TISSUE ENGINERRING DIGITAL ASSIGNMENT 1

Name – Sona Elizabeth Binoy

Reg.no – 20BBT0300
Guided by –Dr. MANJUBALA I

MORPHOGENESIS OF TISSUE

INTRODUCTION
Morphogenesis is the process by which the individual cells within the
developing embryo move around and organize themselves to form the
structures, organs and systems that make up the adult organism.
Morphogenesis is directed by two types of factors - chemical factors
(molecules) and mechanical forces.
One particularly important group of molecules involved in morphogenesis
are the morphogens. Morphogens are signal molecules that can affect the
internal processes within the cell as well as cell behaviour and cell
movement.

Cells tend to respond to morphogens based on their local concentration.

Morphogens generally bind to cell receptors on the membrane, thereby
initiating some type of intracellular response.
Typically, a molecule called the transcription factor goes into the nucleus
and attaches to the DNA. This in turn either turns on or turns off the
expression of some sort of gene.
The protein produced from expressing the gene can then change the
composition of the extracellular membrane, change the cell-to-cell
adhesion properties and contract the cell.
Changing the cell adhesion properties can detach the cell form another
cell and changing the extracellular properties can allow the cell to move
to another location within that developing tissue. In this matter, the embryo
can gradually form the tissues and organs of that organism.

MORPHOGENESIS OF EYE
Eye formation in the human embryo begins at approximately three weeks
into embryonic development and continues through the tenth week. The
formation of the eye involves cells from both the ectodermal and
mesodermal tissues.
In particular, the neuroepithelium, surface ectoderm, and extracellular
mesenchyme, which includes both neural crest and mesoderm, are the
sources of the eye.
Neuroepithelium forms the retina, ciliary body, iris, and optic nerves.
Surface ectoderm forms the lens, corneal epithelium and eyelid.
The extracellular mesenchyme forms the sclera, the corneal endothelium
and stroma, blood vessels, muscles, and vitreous.
The eye begins to develop as a pair of optic vesicles on each side of the
forebrain at the end of the 4th week of pregnancy.

Optic vesicles are outgrowing’s of the brain which make contact with the
surface ectoderm and this contact induces changes necessary for further
development of the eye.
Through a groove at the bottom of the optic vesicle known as choroid
fissure the blood vessels enter the eye.

Three embryonic tissue sources: the neural ectoderm, the surface

ectoderm, and the periocular mesenchyme contribute to the formation of
the mammalian eye.

REGULATION OF EYE DEVELOPMENT

Eye development is initiated by the master control gene PAX6, a
homeobox gene with known homologues in humans (aniridia), mice (small
eye), and Drosophila (eyeless).

The PAX6 gene locus is a transcription factor for the various genes and
growth factors involved in eye formation.

Eye morphogenesis begins with the evagination, or outgrowth, of the optic

grooves or sulci. These two grooves in the neural folds transform into optic
vesicles with the closure of the neural tube.

The optic vesicles then develop into the optic cup with the inner layer
forming the retina and the outer portion forming the retinal pigment
epithelium. The middle portion of the optic cup develops into the ciliary
body and iris.

During the invagination of the optic cup, the ectoderm begins to thicken
and form the lens placode, which eventually separates from the ectoderm
to form the lens vesicle at the open end of the optic cup.
Figure: Early ocular morphogenesis
(A) Developmental pathways such as Wnt, BMP, and fibroblast growth factor (FGF)
drive upregulation of eye-field transcription factors in the anterior neural plate, creating
the specified region known as the "eye-field."
(B) Deepening of optic sulci and evagination of optic vesicle around 22 days post-
conception. The newly formed optic vesicle ubiquitously expresses all eye-field
transcription factors.
(C) The action of signalling pathways determines presumptive regions in the optic
vesicle characterized by unique gene expression patterns in the third and 4th weeks
of gestation.
(D) Interactions between the optic vesicle, surface ectoderm, and extraocular
mesenchyme cause the invagination of the optic cup at approximately 5 weeks post-
conception. MITF and VSX2 interactions create boundaries between retinal pigment
epithelium (RPE) and neuroretina (NR) in the developing optic cups. The lens pit
begins to form from the surface ectoderm.
(E) In the 5th week of gestation following optic cup formation, Wnt and FGF pathways
drive RPE/NR differentiation and clear definition of these regions through ciliary
margin formation. The lens vesicle forms as the lens pit detach from the surface
ectoderm.
(F) By the 7th week of gestation, lens fibres extend to form the lens from the hollow
lens vesicle. The cornea forms from the overlying surface ectoderm. NR and RPE are
clearly defined and separated by the ciliary margins, while the optic nerve forms from
the convergence of the optic stalk.

OPTIC CUP MORPHOGENESIS

The distal portion of the optic vesicle makes contact with the overlying
surface ectoderm, resulting in the specification of the lens ectoderm (pre-
placodal stage). This interaction leads to invagination of the lens placode
and distal optic vesicle resulting in formation of a bilayer optic cup.
The neural retina and RPE develop from the inner and outer layer of the
optic cup, respectively. The lens vesicle eventually separates from the
surface ectoderm and differentiates into the mature lens.

Figure: Invagination of the lens placode requires correct specification of

the lens ectoderm that is dependent on Six3-mediated maintenance and
activation of Pax6 and Sox2, respectively. FGF and BMP signalling may
be also required for lens induction.
DEVELOPMENT OF RETINA
The two layers of the optic cup will further differentiate into the retina of
the mature eye. The two layers are unequal in size - the outer one is
thinner than the inner one.
The optic cup can be divided into two portions, the anterior 1/5 (rim) and
the posterior 4/5. The rim area will ultimately form the iris and ciliary body,
and the posterior 4/5 will form the retina.
The outer layer of the posterior 4/5 will become the pigment layer of the
retina, and the inner one will become the neural retina. These two layers
are separated by the intraretinal space.

The development of the retina’s pigment layer is very straightforward, with

the appearance of melanin granules in the cells of this layer at around 4
1/2 weeks. Slightly later, at about 6 weeks, the cells in the posterior aspect
of the inner layer of the optic cup begin a more complicated process.
The cells immediately adjacent to the intraretinal space begin to
differentiate into the photoreceptors (rods and cones). The next layer of
cells will become the Muller supporting cells and the bipolar neurons, and
the innermost superficial layer will develop into the axons of the ganglion
cells, the ones that will make up the optic nerve.
This means light actually passes through the neuronal layers before
reaching the rods and cones. The ganglion cell fibres gradually fill in the
lumen of the optic stalk as it becomes the optic nerve.
By eight months, all the layers of the retina are recognizable. But
maturation of the photoreceptors continues after birth, which in part
explains why a baby’s visual acuity improves as it grows.

LENS DEVELOPMENT
Lens development is closely related to optic vesicle development. The
interaction between the growing vesicle and the ectoderm causes the
ectoderm to thicken at that point. This thickened portion of the ectoderm
is called the lens placode.
Next, the placode invaginates and forms a pouch referred to as the lens
pit. Eventually, the pit becomes completely enclosed. This enclosed
structure is the lens vesicle.
Then at about the same time as the pigmented layer of the retina is
developing, the cells of the posterior part of the lens vesicle transform into
elongated, slender primary lens fibers. These new cells fill in the
previously hollow structure.
About four weeks later, more lens fibers develop, this time from the
anterior wall of the lens vesicle (secondary lens fibers).

DEVELOPMENT OF THE CHOROID, SCLERA & CORNEA

During the sixth and seventh weeks the mesenchyme that surrounds the
external surface of the optic cup condenses into two layers, an inner,
pigmented, vascular layer known as the choroid and an outer, fibrous layer
called the sclera.
The mesenchyme that is anterior to the developing lens splits into two
layers that surround the newly formed anterior chamber of the eye.
The inner layer is continuous with the choroid and is called the
iridopupillary membrane and the outer layer is continuous with the sclera.
The outer layer will form the substantia propria, or stroma of the cornea.
The cornea has three layers, epithelium, stroma, and endothelium. The
external corneal epithelium develops from surface ectoderm and the
endothelium forms from neural crest cells that migrate from the rim of the
optic cup. The stroma is derived from the surrounding mesenchyme. The
iridopupillary membrane eventually disappears completely, which allows
communication between the anterior and posterior eye chambers.
DEVELOPMENT OF THE IRIS AND CILIARY BODY
The anterior rim of the optic cup gives rise to the epithelium of the iris and
the ciliary body. Remember that the inner layer of the posterior 4/5 of the
optic cup forms the neural retina of the eye. The anterior part of this inner
layer forms the non-pigmented layer of the iris and the ciliary process
epithelium. The outer layer of the optic cup in this region contributes the
pigmented epithelial layer. A few folds form in the anterior aspect of the
optic cup and this forms the ciliary processes.

DERIVATIVES OF VARIOUS TISSUES

Neuroectoderm:
1. Pigmented epithelia of retina (1 layer), ciliary body (1 layer), and iris (2
layers).
2. Sensory retina, and innermost (nonpigmented) layer of ciliary body
3. Optic nerve
4. Iris sphincter and dilator muscles
5. Vitreous (part)
Surface ectoderm:
1. Lens
2. Corneal epithelium
3. Conjunctiva and caruncle
4. Eyelid skin
5. Lacrimal apparatus (glands and drainage system)
Head mesenchyme (neural crest and/or mesoderm):
1. Blood vessels
2. Corneal stroma and endothelium
3. Stroma of choroid, ciliary body, and iris
4. Ciliary muscle
5. Sclera
6. Optic nerve sheath (meninges)
7. Extraocular muscles and fasciae
8. Remainder of the eyelids (orbicularis oculi muscle, tarsus, orbital
septum, etc)
9. Vitreous (part)

CONCLUSION
The mature eye provides a valuable model for cellular replacement and
regenerative therapies. It is surgically accessible and nonessential for life,
and failed tissue grafts can be ablated or removed with relative ease.
Moreover, advanced imaging technologies for evaluating the structure
and function of the adult eye, and the wealth of basic knowledge of eye
field and retinal development, together have enhanced the field of
regenerative medicine for the treatment of currently incurable eye
disease.
The eye has served as an invaluable model for understanding the
mechanisms that coordinate human embryogenesis. Developmental
genes can be identified readily through ocular phenotypes, because the
eye is not essential for the survival of the organism. Continued
characterization of previously unidentified eye development genes and
persistent dialogue between basic scientists and clinicians will serve as a
paradigm for understanding fundamental processes in human
development and treating degenerative disease.
REFERENCES
1. Ashery-Padan R, Marquardt T, Zhou X, Gruss P. 2000. Pax6 activity
in the lens primordium is required for lens formation and for correct
placement of a single retina in the eye. Genes Dev 14: 2701–2711.

2. Beebe DC. 1986. Development of the ciliary body: A brief

review. Trans Ophthalmol Soc UK 105: 123–130.

3. Bringmann A, Pannicke T, Grosche J, Francke M, Wiedemann P, Skat

chkov SN, Osborne NN, Reichenbach A. 2006. Muller cells in the
healthy and diseased retina. Prog Retin Eye Res 25: 397–424.

4. Hagglund AC, Dahl L, Carlsson L. 2011. Lhx2 is required for

patterning and expansion of a distinct progenitor cell population
committed to eye development. PLoS ONE 6: e23387.

5. Hendrix RW, Zwaan J. 1975. The matrix of the optic vesicle-

presumptive lens interface during induction of the lens in the chicken
embryo. J Embryol Exp Morphol 33: 1023–1049.

6. Hever AM, Williamson KA, van Heyningen V. 2006. Developmental

malformations of the eye: The role of PAX6, SOX2 and OTX2. Clin
Genet 69: 459–470.

7. Saint-Geniez M, D’Amore PA. 2004. Development and pathology of

the hyaloid, choroidal and retinal vasculature. Int J Dev Biol 48: 1045–
1058.

8. Zaghloul NA, Yan B, Moody SA. 2005. Step-wise specification of

retinal stem cells during normal embryogenesis. Biol Cell 97: 321–337.