Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice

Kenta Wada, Yoshibumi Matsushima, Tomoki Tada, Sayaka Hasegawa, Yo Obara, Yasuhiro Yoshizawa, Gou Takahashi, Hiroshi Hiai, Midori Shimanuki, Sari Suzuki, Junichi Saitou, Naoki Yamamoto, Masumi Ichikawa, Kei Watanabe, Yoshiaki Kikkawa

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Microphthalmia is a severe ocular disorder, and this condition is typically caused by mutations in transcription factors that are involved in eye development. Mice carrying mutations in these transcription factors would be useful tools for defining the mechanisms underlying developmental eye disorders. We discovered a new spontaneous recessive microphthalmos mouse mutant in the Japanese wild-derived inbred strain KOR1/Stm. The homozygous mutant mice were histologically characterized as microphthalmic by the absence of crystallin in the lens, a condition referred to as aphakia. By positional cloning, we identified the nonsense mutation c.444C>A outside the genomic region that encodes the homeodomain of the paired-like homeodomain transcription factor 3 gene (Pitx3) as the mutation responsible for the microphthalmia and aphakia. We examined Pitx3 mRNA expression of mutant mice during embryonic stages using RT-PCR and found that the expression levels are higher than in wild-type mice. Pitx3 over-expression in the lens during developmental stages was also confirmed at the protein level in the microphthalmos mutants via immunohistochemical analyses. Although lens fiber differentiation was not observed in the mutants, strong PITX3 protein signals were observed in the lens vesicles of the mutant lens. Thus, we speculated that abnormal PITX3, which lacks the C-terminus (including the OAR domain) as a result of the nonsense mutation, is expressed in mutant lenses. We showed that the expression of the downstream genes Foxe3, Prox1, and Mip was altered because of the Pitx3 mutation, with large reductions in the lens vesicles in the mutants. Similar profiles were observed by immunohistochemical analysis of these proteins. The expression profiles of crystallins were also altered in the mutants. Therefore, we speculated that the microphthalmos/aphakia in this mutant is caused by the expression of truncated PITX3, resulting in the abnormal expression of downstream targets and lens fiber proteins.

Original languageEnglish
Article numbere111432
JournalPloS one
Volume9
Issue number10
DOIs
Publication statusPublished - 27-10-2014

Fingerprint

Aphakia
Microphthalmos
Lens
Lenses
mutants
mice
Crystallins
Mutation
Nonsense Codon
nonsense mutation
crystallins
mutation
transcription factors
eyes
Transcription Factors
Proteins
Genes
proteins
Fibers
Cloning

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Wada, K., Matsushima, Y., Tada, T., Hasegawa, S., Obara, Y., Yoshizawa, Y., ... Kikkawa, Y. (2014). Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice. PloS one, 9(10), [e111432]. https://doi.org/10.1371/journal.pone.0111432
Wada, Kenta ; Matsushima, Yoshibumi ; Tada, Tomoki ; Hasegawa, Sayaka ; Obara, Yo ; Yoshizawa, Yasuhiro ; Takahashi, Gou ; Hiai, Hiroshi ; Shimanuki, Midori ; Suzuki, Sari ; Saitou, Junichi ; Yamamoto, Naoki ; Ichikawa, Masumi ; Watanabe, Kei ; Kikkawa, Yoshiaki. / Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice. In: PloS one. 2014 ; Vol. 9, No. 10.
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abstract = "Microphthalmia is a severe ocular disorder, and this condition is typically caused by mutations in transcription factors that are involved in eye development. Mice carrying mutations in these transcription factors would be useful tools for defining the mechanisms underlying developmental eye disorders. We discovered a new spontaneous recessive microphthalmos mouse mutant in the Japanese wild-derived inbred strain KOR1/Stm. The homozygous mutant mice were histologically characterized as microphthalmic by the absence of crystallin in the lens, a condition referred to as aphakia. By positional cloning, we identified the nonsense mutation c.444C>A outside the genomic region that encodes the homeodomain of the paired-like homeodomain transcription factor 3 gene (Pitx3) as the mutation responsible for the microphthalmia and aphakia. We examined Pitx3 mRNA expression of mutant mice during embryonic stages using RT-PCR and found that the expression levels are higher than in wild-type mice. Pitx3 over-expression in the lens during developmental stages was also confirmed at the protein level in the microphthalmos mutants via immunohistochemical analyses. Although lens fiber differentiation was not observed in the mutants, strong PITX3 protein signals were observed in the lens vesicles of the mutant lens. Thus, we speculated that abnormal PITX3, which lacks the C-terminus (including the OAR domain) as a result of the nonsense mutation, is expressed in mutant lenses. We showed that the expression of the downstream genes Foxe3, Prox1, and Mip was altered because of the Pitx3 mutation, with large reductions in the lens vesicles in the mutants. Similar profiles were observed by immunohistochemical analysis of these proteins. The expression profiles of crystallins were also altered in the mutants. Therefore, we speculated that the microphthalmos/aphakia in this mutant is caused by the expression of truncated PITX3, resulting in the abnormal expression of downstream targets and lens fiber proteins.",
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Wada, K, Matsushima, Y, Tada, T, Hasegawa, S, Obara, Y, Yoshizawa, Y, Takahashi, G, Hiai, H, Shimanuki, M, Suzuki, S, Saitou, J, Yamamoto, N, Ichikawa, M, Watanabe, K & Kikkawa, Y 2014, 'Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice', PloS one, vol. 9, no. 10, e111432. https://doi.org/10.1371/journal.pone.0111432

Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice. / Wada, Kenta; Matsushima, Yoshibumi; Tada, Tomoki; Hasegawa, Sayaka; Obara, Yo; Yoshizawa, Yasuhiro; Takahashi, Gou; Hiai, Hiroshi; Shimanuki, Midori; Suzuki, Sari; Saitou, Junichi; Yamamoto, Naoki; Ichikawa, Masumi; Watanabe, Kei; Kikkawa, Yoshiaki.

In: PloS one, Vol. 9, No. 10, e111432, 27.10.2014.

Research output: Contribution to journalArticle

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AU - Wada, Kenta

AU - Matsushima, Yoshibumi

AU - Tada, Tomoki

AU - Hasegawa, Sayaka

AU - Obara, Yo

AU - Yoshizawa, Yasuhiro

AU - Takahashi, Gou

AU - Hiai, Hiroshi

AU - Shimanuki, Midori

AU - Suzuki, Sari

AU - Saitou, Junichi

AU - Yamamoto, Naoki

AU - Ichikawa, Masumi

AU - Watanabe, Kei

AU - Kikkawa, Yoshiaki

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N2 - Microphthalmia is a severe ocular disorder, and this condition is typically caused by mutations in transcription factors that are involved in eye development. Mice carrying mutations in these transcription factors would be useful tools for defining the mechanisms underlying developmental eye disorders. We discovered a new spontaneous recessive microphthalmos mouse mutant in the Japanese wild-derived inbred strain KOR1/Stm. The homozygous mutant mice were histologically characterized as microphthalmic by the absence of crystallin in the lens, a condition referred to as aphakia. By positional cloning, we identified the nonsense mutation c.444C>A outside the genomic region that encodes the homeodomain of the paired-like homeodomain transcription factor 3 gene (Pitx3) as the mutation responsible for the microphthalmia and aphakia. We examined Pitx3 mRNA expression of mutant mice during embryonic stages using RT-PCR and found that the expression levels are higher than in wild-type mice. Pitx3 over-expression in the lens during developmental stages was also confirmed at the protein level in the microphthalmos mutants via immunohistochemical analyses. Although lens fiber differentiation was not observed in the mutants, strong PITX3 protein signals were observed in the lens vesicles of the mutant lens. Thus, we speculated that abnormal PITX3, which lacks the C-terminus (including the OAR domain) as a result of the nonsense mutation, is expressed in mutant lenses. We showed that the expression of the downstream genes Foxe3, Prox1, and Mip was altered because of the Pitx3 mutation, with large reductions in the lens vesicles in the mutants. Similar profiles were observed by immunohistochemical analysis of these proteins. The expression profiles of crystallins were also altered in the mutants. Therefore, we speculated that the microphthalmos/aphakia in this mutant is caused by the expression of truncated PITX3, resulting in the abnormal expression of downstream targets and lens fiber proteins.

AB - Microphthalmia is a severe ocular disorder, and this condition is typically caused by mutations in transcription factors that are involved in eye development. Mice carrying mutations in these transcription factors would be useful tools for defining the mechanisms underlying developmental eye disorders. We discovered a new spontaneous recessive microphthalmos mouse mutant in the Japanese wild-derived inbred strain KOR1/Stm. The homozygous mutant mice were histologically characterized as microphthalmic by the absence of crystallin in the lens, a condition referred to as aphakia. By positional cloning, we identified the nonsense mutation c.444C>A outside the genomic region that encodes the homeodomain of the paired-like homeodomain transcription factor 3 gene (Pitx3) as the mutation responsible for the microphthalmia and aphakia. We examined Pitx3 mRNA expression of mutant mice during embryonic stages using RT-PCR and found that the expression levels are higher than in wild-type mice. Pitx3 over-expression in the lens during developmental stages was also confirmed at the protein level in the microphthalmos mutants via immunohistochemical analyses. Although lens fiber differentiation was not observed in the mutants, strong PITX3 protein signals were observed in the lens vesicles of the mutant lens. Thus, we speculated that abnormal PITX3, which lacks the C-terminus (including the OAR domain) as a result of the nonsense mutation, is expressed in mutant lenses. We showed that the expression of the downstream genes Foxe3, Prox1, and Mip was altered because of the Pitx3 mutation, with large reductions in the lens vesicles in the mutants. Similar profiles were observed by immunohistochemical analysis of these proteins. The expression profiles of crystallins were also altered in the mutants. Therefore, we speculated that the microphthalmos/aphakia in this mutant is caused by the expression of truncated PITX3, resulting in the abnormal expression of downstream targets and lens fiber proteins.

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Wada K, Matsushima Y, Tada T, Hasegawa S, Obara Y, Yoshizawa Y et al. Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice. PloS one. 2014 Oct 27;9(10). e111432. https://doi.org/10.1371/journal.pone.0111432