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Isotocin controls ion regulation through regulating ionocyte progenitor differentiation and proliferation.

Chou MY, Hung JC, Wu LC, Hwang SP, Hwang PP - Cell. Mol. Life Sci. (2010)

Bottom Line: Double-deionized water treatment for 24 h significantly stimulated isotocin mRNA expression in zebrafish embryos.Whole-body Cl(-), Ca(2+), and Na(+) contents, mRNA expressions of ion transporters and ionocyte-differentiation related transcription factors, and the number of skin ionocytes decreased in isotocin morphants.Taken together, isotocin stimulates the proliferation of epidermal stem cells and differentiation of ionocyte progenitors by regulating the P63 and Foxi3a transcription factors, consequently enhancing the functional activities of ionocytes.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, Taiwan.

ABSTRACT
The present study using zebrafish as a model explores the role of isotocin, a homolog of oxytocin, in controlling ion regulatory mechanisms. Double-deionized water treatment for 24 h significantly stimulated isotocin mRNA expression in zebrafish embryos. Whole-body Cl(-), Ca(2+), and Na(+) contents, mRNA expressions of ion transporters and ionocyte-differentiation related transcription factors, and the number of skin ionocytes decreased in isotocin morphants. In contrast, overexpression of isotocin caused an increase in ionocyte numbers. Isotocin morpholino caused significant suppression of foxi3a mRNA expression, while isotocin cRNA stimulated foxi3a mRNA expressions at the tail-bud stage of zebrafish embryos. The density of P63 (an epidermal stem cell marker)-positive cells was downregulated by isotocin morpholinos and was upregulated by isotocin cRNA. Taken together, isotocin stimulates the proliferation of epidermal stem cells and differentiation of ionocyte progenitors by regulating the P63 and Foxi3a transcription factors, consequently enhancing the functional activities of ionocytes.

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Effects of knockdown or overexpression of itnp on P63 expression in zebrafish embryos. One to two-cell stage embryos were injected with a mismatched-MO (Mis MO) (1 ng/embryo) (a), itnp MO (1 ng/embryo) (b), and isotocin cRNA (1 ng/embryo) (c), respectively, and P63+ cells in yolk-sac area were detected by immunocytochemistry at 72 hpf. d The density of P63+ cells was lower in itnp morphants but had increased in itnp cRNA-injected embryos. Mean ± SD (n = 6). Different lowercase letters indicate significant differences (one-way ANOVA, Tukey’s pair-wise comparison). Scale bar 50 μm
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Fig6: Effects of knockdown or overexpression of itnp on P63 expression in zebrafish embryos. One to two-cell stage embryos were injected with a mismatched-MO (Mis MO) (1 ng/embryo) (a), itnp MO (1 ng/embryo) (b), and isotocin cRNA (1 ng/embryo) (c), respectively, and P63+ cells in yolk-sac area were detected by immunocytochemistry at 72 hpf. d The density of P63+ cells was lower in itnp morphants but had increased in itnp cRNA-injected embryos. Mean ± SD (n = 6). Different lowercase letters indicate significant differences (one-way ANOVA, Tukey’s pair-wise comparison). Scale bar 50 μm

Mentions: The loss- and gain-of-function approaches were used to further examine if the control pathways of isotocin in ionocyte progenitor differentiation are mediated by regulating epidermal stem cells. P63 is a marker of epithelial stem cells, which were demonstrated to differentiate into skin ionocytes and keratinocytes in zebrafish [35, 36]. To test whether isotocin regulates the number of epidermal stem cells in zebrafish embryos, P63 staining was performed. The density of P63+ cells decreased in itnp morphants and, on the contrary, increased in itnp ectopically expressed embryos (Fig. 6). No difference was found between wild-type and mismatched MO-injected embryos (data not shown).Fig. 6


Isotocin controls ion regulation through regulating ionocyte progenitor differentiation and proliferation.

Chou MY, Hung JC, Wu LC, Hwang SP, Hwang PP - Cell. Mol. Life Sci. (2010)

Effects of knockdown or overexpression of itnp on P63 expression in zebrafish embryos. One to two-cell stage embryos were injected with a mismatched-MO (Mis MO) (1 ng/embryo) (a), itnp MO (1 ng/embryo) (b), and isotocin cRNA (1 ng/embryo) (c), respectively, and P63+ cells in yolk-sac area were detected by immunocytochemistry at 72 hpf. d The density of P63+ cells was lower in itnp morphants but had increased in itnp cRNA-injected embryos. Mean ± SD (n = 6). Different lowercase letters indicate significant differences (one-way ANOVA, Tukey’s pair-wise comparison). Scale bar 50 μm
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3142547&req=5

Fig6: Effects of knockdown or overexpression of itnp on P63 expression in zebrafish embryos. One to two-cell stage embryos were injected with a mismatched-MO (Mis MO) (1 ng/embryo) (a), itnp MO (1 ng/embryo) (b), and isotocin cRNA (1 ng/embryo) (c), respectively, and P63+ cells in yolk-sac area were detected by immunocytochemistry at 72 hpf. d The density of P63+ cells was lower in itnp morphants but had increased in itnp cRNA-injected embryos. Mean ± SD (n = 6). Different lowercase letters indicate significant differences (one-way ANOVA, Tukey’s pair-wise comparison). Scale bar 50 μm
Mentions: The loss- and gain-of-function approaches were used to further examine if the control pathways of isotocin in ionocyte progenitor differentiation are mediated by regulating epidermal stem cells. P63 is a marker of epithelial stem cells, which were demonstrated to differentiate into skin ionocytes and keratinocytes in zebrafish [35, 36]. To test whether isotocin regulates the number of epidermal stem cells in zebrafish embryos, P63 staining was performed. The density of P63+ cells decreased in itnp morphants and, on the contrary, increased in itnp ectopically expressed embryos (Fig. 6). No difference was found between wild-type and mismatched MO-injected embryos (data not shown).Fig. 6

Bottom Line: Double-deionized water treatment for 24 h significantly stimulated isotocin mRNA expression in zebrafish embryos.Whole-body Cl(-), Ca(2+), and Na(+) contents, mRNA expressions of ion transporters and ionocyte-differentiation related transcription factors, and the number of skin ionocytes decreased in isotocin morphants.Taken together, isotocin stimulates the proliferation of epidermal stem cells and differentiation of ionocyte progenitors by regulating the P63 and Foxi3a transcription factors, consequently enhancing the functional activities of ionocytes.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, Taiwan.

ABSTRACT
The present study using zebrafish as a model explores the role of isotocin, a homolog of oxytocin, in controlling ion regulatory mechanisms. Double-deionized water treatment for 24 h significantly stimulated isotocin mRNA expression in zebrafish embryos. Whole-body Cl(-), Ca(2+), and Na(+) contents, mRNA expressions of ion transporters and ionocyte-differentiation related transcription factors, and the number of skin ionocytes decreased in isotocin morphants. In contrast, overexpression of isotocin caused an increase in ionocyte numbers. Isotocin morpholino caused significant suppression of foxi3a mRNA expression, while isotocin cRNA stimulated foxi3a mRNA expressions at the tail-bud stage of zebrafish embryos. The density of P63 (an epidermal stem cell marker)-positive cells was downregulated by isotocin morpholinos and was upregulated by isotocin cRNA. Taken together, isotocin stimulates the proliferation of epidermal stem cells and differentiation of ionocyte progenitors by regulating the P63 and Foxi3a transcription factors, consequently enhancing the functional activities of ionocytes.

Show MeSH
Related in: MedlinePlus