T3 Limb Flashcards

1
Q

In early experiments of salamander limb development, what is the outcome if you remove half the limb disc?

A

Remaining half will form a complete limb

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2
Q

What is the limb disc? salamander

A

it is the thickened area on the side of the salamander that will give rise to the limb.

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3
Q

What is the actual primodium for the limb? salamander

A

the limb disc

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4
Q

The peripheral region of salamander gives rise to what?

A

it is the region around the disc is the morphogenetic field and can give rise tot he limb if the disc is removed.

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5
Q

What is the outcome of the limb if the limb field is removed? salamanders

A

no limb will form

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6
Q

In early experiments of salamander limb development, what is the outcome if you vertically divide the disc into two halves and physically separate the two halves?

A

each half will form a normal limb

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7
Q

In early experiments of salamander limb development, what is the outcome if you fuse two discs together?

A

a single, normal limb will form

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8
Q

What initiate limb development?

A

paraxial mesoderm signals via retinoic acid based on hox code.

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9
Q

What are the initial initiators of limb development?

A

Tbx5 and Tbx4

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10
Q

What does the expression of Tbx5 initiate?

A

the future forelimb

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11
Q

What does the expression of Tbx4 and Pitx-1 initiate?

A

the future hindlimb

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12
Q

The expression of what two transcription factors fixes the anterior-posterior axis?

A

Gli-3 and Hand-2

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13
Q

What is associated with fixing the anterior portion of the anterior-posterior axis?

A

Gli-3

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14
Q

What is associated with fixing the posterior portion of the anterior-posterior axis?

A

Hand-2

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15
Q

What are the three axes that define the vertebrate limb?

A

dorsoventral, anterior-posterior, and proximodistal

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16
Q

Early research on establishment of the three axes did what?

A
  1. transplantation experiments allowed workers to reverse one or more axes.
  2. transplant a limb disc to the opposite side
  3. rotate a limb disc 180 degrees
  4. Slack in 1976 transplanted a strip of flank tissue from just posterior to the limb disc to an ectopic site just anterior to the limb disc. This causes the limb to consist of two posterior halves arranged in a mirror image fashion
  5. Riddle (1993) suggested that the morphogen expression by the posterior organization region might be the RNA product of the shh gene. Treatments of the anterior part with retinoic acid caused cells in this region to behave like cells in the posterior organization region. Retinoic acid was not the morphogen but was/is capable of activating shh, which may be the actual morphogen.
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17
Q

What is the function of signaling molecule Hoxb8?

A

determines the position of the zone of polarizing activity (ZPA) via its highest concentration, and its expression is induced by retinoic acid.

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18
Q

What is the function of signaling molecule Shh?

A
  • it is a signal produced by ZPA
  • maintains the structure and function of AER
  • induces the expression of gremlin
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19
Q

What is the function of gremlin?

A
  • inhibits BMP- which inhibits FGF-4 in AER

- inhibits Gli-3 in posterior part of limb bud

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20
Q

What is the function of signaling molecule Gli-3 (in anterior part of limb bud)?

A

inhibits the expression of shh in anterior part of the limb bud.

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21
Q

What are the advantages of using chick embryos in limb development research?

A
  • avian limbs are usually larger than amphibian limbs
  • avian embryonic limbs are accessible even though they are enclosed within extraembryonic membranes
  • because the chick embryo usually lies on its left side, the right limb bud can be used in experimental manipulations while the left limb remains untouched as the control
  • a limb bud from a donor embryo may be completely removed and transplanted to the CAM of a host embryo where it will continue to develop.
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22
Q

What is the AER?

A

apical ectodermal ridge, it is the limb ectoderm in birds.

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23
Q

What is the location of AER? What does the dorsal express? What does the ventral express?

A
  • located at the border between dorsal ectoderm and ventral ectoderm
  • dorsal expresses radical fringe
  • ventral expresses engrailed-1 (En-1)
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24
Q

What is the role of the AER?

A

interacts with underlying limb bud mesoderm and promotes continued outgrowth of limb bud

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25
Q

What are the outgrowth promoting signals?

A

FGFs (8, 4, 9, 17), and Wnts

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26
Q

Cells in the posterior part of the limb bud are under the influence of what signaling molecule and differentiate into proximal limb components?

A

retinoic acid

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27
Q

What is the role of limb bud mesoderm?

A

involved in the determination of actual limb morphology

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28
Q

What is the result if the AER is removed?

A

cessation of further outgrowth of the limb bud

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29
Q

Cells in the distal limb bud express what?

A

Msx-1

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30
Q

Hoxd gene expression in chick limb bud is involved in pattern formation in what axis?

A

proximodistal limb axis

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31
Q

What is a stylopod?

A

single proximal element, forms humerus or femur

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32
Q

What is a proximal zeugopod?

A

two parallel elements, form radius/ulna or tibia/fibula

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33
Q

What is a distal zeugopod?

A

twelve elements, form carpals or tarsals

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34
Q

What is a proximal autopod?

A

five elements, form metacarpals/metatarsals

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35
Q

What is a distal autopod?

A

Variable number of elements, form phalanges

36
Q

The mesenchyme portion of the limb bud is determined early - before closure of the neural tube. If the limb bud mesenchyme is transplanted under the epidermis of the flank or head, what is the result?

A

a limb will form

37
Q

The structure of the developing limb in every case us determined by the nature of the mesodermal components. Experimentally, it is possible to combine the leg bud mesoderm with what of a wing bud and vice versa?

A

epidermis

38
Q

Effect of limb mesoderm on overlying ectoderm is an example of what?

A

instructive induction. Normal wing bud mesoderm combined with mutant AER ectoderm fails to produce an AER.

39
Q

What occurs if the AER is removed from a chick wing bud?

A

further outgrowth of the limb mesoderm ceases

40
Q

What is the result if extra AER is grafted over limb mesoderm?

A

additional limbs form

41
Q

Mesoderm produces what that maintains the AER in a healthy state?

A

apical ectodermal maintenance factor (AEMF)

42
Q

AER maintains the mesodermal cells below it in a labile and mitotic state. What does this allow to happen?

A

Limb bud continues to grow distally, and various regions of the limb begin to appear in a proximal-distal direction.

43
Q

Necrotic zones of programmed cell death development within the limb bud ________ and will “carve” that shape of the limb?

A

mesoderm

44
Q

If cells from the potential necrotic zone are removed and cultured in vitro, the cells will die on cue. This suggests that apoptosis is a mechanism of what?

A

morphogenesis.

45
Q

What does the posterior necrotic zone form?

A

area that will become the axilla

46
Q

The limb bud mesenchyme consists of cells derived from where?

A

lateral plate mesoderm

47
Q

Limb bud mesenchyme gives rise to what?

A

appendicular skeletal elements, connective tissue, and some blood vessels.
NOTE: mesenchymal cells destined to form limb musculature will migrate into the limb bud from the somitic ventral dermomyotomes.

48
Q

Initial mesenchymal condensations for the appendicular skeleton form in what part of the limb bud, and these cells are restricted to the formation of what?

A

proximal part of the limb bud; cartilage

49
Q

Cartilage formation is restricted to the central core of the limb bud due to inhibitory restrains caused by the overlying limb bud ectoderm, which produces what?

A

wnt7a

50
Q

What does mesenchymal pre-cartilage condensations express?

A

BMP-2 and BMP-4

51
Q

What is expressed in later cartilage?

A

BMP3

52
Q

To what cells are BMPs later restricted to?

A

perichondrial cells

53
Q

Cartilage cells being to ________ and form a whorled pattern when seen in cross-section.

A

hypertrophy

54
Q

Hypertrophic cells express __________ which may induce expression of __________.

A

indian hedgehog (ihh); BMP-6

55
Q

noggin inhibits what TF?

A

BMP

56
Q

BMP is necessary to maintain what?

A

cartilage development

57
Q

What forms as a continuous precartilaginous rods?

A

limb skeleton

58
Q

What traverses the rod at the point of future joint formation?

A

condensed zone of cells

59
Q

What fills the future joint area?

A

loose connective tissue

60
Q

What forms the articular cartilages?

A

The skeletal elements on either side of the joint cavity.

61
Q

What does mesenchyme cell condensations form?

A

Other joint components

62
Q

Proximal cells of the limb bud produce what?

A

scatter factor

63
Q

Pre-muscle cells in the somite express what?

A

C-met, the receptor for scatter factor

64
Q

What do migrating pre-muscle cells express?

A

Pax 3 and N-cadherin

65
Q

Differentiation of premuscle cells into muscle within the limb bud depend on what expressed by the limb bud ectoderm?

A

wnt-6

66
Q

After skeletal element condensations form, myogenic cells condense into two muscle masses. What are they?

A

mass of flexor muscles and mass of extensor muscles

67
Q

Tcf-4 is expressed by what?

A

connective tissue associated with the muscle masses

68
Q

What determines the morphology of individual muscle?

A

connective tissue

69
Q

Muscle growth occurs via addition of materials from what cells?

A

satellite cells

70
Q

Before entering the limbs, the nerves become interconnected and form what?

A

networks or plexuses

71
Q

What are the major plexuses in higher vertebrates?

A

cervical, brachial, and lumbar plexuses

72
Q

Motor axons originate from the spinal cord during what week?

A

5th week

73
Q

Motor axons grow into limb ______ and ______ muscle masses

A

dorsal; ventral

74
Q

Motor axons originating from more medial locations in the spinal cord grow into what muscle mass?
A. medial
B. dorsal
C. ventral

A

ventral

75
Q

Motor axons originating from more lateral locations in the spinal cord grow into what muscle mass?
A. medial
B. dorsal
C. ventral

A

dorsal

76
Q

Spinal nerves supplying the limbs in terrestrial animals are _________ than other spinal nerves and the corresponding ganglia are _________.

A

larger; larger

77
Q

What is the outcome if the salamander forelimb fails to develop?

A
  • nerves of the brachial plexus remain smaller/thinner than they would if a limb had been there.
  • spinal ganglia (3, 4, 5) associated with brachial plexus, are also smaller
78
Q

If an additional limb rudiment is transplanted to the flank of the salamander embryo, what is the outcome?

A
  • local spinal nerves supply the innervation to the transplanted limb limb and these local nerves increase thickness
  • ganglia associated with these nerves increase in size; the increase in number of cells may be as much as 40%
79
Q

What is the outcome if the forelimb rudiment of an amphibian embryo, prior to outgrowth of nerves, is cut out and transplanted to a position very near the original one?

A
  • the brachial nerves will derivate from their normal paths and will be deflected in the direction of those transplanted limbs.
  • If the distance of the transplanted limb from the original position is not too great, the brachial nerves will penetrate into the limb and ramify; the limb will be fully functional.
80
Q

What is the outcome if an additional limb is transplanted into the immediate vicinity of the host limb?

A

the brachial nerves will develop branches running out to the additional limb and will supply it.

81
Q

What is the outcome of the normal path of the nerves is blocked by some obstacle?

A

the outgrowing nerves may avoid the obstacle, go around it, but ultimately will reach their normal destination.

82
Q

What is the outcome if the rudiment transplant is placed further away from the normal limb or obstacle too great?

A

nerve fibers fail to be attracted to the limb, but it may attract local nerves. limb will be non-functional

83
Q

If limbs are transplanted to the head, what happens

A

CN innervate, but are non-functional.

84
Q

If an eye was transplanted onto the side of an embryo after the forelimb rudiment was removed, what would be the outcome?

A

nerves would grow towards an eye, but would fail to penetrate and establish a connection with it.

85
Q

Early vascularization arises from endothelial cells arising from where?

A

aorta, cardinal veins, and angioblasts.

86
Q

What germ layer inhibits formation of blood vessels?

A

ectoderm