Lens-preferred activity of chicken δ1- and δ2-crystallin enhancers in transgenic mice and evidence for retinoic acid-responsive regulation of the δ1-crystallin gene

Xuan Li, Ales Cvekl, Steven Bassnett, Joram Piatigorsky

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

There are two tandemly linked δ-crystallin genes (5' δ-δ2 3') in the chicken, with the δ1-crystallin gene being expressed much more highly (50- 100-fold) in the embryonic lens than the δ2-crystallin gene. Previous transfection experiments have shown that a lens preferred enhancer exists in the third intron of each chicken δ-crystallin gene. In the present investigation we have used transgenic mice to establish that both the chicken δ1- and δ2-crystallin enhancers are preferentially active in the mouse lens in combination with their homologous promoter and the chloramphenicol acetyltransferase (CAT) reporter gene. The promoter/CAT constructs lacking the enhancers were inactive in the transgenic mice. In one case, a truncated δ2-crystallin promoter (-308/+24) in combination with the enhancer was also active in the Purkinje cells of the cerebellum of the transgenic mice, which could prove useful in future experiments. Finally, retinoic acid receptors (RARβ) activated the δ1-crystallin, but not the δ2-crystallin enhancer in the recombinant plasmids in cotransfected embryonic chicken lens epithelial cells treated with retinoic acid. This activation did not occur when using the core enhancer (fragment B4)lacking surrounding flanking sequences (fragment B3 and B5) of the enhancer. Together these experiments show that the chicken δ-crystallin enhancers show lens-preference in transgenic mice despite the absence of δ-crystallin in this species and add retinoic acid nuclear receptors to the growing list of transcription factors (including Pax-6, Sox-2, and δEF3) that directly or indirectly contribute to the high expression of the δ1-crystallin gene in the lens.

Original languageEnglish (US)
Pages (from-to)258-266
Number of pages9
JournalDevelopmental Genetics
Volume20
Issue number3
DOIs
StatePublished - 1997
Externally publishedYes

Fingerprint

Crystallins
Tretinoin
Transgenic Mice
Lenses
Chickens
Genes
Retinoic Acid Receptors
Chloramphenicol O-Acetyltransferase
Paired Box Transcription Factors
Purkinje Cells
Cytoplasmic and Nuclear Receptors
Reporter Genes
Introns
Cerebellum
Transfection
Plasmids
Epithelial Cells

Keywords

  • δ-Crystallin
  • Enhancers
  • Gene expression
  • Lens
  • Retinoic acid
  • Transgenic mice

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology
  • Genetics

Cite this

Lens-preferred activity of chicken δ1- and δ2-crystallin enhancers in transgenic mice and evidence for retinoic acid-responsive regulation of the δ1-crystallin gene. / Li, Xuan; Cvekl, Ales; Bassnett, Steven; Piatigorsky, Joram.

In: Developmental Genetics, Vol. 20, No. 3, 1997, p. 258-266.

Research output: Contribution to journalArticle

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abstract = "There are two tandemly linked δ-crystallin genes (5' δ-δ2 3') in the chicken, with the δ1-crystallin gene being expressed much more highly (50- 100-fold) in the embryonic lens than the δ2-crystallin gene. Previous transfection experiments have shown that a lens preferred enhancer exists in the third intron of each chicken δ-crystallin gene. In the present investigation we have used transgenic mice to establish that both the chicken δ1- and δ2-crystallin enhancers are preferentially active in the mouse lens in combination with their homologous promoter and the chloramphenicol acetyltransferase (CAT) reporter gene. The promoter/CAT constructs lacking the enhancers were inactive in the transgenic mice. In one case, a truncated δ2-crystallin promoter (-308/+24) in combination with the enhancer was also active in the Purkinje cells of the cerebellum of the transgenic mice, which could prove useful in future experiments. Finally, retinoic acid receptors (RARβ) activated the δ1-crystallin, but not the δ2-crystallin enhancer in the recombinant plasmids in cotransfected embryonic chicken lens epithelial cells treated with retinoic acid. This activation did not occur when using the core enhancer (fragment B4)lacking surrounding flanking sequences (fragment B3 and B5) of the enhancer. Together these experiments show that the chicken δ-crystallin enhancers show lens-preference in transgenic mice despite the absence of δ-crystallin in this species and add retinoic acid nuclear receptors to the growing list of transcription factors (including Pax-6, Sox-2, and δEF3) that directly or indirectly contribute to the high expression of the δ1-crystallin gene in the lens.",
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