence of their lineage, which endowed them with an extra adaptability to invade various aquatic habitats. Thus their physiology is also extremely diverse compared with other vertebrate groups as exemplified by the many patterns of body fluid regulation or osmoregulation.

New insights into gill ionocyte and ion transporter function in euryhaline and diadromous fish.

Teleost fishes are able to acclimatize to seawater by secreting excess NaCl by means of specialized "ionocytes" in the gill epithelium. Antibodies against Na(+)/K(+)-ATPase (NKA) have been used since 1996 as a marker for identifying branchial ionocytes. Immunohistochemistry of NKA by itself and in combination with Na(+)/K(+)/2Cl(-) cotransporter and CFTR Cl(-) channel provided convincing evidence that ionocytes are functional during seawater acclimation, and also revealed morphological variations in ionocytes among teleost species.

Intron-loss evolution of hatching enzyme genes in Teleostei.

Background: Hatching enzyme, belonging to the astacin metallo-protease family, digests egg envelope at embryo hatching. Orthologous genes of the enzyme are found in all vertebrate genomes. Recently, we found that exon-intron structures of the genes were conserved among tetrapods, while the genes of teleosts frequently lost their introns.

Morphological and functional classification of ion-absorbing mitochondria-rich cells in the gills of Mozambique tilapia.

To clarify ion-absorbing functions and molecular mechanisms of mitochondria-rich (MR) cells, Mozambique tilapia (Oreochromis mossambicus) were acclimated to artificial freshwaters with normal or lowered Na+ and/or Cl- concentration: (1) normal Na+/normal Cl- (control); (2) normal Na+/low Cl-; (3) low Na+/normal Cl-; and (4) low Na+/low Cl-. Scanning electron microscopy (SEM) revealed that concave and convex apical surfaces of MR cells predominantly developed in low Na+ and low Cl- waters, respectively, whereas small apical pits predominated in control conditions. Expression of Na+/H+ exchanger-3 (NHE3) mRNA in the gills was increased in low Na+ waters (low Na+/normal Cl- and low Na+/low Cl-, whereas that of Na+/Cl- cotransporter (NCC) expression was upregulated in low Cl-, but not in low Na+/low Cl-.

Role of SLC12A10.2, a Na-Cl cotransporter-like protein, in a Cl uptake mechanism in zebrafish (Danio rerio).

The thiazide-sensitive Na(+)-Cl(-) cotransporter (NCC), a member of the SLC12 family, is mainly expressed in the apical membrane of the mammalian distal convoluted tubule (DCT) cells, is responsible for cotransporting Na(+) and Cl(-) from the lumen into DCT cells and plays a major role in the mammalian renal NaCl reabsorption. The NCC has also been reported in fish, but the functional role in fish ion regulation is yet unclear. The present study used zebrafish as an in vivo model to test the hypothesis of whether the NCC plays a role in Na(+) and/or Cl(-) uptake mechanisms.

Different hatching strategies in embryos of two species, pacific herring Clupea pallasii and Japanese anchovy Engraulis japonicus, that belong to the same order Clupeiformes, and their environmental adaptation.

Pacific herring Clupea pallasii and Japanese anchovy Engraulis japonicus, which belong to the same order Clupeiformes, spawn different types of eggs: demersal adherent eggs and pelagic eggs, respectively. We cloned three cDNAs for Pacific herring hatching enzyme and five for Japanese anchovy. Each of them was divided into two groups (group A and B) by phylogenetic analysis.

Evidence for an apical Na-Cl cotransporter involved in ion uptake in a teleost fish.

Cation-chloride cotransporters, such as the Na(+)/K(+)/2Cl(-) cotransporter (NKCC) and Na(+)/Cl(-) cotransporter (NCC), are localized to the apical or basolateral plasma membranes of epithelial cells and are involved in active ion absorption or secretion. The objectives of this study were to clone and identify ;freshwater-type' and ;seawater-type' cation-chloride cotransporters of euryhaline Mozambique tilapia (Oreochromis mossambicus) and to determine their intracellular localization patterns within mitochondria-rich cells (MRCs). From tilapia gills, we cloned four full-length cDNAs homologous to human cation-chloride cotransporters and designated them as tilapia NKCC1a, NKCC1b, NKCC2 and NCC.

Gene expression and morphological localization of NHE3, NCC and NKCC1a in branchial mitochondria-rich cells of Mozambique tilapia (Oreochromis mossambicus) acclimated to a wide range of salinities.

We explored molecular and morphological alteration in gill mitochondria-rich (MR) cells of Mozambique tilapia, Oreochromis mossambicus, acclimated to deionized freshwater (DFW), freshwater (FW), 1/3-diluted seawater (1/3 SW) and seawater (SW). Scanning electron microscopic observations revealed that the apical membrane of MR cells appeared as a flat or slightly projecting disk in DFW and FW, being larger in DFW than in FW. In contrast, the apical membrane typically formed a pit structure in 1/3 SW and SW.

Hatching enzyme of the ovoviviparous black rockfish Sebastes schlegelii- environmental adaptation of the hatching enzyme and evolutionary aspects of formation of the pseudogene.

The hatching enzyme of oviparous euteleostean fishes consists of two metalloproteases: high choriolytic enzyme (HCE) and low choriolytic enzyme (LCE). They cooperatively digest the egg envelope (chorion) at the time of embryo hatching. In the present study, we investigated the hatching of embryos of the ovoviviparous black rockfish Sebastes schlegelii.

Variation in salinity tolerance, gill Na+/K+-ATPase, Na+/K+/2Cl- cotransporter and mitochondria-rich cell distribution in three salmonids Salvelinus namaycush, Salvelinus fontinalis and Salmo salar.

We compared seawater tolerance, gill Na(+)/K(+)-ATPase and Na(+)/K(+)/2Cl(-) cotransporter (NKCC) abundance, and mitochondria-rich cell (MRC) morphology of three salmonids, lake trout Salvelinus namaycush, brook trout Salvelinus fontinalis and Atlantic salmon Salmo salar. They were transferred directly from 0 p.p.

Analysis of the exon-intron structures of fish, amphibian, bird and mammalian hatching enzyme genes, with special reference to the intron loss evolution of hatching enzyme genes in Teleostei.

Using gene cloning and in silico cloning, we analyzed the structures of hatching enzyme gene orthologs of vertebrates. Comparison led to a hypothesis that hatching enzyme genes of Japanese eel conserve an ancestral structure of the genes of fishes, amphibians, birds and mammals. However, the exon-intron structure of the genes was different from species to species in Teleostei: Japanese eel hatching enzyme genes were 9-exon-8-intron genes, and zebrafish genes were 5-exon-4-intron genes.

Evolution of teleostean hatching enzyme genes and their paralogous genes.

We isolated genes for hatching enzymes and their paralogs having two cysteine residues at their N-terminal regions in addition to four cysteines conserved in all the astacin family proteases. Genes for such six-cysteine-containing astacin proteases (C6AST) were searched out in the medaka genome database. Five genes for MC6AST1 to 5 were found in addition to embryo-specific hatching enzyme genes.

Liver-type fatty acid-binding protein attenuates renal injury induced by unilateral ureteral obstruction.

Liver-type fatty-acid-binding protein (L-FABP), which has high affinity for long-chain fatty acid oxidation products, may be an effective endogenous antioxidant. To examine the role of L-FABP in tubulointerstitial damage, we used a unilateral ureteral obstruction (UUO) model. We established human L-FABP (hL-FABP) gene transgenic (Tg) mice and compared the tubulointerstitial pathology of the Tg mice (n = 23) with that of the wild-type (WT) mice (n = 23).