Role In Ocular Development Research Articles

Heat shock protein 108 mRNA expression during chicken retina development

In a developmental study on the expression of heat shock protein 108 (HSP108) mRNA in the chicken retina, we found different spatial and temporal expressions of HSP108 mRNA in each retinal layer. While intense HSP108 signals were found in the retina neuroblast layer at embryonic day 5 (E5), the ganglion cell population (GC), inner nuclear layer (IN) and pigment epithelium (PE) showed HSP108 expression at E9. At E14, HSP108 signals were reduced versus the previous stages even though signals were still detected in the GC, the IN, the outer nuclear layer and the PE. HSP108 signals were still detectable at the E21 stage, although each retinal layer showed a much differentiated morphology and diminished signal intensity. These results suggest that HSP108 expression might be developmentally regulated throughout eye organogenesis and that it plays a role in ocular development.

Identification of a novel GH isoform: a possible link between GH and melanocortin systems in the developing chicken eye.

GH is considered to play a role in the pathogenesis of diabetic retinopathy, causing neovascularization in the retina. The present study was conducted to assess the possibility that GH may play a role in ocular development by determining whether GH is expressed in the eye of the chicken during development. In the 17-d-old embryo, immunocytochemistry detected immunoreactive GH in retinal pigment epithelial (RPE) cells. Characterization of GH mRNA expressed in the eye by RT-PCR and rapid amplification of cDNA 5'-ends revealed it to be a novel GH mRNA transcribed from the middle of the intron 3 of the chicken GH (cGH) gene. The deduced protein, designated small GH isoform (s-cGH), was a cytosolic protein of 16.5 kDa with 140 amino acid (aa) residues, lacking the signal peptide and the N-terminal 71 aa residues of 22-kDa cGH, replacing them with 20 aberrant aa residues, and identical to 22-kDa cGH for the C-terminal 120-aa residue portion. Western blotting determined the molecular size of immunoreactive GH in RPE cells to be 80-84 kDa, similar to the computed molecular mass of s-cGH/GH receptor complex. Furthermore, RT-PCR demonstrated that GH receptor mRNA, but not s-cGH mRNA, was expressed in RPE cells. These results suggest that RPE cell is one of the target cells of s-cGH in the eye. During embryonic development, the immunoreactivity for s-cGH in RPE cells was initially observed on embryonic d 10, and the staining intensity increased and peaked on embryonic d 17. By hatching, s-cGH immunoreactivity in RPE cells was gradually decreased, and it was not detectable after hatching. This ontogenetic staining pattern correlates well with the pattern of the production of alpha MSH in RPE cells. The cell type expressing s-cGH remains to be identified; however, our findings imply a possible involvement of GH in the regulation of ocular development by acting on the intraocular melanocortin system in the chicken.

Pax6 activity in the lens primordium is required for lens formation and for correct placement of a single retina in the eye

The Pax6 transcription factor plays a key role in ocular development of vertebrates and invertebrates. Homozygosity of the Pax6 null mutation in human and mice results in arrest of optic vesicle development and failure to initiate lens formation. This phenotype obscures the understanding of autonomous function of Pax6 in these tissue components and during later developmental stages. We employed the Cre/loxP approach to inactivate Pax6 specifically in the eye surface ectoderm concomitantly with lens induction. Although lens induction occurred in the mutant, as indicated by Sox2 up-regulation in the surface ectoderm, further development of the lens was arrested. Hence, Pax6 activity was found to be essential in the specified ectoderm for lens placode formation. Furthermore, this mutant model allowed us for the first time to address in vivo the development of a completely normal retina in the absence of early lens structures. Remarkably, several independent, fully differentiated neuroretinas developed in a single optic vesicle in the absence of a lens, demonstrating that the developing lens is not necessary to instruct the differentiation of the neuroretina but is, rather, required for the correct placement of a single retina in the eye.

Mutations of the Forkhead/Winged-Helix Gene, FKHL7, in Patients with Axenfeld-Rieger Anomaly

Genetic linkage, genome mismatch scanning, and analysis of patients with alterations of chromosome 6 have indicated that a major locus for development of the anterior segment of the eye, IRID1, is located at 6p25. Abnormalities of this locus lead to glaucoma. FKHL7 (also called "FREAC3"), a member of the forkhead/winged-helix transcription-factor family, has also been mapped to 6p25. DNA sequencing of FKHL7 in five IRID1 families and 16 sporadic patients with anterior-segment defects revealed three mutations: a 10-bp deletion predicted to cause a frameshift and premature protein truncation prior to the FKHL7 forkhead DNA-binding domain, as well as two missense mutations of conserved amino acids within the FKHL7 forkhead domain. Mf1, the murine homologue of FKHL7, is expressed in the developing brain, skeletal system, and eye, consistent with FKHL7 having a role in ocular development. However, mutational screening and genetic-linkage analyses excluded FKHL7 from underlying the anterior-segment disorders in two IRID1 families with linkage to 6p25. Our findings demonstrate that, although mutations of FKHL7 result in anterior-segment defects and glaucoma in some patients, it is probable that at least one more locus involved in the regulation of eye development is also located at 6p25.

Messenger RNA expression of heat shock proteins (HSPs) during ocular development

The heat shock proteins (HSPs) are believed to act as molecular chaperones which appear to play some roles in regulation of normal protein folding and also in preventing damage to protein structures under various conditions of environmental stress. We examined the expression of the major HSP families, HSP60, 70 and 90 families and small HSP32, at the mRNA level in ocular development. Expression of HSP32, HSP60, HSP70, HSP84, HSP86 and heat shock cognate protein (HSC)70 mRNAs was examined by in situ hybridization. HSC70, HSP84, HSP86 and HSP60 mRNAs were expressed strongly in all ocular tissues during early stages 3 to 5, corresponding to embryonic day (E)11.5 to E14.5. At stages 6 to 7 (E15.5 to E18.5), the expression of these four mRNA species was decreased markedly in most ocular tissue, while in the retina strong HSC70 and HSP86 mRNA expression was still detected. HSP32 and HSP70 mRNAs were not detected at any stage. These results suggest that the expression of HSPs is developmentally regulated through ocular organogenesis, and the proteins may play some important roles in ocular development.