Regeneration

March 13, 2015

János Szabad
Department of Biology
Faculty of Medicine
University of Szeged
Somogyi str. 4
H-6720 Szeged, Hungary
E-mail: szabad.janos@med.u-szeged.hu
 Features of mitotic recombination
- Way to genetically label cell(s); twin and single spots
- Mitotic recombination, a source of tumorigenesis
- Tool to analyze organ development
- Autonomy, non-autonomy, perdurnce
- Minute technique and the compartments
- Analysis of engrailed (en) gene function
- Labeling germ-line stem cells
- Tool to detect and measure DNA breaks
- Tool to detect and measure cell death
 Mitotic recombination

Cells in a Drosophila wing

primordium

Mitosis
Replication X-rays

Centromere mwh

mwh mwh

+ +

Mother cell Mitotic +

recombination Daughter cells

Clone size Clone size, cell death and regeneration

- 40-60% of the wing imaginal disc cells are lost following

irradiation by 1000 R.
- The lost cells are replaced through intercalary regeneration.
Haynie and Bryant, Roux’s Arch. Dev. Biol. 183, 85, 1977.

- While soft X-rays kill many cells and induce few mitotic
recombinations, hard X-ray kill few cells and induce many
mitotic recombination.
Kumar, Thesis, University of Szeged, 1978.
← If there was no cell death
2,000 4,000
Dose (Rad)
 Intercalary regeneration
If there is no
cell death

A genetically Cell death followed by

labeled cell
intercalary regeneration

Dying cell
 Drosophila imaginal discs
Larva Imaginal discs Adult
Mouth
parts

Antenna
Eye

Leg

Wing

Haltere

Terminalia
Drosophila wing imaginal discs and its derivatives:
wing and notum
When transplanted into matured larvae, the discs fragments
go through metamorphosis, can be recovered and analyzed.






Metamorphosis










Analyze and
construct
fate map

Peter J. Bryant, J Exp Zool 193, 49, 1975.

Drosophila wing disc fate map

Bryant, J Exp Zool 193, 49, 1975.

 Recover Recover
implant implant

Analyze
Transplant

Metamorphosis
Transplant into female Duplicated
 abdomen to culture

Cut a
wing disc
into two

Recover Recover
implant implant

Transplant Analyze

Regenerated
 A The downhill principle

Developmental
B

capacity
C
D High point
E
Position
A
B Cut into two

C C
D
A E
B
C C
D D D
E E E
Regeneration Duplication
Bryant, J Exp Zool 193, 49, 1975.
 Disc fragments and intercalary regeneration

 Intercalary
regeneration

Middle fragment
discarded

 Intercalary
regeneration

X-irradiate
to kill cells
The polar coordinate model



Polar coordinate and
positional information

Regeneration follows two rules:

1. Shortest-route intercalation
When cells with noncontiguous values are juxtaposed growth occurs until
cells with the intervening positional values are intercalated.
2. Complete circle rule for distal regeneration
Cells at any radial level can give rise to cells with distal positional values as long
as a complete circular sequence of values is present at the cut edge.
Bryant, Bryant and French, Sci Am 237, 66, 1977.
Bryant, Bryant and French, Sci Am 237, 66, 1977.
 Regeneration in hydras

Regeneration Growth
Amputation

Hydra
Regeneration in the planaria flatworm
 Intercalary
regeneration

Initial condition

Right after grafting

Outcome
Young limb bud
Mesoderm
Apical Ectodermal Ridge controls
AER growth in the proximal-distal
Ectoderm
direction
Old limb bud

Old limb bud

Young limb bud

 A newt can regenerate an
entire limb in 7-10 weeks

One week

3-6 weeks

6-9 weeks
A vestige of limb regeneration in humans
Young children’s cleanly
sheared off beyond this level
will regenerate perfectly

Before After Before After

http://en.mebo.com/UpLoadFiles/Article/2009-5/2009051213245533739.jpg
 Mouth
parts Transdetermination
Antenna
Eye

Leg

Wing

Haltere

Terminalia

McClure and Schubiger, Int J Biochem Cell Biol 39, 1105, 2007.
Transdetermination

McClure and Schubiger, Int J Biochem Cell Biol 39, 1105, 2007.