title: Mouse conjunctival forniceal gene expression during postnatal development and its regulation by Kruppel-like factor 4.
abstract: OBJECTIVE
  To identify the changes in postnatal mouse conjunctival forniceal gene expression and their regulation by Klf4 during the eye-opening stage when the goblet cells first appear.
  
  
  METHODS
  Laser microdissection (LMD) was used to collect conjunctival forniceal cells from postnatal (PN) day 9, PN14 and PN20 wild-type (WT), and PN14 Klf4-conditional null (Klf4CN) mice, in which goblet cells are absent, developing, present, and missing, respectively. Microarrays were used to compare gene expression among these groups. Expression of selected genes was validated by quantitative RT-PCR, and spatiotemporal expression was assessed by in situ hybridization.
  
  
  RESULTS
  This study identified 668, 251, 1160, and 139 transcripts that were increased and 492, 377, 1419, and 57 transcripts that were decreased between PN9 and PN14, PN14 and PN20, PN9 and PN20, and PN14 WT and Klf4CN conjunctiva, respectively. Transcripts encoding transcription factors Spdef, FoxA1, and FoxA3 that regulate goblet cell development in other mucosal epithelia, and epithelium-specific Ets (ESE) transcription factor family members were increased during conjunctival development. Components of pathways related to the mesenchymal-epithelial transition, glycoprotein biosynthesis, mucosal immunity, signaling, and endocytic and neural regulation were increased during conjunctival development. Conjunctival Klf4 target genes differed significantly from the previously identified corneal Klf4 target genes, implying tissue-dependent regulatory targets for Klf4.
  
  
  CONCLUSIONS
  The changes in gene expression accompanying mouse conjunctival development were identified, and the role of Klf4 in this process was determined. This study provides new probes for examining conjunctival development and function and reveals that the gene regulatory network necessary for goblet cell development is conserved across different mucosal epithelia.
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Eye and Ear Institute, 203 Lothrop Street, Pittsburgh PA 15213, USA.
authors.forename: Divya
authors.initials: D
authors.lastname: Gupta
authors.forename: Stephen A K
authors.initials: SA
authors.lastname: Harvey
authors.forename: Naftali
authors.initials: N
authors.lastname: Kaminski
authors.forename: Shivalingappa K
authors.initials: SK
authors.lastname: Swamynathan
mesh_terms: D000818:Animals; D003228:Conjunctiva / Q000254:growth & development* / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005838:Genotype; D020397:Goblet Cells / Q000166:cytology; D017403:In Situ Hybridization; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008297:Male; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics*
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D014157:Transcription Factors