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DEVELOPMENTAL PHYSIOLOGY AND PREGNANCY

Visualization in zebrafish larvae of Na⁺ uptake in mitochondria-rich cells whose differentiation is dependent on foxi3a

¹Department of Biological Sciences, Tokyo Institute of Technology, Yokohama; and ²Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizuoka, Japan

Submitted 19 March 2006 ; accepted in final form 25 August 2006

Uptake of Na⁺ from the environment is an indispensable strategy for the survival of freshwater fish, as they easily lose Na⁺ from the plasma to a diluted environment. Nevertheless, the location of and molecules involved in Na⁺ uptake remain poorly understood. In this study, we utilized Sodium Green, a Na⁺-dependent fluorescent reagent, to provide direct evidence that Na⁺ absorption takes place in a subset of the mitochondria-rich (MR) cells on the yolk sac surface of zebrafish larvae. Combined with immunohistochemistry, we revealed that the Na⁺-absorbing MR cells were exceptionally rich in vacuolar-type H⁺-ATPase (H⁺-ATPase) but moderately rich in Na⁺-K⁺-ATPase. We also addressed the function of foxi3a, a transcription factor that is specifically expressed in the H⁺-ATPase-rich MR cells. When foxi3a was depleted from zebrafish embryos by antisense morpholino oligonucleotide injection, differentiation of the MR cells was completely blocked and Na⁺ influx was severely reduced, indicating that MR cells are the primary sites for Na⁺ absorption. Additionally, foxi3a expression is initiated at the gastrula stage in the presumptive ectoderm; thus, we propose that foxi3a is a key gene in the control of MR cell differentiation. We also utilized a set of ion transport inhibitors to assess the molecules involved in the process and discuss the observations.

chloride cell; Sodium Green; Na⁺/H⁺ exchanger; vacuolar-type H⁺-ATPase

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