The Corneal Epithelial Barrier and Its Developmental Role in Isolating Corneal Epithelial and Conjunctival Cells From One Another.

Purpose: During development, the corneal epithelium (CE) and the conjunctiva are derived from the surface ectoderm. Here we have examined how, during development, the cells of these two issues become isolated from each other.

Methods: Epithelia from the anterior eyes of chicken embryos were labeled with the fluorescent, lipophilic dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI).

Nuclear ferritin mediated regulation of JNK signaling in corneal epithelial cells.

Corneal epithelial (CE) cells are exposed to environmental insults (e.g., UV-irradiation), yet they suffer little damage.

Developmental regulation of trigeminal TRPA1 by the cornea.

Purpose: The cornea is densely innervated with nociceptive nerves that detect deleterious stimuli at the ocular surface and transduce these stimuli as sensations of pain. Thus, nociception is a major factor involved in preventing damage to corneal tissues. One class of molecules that is thought to be involved in detecting such stimuli is the transient receptor potential (TRP) family of ion channels.

Developmental guidance of embryonic corneal innervation: roles of Semaphorin3A and Slit2.

The cornea is one of the most densely innervated structures of the body. In the developing chicken embryo, nerves from the ophthalmic trigeminal ganglion (OTG) innervate the cornea in a series of spatially and temporally regulated events. However, little is known concerning the signals that regulate these events.

Developmental corneal innervation: interactions between nerves and specialized apical corneal epithelial cells.

Purpose: The corneal epithelium is one of the most highly innervated structures in the body, and proper innervation is necessary for corneal maintenance and sensation. However, little is known about how these nerves function and how innervation occurs developmentally. The authors have examined certain aspects of corneal innervation in the developing chicken embryo.

Developmental regulation of the nuclear ferritoid-ferritin complex of avian corneal epithelial cells: roles of systemic factors and thyroxine.

Previously we observed that avian corneal epithelial cells protect their DNA from oxidative damage by having the iron-sequestering molecule ferritin - normally cytoplasmic - in a nuclear location. This localization involves a developmentally-regulated ferritin-like protein - ferritoid - that initially serves as the nuclear transporter, and then as a component of a ferritoid-ferritin complex that is half the size of a typical ferritin and binds to DNA. We also observed that developmentally, the synthesis of ferritin and ferritoid are regulated coordinately - with ferritin being predominantly translational and ferritoid transcriptional.

Phosphorylation regulates the ferritoid-ferritin interaction and nuclear transport.

Ferritin is an iron-sequestering protein that is generally cytoplasmic; however, our previous studies have shown that in avian corneal epithelial (CE) cells ferritin is nuclear. We have also observed that this nuclear localization involves a tissue-specific nuclear transporter that we have termed ferritoid, and that nuclear ferritin protects DNA from oxidative damage. Recently we have determined that ferritoid functions not only as a nuclear transporter, but also, within the nucleus, it remains associated with ferritin as a heteropolymeric complex.

Nuclear ferritin: a ferritoid-ferritin complex in corneal epithelial cells.

Purpose: Ferritin is an iron storage protein that is generally cytoplasmic. However, in embryonic avian corneal epithelial (CE) cells, the authors previously observed that the ferritin was predominantly nuclear. They also obtained evidence that this ferritin protects DNA from oxidative damage by UV light and hydrogen peroxide and that the nuclear localization involves a tissue-specific nuclear transporter, termed ferritoid.

Corneal epithelial nuclear ferritin: developmental regulation of ferritin and its nuclear transporter ferritoid.

The corneal epithelium is exposed to reactive oxygen species that are potentially deleterious to nuclear DNA. However, our previous studies show that corneal epithelial cells have a novel, developmentally regulated mechanism for protection from such damage that involves having the iron-sequestering molecule, ferritin, in the nucleus. Nuclear localization of ferritin is achieved through the action of a tissue-specific nuclear transporter, ferritoid, which is itself a ferritin family member.

Identification of unique molecular subdomains in the perichondrium and periosteum and their role in regulating gene expression in the underlying chondrocytes.

Developing cartilaginous and ossified skeletal anlagen is encapsulated within a membranous sheath of flattened, elongated cells called, respectively, the perichondrium and the periosteum. These periskeletal tissues are organized in distinct morphological layers that have been proposed to support distinct functions. Classical experiments, particularly those using an in vitro organ culture system, demonstrated that these tissues play important roles in regulating the differentiation of the subjacent skeletal elements.

Nuclear ferritin-mediated protection of corneal epithelial cells from oxidative damage to DNA.

We previously obtained evidence that ferritin is a nuclear protein in embryonic avian corneal epithelial (CE) cells, and that the ferritin in this site protects DNA from UV-induced damage. UV irradiation is known to produce reactive oxygen species (ROS) and ferritin is known to ameliorate further oxidative damage by sequestering free iron, thus decreasing the formation of hydroxyl radicals through the Fenton reaction. Here we present evidence that nuclear ferritin can similarly prevent damage by the ROS, H2O2.

Perichondrial-mediated TGF-beta regulation of cartilage growth in avian long bone development.

We previously observed using cultured tibiotarsal long-bone rudiments from which the perichondrium (PC) and periosteum (PO) was removed that the PC regulates cartilage growth by the secretion of soluble negative regulatory factors. This regulation is "precise" in that it compensates exactly for removal of the endogenous PC and is mediated through at least three independent mechanisms, one of which involves a response to TGF-beta. PC cell cultures treated with 2 ng/ml TGF-beta1 produced a conditioned medium which when added to PC/PO-free organ cultures effected precise regulation of cartilage growth.

Identification and characterization of a previously undetected region between the perichondrium and periosteum of the developing avian limb.

In developing long bones, the growing cartilage and bone are surrounded by the fibrous perichondrium (PC) and periosteum (PO), respectively, which provide cells for the appositional growth (i.e., growth in diameter) of these tissues.

SP3/SP1 transcription activity regulates specific expression of collagen type X in hypertrophic chondrocytes.

Previously, we have shown that two non-canonical specificity protein (SP)-binding sites within the proximal promoter (nucleotide (nt) -139 to +5) of the chicken Col10a1 gene are involved in conferring tissue-specific expression of type X collagen to hypertrophic chondrocytes. In the present study, we examined the role of SP3/SP1 transcription factors in the regulation of the Col10a1 promoter. The SP3/SP1 ratio is higher in hypertrophic versus non-hypertrophic chondrocytes, due to the significant decrease in SP1 in hypertrophic cells detected by real-time PCR and Western blot analyses.

Nuclear ferritin in corneal epithelial cells: tissue-specific nuclear transport and protection from UV-damage.

We have identified the heavy chain of ferritin as a developmentally regulated nuclear protein of embryonic chicken corneal epithelial cells. The nuclear ferritin is assembled into a supramolecular form that is indistinguishable from the cytoplasmic form of ferritin found in other cell types. Thus it most likely has iron-sequestering capabilities.

Cellular invasion of the chicken corneal stroma during development: regulation by multiple matrix metalloproteases and the lens.

Avian corneal development requires cellular invasion into the acellular matrix of the primary stroma. Previous results show that this invasion is preceded by the removal of the fibril-associated type IX collagen, which possibly stabilizes matrices through interfibrillar cross-bridges secured by covalent crosslinks. In the present study, we provide evidence for the expression of three matrix metalloproteinases (MMPs) in early corneas, two of which act cooperatively to selectively remove type IX collagen in situ.

Chondrocyte-derived transglutaminase promotes maturation of preosteoblasts in periosteal bone.

During endochondral development, elongation of the bone collar occurs coordinately with growth of the underlying cartilaginous growth plate. Transglutaminases (TGases) are upregulated in hypertrophic chondrocytes, and correlative evidence suggests a relationship between these enzymes and mineralization. To examine whether TGases are involved in regulating mineralization/osteogenesis during bone development, we devised a coculture system in which one cellular component (characterized as preosteoblastic) is derived from the nonmineralized region of the bone, and the other cellular component is hypertrophic chondrocytes.

Ferritoid, a tissue-specific nuclear transport protein for ferritin in corneal epithelial cells.

Previously we reported that ferritin in corneal epithelial (CE) cells is a nuclear protein that protects DNA from UV damage. Since ferritin is normally cytoplasmic, in CE cells, a mechanism must exist that effects its nuclear localization. We have now determined that this involves a nuclear transport molecule we have termed ferritoid.

Positive regulation of endochondral cartilage growth by perichondrial and periosteal calcitonin.

Our previous studies showed that during the embryonic development of avian long bones, growth of the cartilaginous component is regulated by multiple factors secreted by the surrounding perichondrium (PC) and periosteum (PO). The activities of these factors--which include both positive and negative regulators--can be detected in conditioned media from PC and PO cell cultures. In the present study, we have obtained evidence suggesting that a positive regulator is the peptide hormone calcitonin (CT).

Multiple mechanisms of perichondrial regulation of cartilage growth.

We previously observed that the perichondrium (PC) and the periosteum (PO) negatively regulate endochondral cartilage growth through secreted factors. Conditioned medium from cultures of PC and PO cells when mixed (PC/PO-conditioned medium) and tested on organ cultures of embryonic chicken tibiotarsi from which the PC and PO have been removed (PC/PO-free cultures) effect negative regulation of growth. Of potential importance, this regulation compensates precisely for removal of the PC and PO, thus mimicking the regulation effected by these tissues in vivo.