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Ferritin H subunit gene is specifically expressed in melanophore precursor-derived white pigment cells in which reflecting platelets are formed from stage II melanosomes in the periodic albino mutant of Xenopus laevis.

Fukuzawa T - Cell Tissue Res. (2015)

Bottom Line: Here, white pigment cells in the mutant regenerating tail have been compared with melanophores in the wild type regenerating tail in the presence of phenylthiourea (PTU), which inhibits melanosome maturation in melanophores but does not affect reflecting platelet formation in white pigment cells.Among cDNA fragments examined so far, the ferritin H subunit gene is specifically expressed in white pigment cells, but not in melanophores.Pigment organellogenesis and specific gene expression in white pigment cells are also discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Keio University, Hiyoshi 4-1-1, Kohoku-ku, Yokohama, 223-8521, Japan, fukuzawa@fbc.keio.ac.jp.

ABSTRACT
"White pigment cells" are derived from melanophore precursors and contain both melanophore-specific and iridophore-specific pigment organelles. Whereas melanophores differentiate in the wild type regenerating tail, white pigment cells appear in the regenerating tail in the periodic albino mutant (a(p)/a(p)) of Xenopus laevis. The localization and density of white pigment cells in the mutant regenerating tail are similar to those of melanophores in the wild type regenerating tail. Here, white pigment cells in the mutant regenerating tail have been compared with melanophores in the wild type regenerating tail in the presence of phenylthiourea (PTU), which inhibits melanosome maturation in melanophores but does not affect reflecting platelet formation in white pigment cells. Ultrastructural analysis shows that reflecting platelet formation in white pigment cells is different from that in iridophores. Reflecting platelets in iridophores are formed from spherical vesicles with electron-dense material, whereas they are formed from stage II melanosomes characteristic of melanophore precursors in white pigment cells. Ultrastructural features of pigment organelles, except reflecting platelets, are similar between mutant melanophores and white pigment cells. In an attempt to identify specific genes in white pigment cells, a subtracted cDNA library enriched for mutant cDNAs has been prepared. Subtracted cDNA fragments have been cloned and selected by whole mount in situ hybridization. Among cDNA fragments examined so far, the ferritin H subunit gene is specifically expressed in white pigment cells, but not in melanophores. Pigment organellogenesis and specific gene expression in white pigment cells are also discussed.

No MeSH data available.


Related in: MedlinePlus

Spatial expression of the ferritin H subunit mRNA in the middle region of the tail in the wild type (a, b) and the mutant (c, d) at stage 48. Whole mount in situ hybridization (WISH) was performed by using sense (a, c) or antisense (b, d) digoxigenin (DIG)-labeled RNA probes. Tadpoles were bleached to remove melanin before hybridization in this experiment. With a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of both the wild type (a) and the mutant (c) in the negative control. Use of an antisense probe in WISH detected strong staining in the lateral lines (arrowheads) of both the wild type (b) and the mutant (d). In addition, specific expression of the ferritin H subunit mRNA was detected in white pigment cells (d, large arrows), which were present around the dorsal side of the spinal cord (sc) in the mutant (nc notochord). Although melanophores were present around the dorsal side of the spinal cord in the wild type, no staining was observed in melanophores (b). Note that staining was also detected in some epidermal cells (small arrows) in both the wild type (b) and the mutant (d). Bar 100 μm
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Fig6: Spatial expression of the ferritin H subunit mRNA in the middle region of the tail in the wild type (a, b) and the mutant (c, d) at stage 48. Whole mount in situ hybridization (WISH) was performed by using sense (a, c) or antisense (b, d) digoxigenin (DIG)-labeled RNA probes. Tadpoles were bleached to remove melanin before hybridization in this experiment. With a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of both the wild type (a) and the mutant (c) in the negative control. Use of an antisense probe in WISH detected strong staining in the lateral lines (arrowheads) of both the wild type (b) and the mutant (d). In addition, specific expression of the ferritin H subunit mRNA was detected in white pigment cells (d, large arrows), which were present around the dorsal side of the spinal cord (sc) in the mutant (nc notochord). Although melanophores were present around the dorsal side of the spinal cord in the wild type, no staining was observed in melanophores (b). Note that staining was also detected in some epidermal cells (small arrows) in both the wild type (b) and the mutant (d). Bar 100 μm

Mentions: WISH was performed in wild type and mutant tadpoles at stage 48 to examine the expression of the ferritin H subunit gene. Figure 6 shows the result of WISH in the middle region of the tail in which melanin was removed by bleaching before hybridization. In the negative control with a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of either the wild type (Fig. 6a) or the mutant (Fig. 6c). Hybridization with an antisense probe of the ferritin H subunit mRNA provided a clear hybridization signal (Fig. 6b, d). Strong staining was detected in the lateral lines of both the wild type and the mutant (Fig. 6b, d arrowheads). This result parallels the report of the ferritin H subunit gene being expressed in the accessory cells in the zebrafish lateral line (Behra et al. 2012). Some epidermal cells also expressed the ferritin H subunit mRNA in both the wild type and the mutant (Fig. 6b, d, small arrows). Specific expression of the ferritin H subunit mRNA was detected in white pigment cells, which were present around the dorsal side of the mutant spinal cord (Fig. 6d, large arrows). Although the localization of melanophores in the wild type was similar to that of white pigment cells in the mutant, no staining was observed in melanophores (Fig. 6b).Fig. 6


Ferritin H subunit gene is specifically expressed in melanophore precursor-derived white pigment cells in which reflecting platelets are formed from stage II melanosomes in the periodic albino mutant of Xenopus laevis.

Fukuzawa T - Cell Tissue Res. (2015)

Spatial expression of the ferritin H subunit mRNA in the middle region of the tail in the wild type (a, b) and the mutant (c, d) at stage 48. Whole mount in situ hybridization (WISH) was performed by using sense (a, c) or antisense (b, d) digoxigenin (DIG)-labeled RNA probes. Tadpoles were bleached to remove melanin before hybridization in this experiment. With a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of both the wild type (a) and the mutant (c) in the negative control. Use of an antisense probe in WISH detected strong staining in the lateral lines (arrowheads) of both the wild type (b) and the mutant (d). In addition, specific expression of the ferritin H subunit mRNA was detected in white pigment cells (d, large arrows), which were present around the dorsal side of the spinal cord (sc) in the mutant (nc notochord). Although melanophores were present around the dorsal side of the spinal cord in the wild type, no staining was observed in melanophores (b). Note that staining was also detected in some epidermal cells (small arrows) in both the wild type (b) and the mutant (d). Bar 100 μm
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Related In: Results  -  Collection

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Fig6: Spatial expression of the ferritin H subunit mRNA in the middle region of the tail in the wild type (a, b) and the mutant (c, d) at stage 48. Whole mount in situ hybridization (WISH) was performed by using sense (a, c) or antisense (b, d) digoxigenin (DIG)-labeled RNA probes. Tadpoles were bleached to remove melanin before hybridization in this experiment. With a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of both the wild type (a) and the mutant (c) in the negative control. Use of an antisense probe in WISH detected strong staining in the lateral lines (arrowheads) of both the wild type (b) and the mutant (d). In addition, specific expression of the ferritin H subunit mRNA was detected in white pigment cells (d, large arrows), which were present around the dorsal side of the spinal cord (sc) in the mutant (nc notochord). Although melanophores were present around the dorsal side of the spinal cord in the wild type, no staining was observed in melanophores (b). Note that staining was also detected in some epidermal cells (small arrows) in both the wild type (b) and the mutant (d). Bar 100 μm
Mentions: WISH was performed in wild type and mutant tadpoles at stage 48 to examine the expression of the ferritin H subunit gene. Figure 6 shows the result of WISH in the middle region of the tail in which melanin was removed by bleaching before hybridization. In the negative control with a sense probe of the ferritin H subunit mRNA, no staining was observed in the tail of either the wild type (Fig. 6a) or the mutant (Fig. 6c). Hybridization with an antisense probe of the ferritin H subunit mRNA provided a clear hybridization signal (Fig. 6b, d). Strong staining was detected in the lateral lines of both the wild type and the mutant (Fig. 6b, d arrowheads). This result parallels the report of the ferritin H subunit gene being expressed in the accessory cells in the zebrafish lateral line (Behra et al. 2012). Some epidermal cells also expressed the ferritin H subunit mRNA in both the wild type and the mutant (Fig. 6b, d, small arrows). Specific expression of the ferritin H subunit mRNA was detected in white pigment cells, which were present around the dorsal side of the mutant spinal cord (Fig. 6d, large arrows). Although the localization of melanophores in the wild type was similar to that of white pigment cells in the mutant, no staining was observed in melanophores (Fig. 6b).Fig. 6

Bottom Line: Here, white pigment cells in the mutant regenerating tail have been compared with melanophores in the wild type regenerating tail in the presence of phenylthiourea (PTU), which inhibits melanosome maturation in melanophores but does not affect reflecting platelet formation in white pigment cells.Among cDNA fragments examined so far, the ferritin H subunit gene is specifically expressed in white pigment cells, but not in melanophores.Pigment organellogenesis and specific gene expression in white pigment cells are also discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Keio University, Hiyoshi 4-1-1, Kohoku-ku, Yokohama, 223-8521, Japan, fukuzawa@fbc.keio.ac.jp.

ABSTRACT
"White pigment cells" are derived from melanophore precursors and contain both melanophore-specific and iridophore-specific pigment organelles. Whereas melanophores differentiate in the wild type regenerating tail, white pigment cells appear in the regenerating tail in the periodic albino mutant (a(p)/a(p)) of Xenopus laevis. The localization and density of white pigment cells in the mutant regenerating tail are similar to those of melanophores in the wild type regenerating tail. Here, white pigment cells in the mutant regenerating tail have been compared with melanophores in the wild type regenerating tail in the presence of phenylthiourea (PTU), which inhibits melanosome maturation in melanophores but does not affect reflecting platelet formation in white pigment cells. Ultrastructural analysis shows that reflecting platelet formation in white pigment cells is different from that in iridophores. Reflecting platelets in iridophores are formed from spherical vesicles with electron-dense material, whereas they are formed from stage II melanosomes characteristic of melanophore precursors in white pigment cells. Ultrastructural features of pigment organelles, except reflecting platelets, are similar between mutant melanophores and white pigment cells. In an attempt to identify specific genes in white pigment cells, a subtracted cDNA library enriched for mutant cDNAs has been prepared. Subtracted cDNA fragments have been cloned and selected by whole mount in situ hybridization. Among cDNA fragments examined so far, the ferritin H subunit gene is specifically expressed in white pigment cells, but not in melanophores. Pigment organellogenesis and specific gene expression in white pigment cells are also discussed.

No MeSH data available.


Related in: MedlinePlus