Friday, April 6, 2012

UPD Bio 12 Lab Frog Development


Frog Development

1.  Fertilization

2.  Cleavage
·     egg is divided into progressively smaller units
·     blastomeres to cells

3.  Gastrulation
·     completed with formation of mesoderm and notochord

2 Primary Germ Layers

a.     Epiblast
à presumptive ectoderm
à mesoderm
b.     Endoderm

Archenteron

·     formed from integrated movements of sheets of cells
·     primary embroynic gut cavity

Blastopore

·     opening into archenteron
·     located approximately in region of posterior end of gut cavity/anus

Notochord

·     comes from epiblast
·     axis around which vertebral column will be built

4.  Organogenesis (Neurulation)
·     embryo starts to elongate dorsal thickening of ectoderm à neural or medullary plate
·     gives rise to entire nervous system
·     closes over dorsally to form neural and brain cavities
·     acquisition of characteristic shape of tadpole

5.  External Gill Stage
·     hatches from jelly capsule
·     known as larva/tadpole
·     organ systems are not developed to the adult form until metamorphosis is achieved
·     external gills
à respiration
à function until replaced by internal gills

6.  Metamorphosis
·     change in structure and function of different parts of the body
·     ceases to feed
·     lose outer skin, horny jaw, and frilled lips
·     mouth changes from small oval suctorial organ to wide slit with enlarged tongue
·     intestine shortens and changes histologically
·     stomach and liver enlarge
·     from herbivorous to carnivorous
·     respiratory organ from external gills to lungs
·     tail regression
·     sex differentiation
7.  Adult

THE MALE FROG


Distinguishing Characteristics of Male Frog
1.  smaller
2.  darkened thumb pad
3.  low, guttural croaking sound
4.  slender and streamlined body
5.  absence of coelomic cilia except in kidneys

Mesorchium
-     double fold of peritoneum
-     suspends testis to kidneys
-     surrounds each testis
-     continuous with peritoneum epithelium

Spermatogenesis
-     seasonal
-     completed within the testis
-     Meiotic arrests (?)

1.  Spermatogonium
·     after breeding season à frog ceases all metabolic activity
·     enters period of rest
·     granular, oval nucleus
·     found near basement membrane of seminiferous tubules
·     small, closely packed cells

2.  Primary Spermatocyte
·     large cells
·     irregularly spherical
·     large and vesicular nuclei
·     extended prophase à nucleus undergoes series of changes

a.  Leptotene
à cytoplasm and nucleus grow
b.  Zygotene
c.  Pachytene
d.  Diplo-diakinesis
à homologous chromosomes start to separate
à chromatin material, volume of nucleus and cytoplasm increases
à nuclear membrane breaks down

3.  Secondary Spermatocyte
·     half the size of primary spermatocyte
·     lie towards the lumen
·     darkly staining nucleus
·     cytoplasm may be tapered on one side

4.  Spermatid
·     smaller
·     condensed nucleus of irregular shape
·     clusters of spermatids appear as cluster of granules

5.  Spermatozoon
·     Elongated, solid-staining head (nucleus)
·     Has acrosome

Sertoli Cells

-     Relatively large, columnar-type basal cell
-     Where head of spermatozoon converge
-     Functionally a nurse cell à supply nutrition to spermatozoon

THE FEMALE FROG


Distinguishing Characteristics of the Female Frog
1.  generally larger
2.  absence of dark thumb pad
3.  inability to produce lateral cheek pouches from croaking
4.  flabby and distended abdomen
5.  presence of peritoneal cilia

The Ovary
Mesovarium
-     2-layered
-     continuous with theca externa

Theca Interna
-     “cyst wall”
-     contains smooth muscle fibers
-     derived from retro-peritoneal tissue

Ovarian Follicle
1.  theca externa
2.  theca interna
3.  follicle cells
·     oval and granular nucleus
·     derived from oogonia
·     surround developing oocyte

The Egg
1.  Animal Pole
a.     cytoplasm
b.     germinal vesicle
2.  Vegetal Pole à Yolk

Vitelline Membrane
-     enclosed within follicle cells
-     closely applied to mature egg
-     non-cellular and transparent
-     sometimes designated as primary of several egg membranes

Jelly Coats
-     “albumen”


Layers
-     largely water
-     greater than diameter of egg
-     (Inner to outer)
1.  thin, heavy consistency
2.  thin, more fluid
3.  greater in thickness, less viscous

Oogenesis
-     after breeding season à only oogonia
·     no pigment
·     little or no yolk
·     represents an ovarian unit
à many follicle cells
à 1 ovum

Process of Egg Maturation
1.  chromatin nucleoli aid in synthesis of yolk
2.  breakdon of germinal vesicle à intermingling of cytoplasmic and nuclear components

Growth Period to Primary Oocyte Stage
-     achieved by accumulation of yolk

1.     cells no longer divide by mitosis and is known as an oocyte rather than oogonia
2.     yolk nuclei
·     granules within cytoplasm
·     function as centers of yolk accumulation
·     source of yolk: digested food of female
3.     accumulation of yolk à concentrated in one side of oocyte

First Indicators of Egg Polarity
1.  growth changes
2.  unequal distribution of pigment, yolk, cytoplasm

Established Polarity of the Frog’s Telocithal Egg
1.  Animal Hemisphere
a.  cytoplasm
b.  melanin
c.  nucleus
2.  Vegetal à Yolk

Surface layer of amphibian egg
-     contains yolk grains and black pigment granules
-     with each cleavage, the superficial coat is divided between blastomeres

Vitelline Membrane at Growth Period
1.  thin
2.  transparent
3.  nonliving
4.  closely adherent

Ovulation and Maturation
-     brought about by LH
-     liberation of egg from ovary
-     rupture and emergence of eggs from individual follicles
-     theca externa ruptures

Maturation Divisions
1.  First maturation division
·     occurs during ovulation

a.  primary oocyte
b.  first polar body
à small amount of cytoplasm
à no yolk
2.  Second maturation division
·     begins without any intermediate rest period
·     occurs around the time the egg enters the oviduct
·     egg reaches uterus arrested at Metaphase II
·     maturation is not completed until egg is activated by sperm

a.     secondary oocyte
b.     second polar body

CLEAVAGE

Egg: Telocithal
à large amount of yolk concentrated at one pole
à yolk is opposite to the concentration of cytoplasm and location of nucleus

Cleavage: Holoblastic

Blastomeres
-     cell of the embryo (from 1st cleavage to late blastula)

1.     micromeres à upper, smaller
2.     macromeres  à lower, larger

BLASTULATION
-     segmentation of the egg à division into smaller cellular units
-     appearance of internal cavity à blastocoel

Blastula
-     stage of development where blastocoel is present
-     overlaps stages of cleavage
-     coeloblastula

Blastocoel
-     filled with albuminous fluid from surrounding cells
-     eccentric position
·     above level of equator
·     slightly toward gray crescent side of cleaving egg
-     size increases with formation of smaller and smaller surrounding cells
-     later displaced by archenteron

Gradient of Cleavage
-     active region à animal pole
-     least active region à vegetal pole
Changes in Embryo
1.  actual migration of small and pigmented ccells toward vegetal pole à thinning of roof of blastocoel

2.  outermost layer of cells
a.  single outer layer of cells
à contains most of superficial pigments
à recognized as epidermal layer
à gives rise to epithelium
Ø  integument
Ø  lining of nervous system
b.  inner tiers of cells of blastular roof
à less pigmented
à collectively known as nervous layer
à give rise largely to neuroblasts of nervous system

Germ Ring
-     where yolk is most actively being transformed into cytoplasm
-     has something to do with subsequent formation of lips of future blastopore

GASTRULATION
-     dynamic process
-     results in transformation of single layered blastula to two-layered embryo
-     involves but is independent of mitosis

Germ Layers
1.  ectoderm
2.  ectoblast
·     outermost layer of blastula
a.     mesoderm
b.     endoderm

-     first distinguishable with gastrulation
-     definable by fate in embryonic devt

Basis for Distinction of Germ Layers
1.  position in developing organism
2.  fate

Importance of Gastrulation
1.  cell division
2.  cell differentiation
3.  morphogenetic movements
-     independence and interdependence of various areas

Embryos at Gastrulation
                Senstive to…
1.  changes in physical environment
2.  genetic incompatibilities within chromosomes

Morphogenetic Movements

1.  Invagination
·     Insinking of the egg surface followed by forward migration which involves displacement of inner materials

2.  Involution
·     turning inward
·     a rotation of material upon itself so that movement is directed toward the interior of the egg

3.  Epiboly
·     progressive extension of cortical layer of animal hemisphere toward vegetal hemisphere, or,
·     the expansion from animal to vegetal hemisphere

Gastrulation Proper
1.  thinning of gray crescent side of blastula wall
·     migration of deeper layers of blastula away from original site of gray crescent
·     region where involution first occurs
·     eventually known as dorsal lip

2.  continued epiboly toward vegetal hemisphere so that diameter of marginal zone becomes progressively smaller as it passes below the equator
·     marginal zone à high rigidity à inward pressure on yolk cells à arch upward toward blastocoel
·     yolk endoderm cells toward gray crescent side are forced upward à reduction in blastocoel size

3.  initial involution at lower margin of original gray crescent
·     lateral extension of this involution along epibolic marginal zone
·     initial involution dorsal lip

4.  further epiboly of entire marginal zone toward vegetal hemisphere

5.  piling up of animal hemisphere cells
·     many are destined to move inside over blastoporal lips

6.  continued lateral extension of involuting marginal zone
·     circumferential meeting of blastoporal lips
·     yolk plug

7.  reduction in size of circumblastoporal lips and exposed yolk plug

8.  the origin of endoderm à arising from involuted dorsal lip cells
·     endoderm fans out to give rise to archenteron

9.  simultaneous with origin of endoderm, some cellsa re proliferated between roof of archenteron and dorsal epiblast à notochord
chordamesoderm à before differentiation

-     involuted cells extend anteriorly
-     inturned endoderm surrounds archenteron
-     archenteron expands
-     blastocoel becomes progressively smaller
-     involuting cells + expanding endoderm à enlarged archenteron lined with endoderm
-     Blastopore
·     opens into archenteron
·     occluded by yolk endoderm cells
-     overlying ectoderm thickens
-     whole embryo elongates
-     start of neurulation

NEURULATION AND EARLY ORGANOGENY

Factors Inducing Ectodermal Thickening
1.  notochord
2.  roof of archenteron

Neural Plate (Medullary Plate)
-     ectodermal thickening
-     extends from dorsal lip of blastopore in an anterior direction
-     median band of thickened ectoderm
-     where the brain will develop

Beginning of Central Nervous System Formation
1.  lateral folds
2.  transverse neural fold
3.  neural groove

Surface Changes
-     shape becomes elongate
-     neural plate provides slightly elevated (convex) dorsal surface à becomes flattened à concave
-     distinct head
-     ovoid body à due to yolk

Causal Factor in Change in Shape of Embryo
-     appearance of thickened and elongating neural plate
-     subsequent formation of neural tube

Neurula Stage
-     nervous structures are being formed

1.  transverse neural fold
·     anterior junction of folds
·     represents anterior extremity of developing brain
2.  lateral neural folds
·     paired
·     extensive
·     more posterior
·     represent posterior parts of the brain and spinal cord levels
·     more toward each other

Neural Tube
-     neurocoel
-     fusion occurs last at the extremities
-     enclosed canal lined with ciliated and pigmented epithelium
-     found as the central canal in brain and spinal cord of adult

Sense Plates
-     elevated ridge of ectoderm
-     gives rise to
1.  mandibular arches
2.  lens of the eye
3.  nasal placodes
4.  oral suckers

Stomodeal Cleft
-     ventral groove
-     sperated by two mandibular ridges

Oral Suckers
-     adhesive
-     mucus-secreting glands

Stomodeum
-     ectodermal
-     mouth
-     part of mouth derived from stomodeum will be lined with ectoderm

Proctodeum
-     invaginating pit ventral to blastopore
-     ectodermal
-     primordium of anus
-     establishes a new ectodermally lined opening into hindgut

Dorsal Fin
-     developed initially by posterior growth of myotomes

Ependymal Layer
-     lining of central canal
-     ciliated and pigmented cells

Nervous Ectoderm
-     nervous layer of ectoderm of blastular roof
-     gives rise to neuroblasts of CNS

Neural Crest
-     nervous layer at level of dorsal fusion is pinched off
-     part of original ectodermal neural folds which don’t form an integral part of neural tube
-     paired
-     extend the full length of CNS
-     give rise to

1.  ganglia of CNS
2.  ganglia of sympathetic nervous system
3.  chromatophores

Level of Brain
1.  ganglia of CNS
2.  ganglia of sympathetic nervous system
3.  chromatophores
4.  visceral cartilage
5.  cranial cartilage

Body Level
1.  paired spinal ganglia
2.  sympathetic nervous system
3.  chromatophores
4.  medulla of adrenal gland

SURVEY OF MAJOR DEVELOPMENTAL CHANGES

Primary Parts of Embryonic Brain
1.  Prosencephalon à forebrain
2.  Mesencephalon à midbrain
3.  Rhombencephalon à hindbrain

Prosencephalon
-     primary forebrain
-     develops almost immediately
-     gives rise to optic vesicles

NOTE: prosencephalon à optic vesicles à (walls) lens and cornea


Derivatives of Forebrain
1.  infundibulum
·     ventral to notochord
·     vesicular outpocketing of floor of prosencephalon
·     along with epithelial hypophysis à posterior pituitary (?)
·     hypophysis
à pigmented ectodermal cells
à between infundibulum and roof of pharynx
2.  optic chiasma
·     floor of forebrain
·     thickening
·     crossing-over
3.  optic recess
·     depression associated with lateral optic vesicles
·     anterior to optic chiasma
·     connected with optic stalk
4.  epiphysis
·     roof of prosencephalon
·     evagination of roof
·     forerunner of pineal body
5.  anterior choroid plexus
·     anterior to epiphysis
·     non-nervous
·     vascular
·     folded
6.  optic vesicles
·     paired primordia of eye
·     paired lateral vesicular evaginations of forebrain

7.  olfactory placodes
·     paired
·     button-like thickening of pigmented surface ectoderm

Rhombencephalon
1.  auditory placodes

The Enteron (Gut Cavity)
1.  Foregut
2.  Midgut
3.  Hindgut

Foregut
1.  oral evagination
·     evagination of endoderm
·     oral evagination + stomodeal invagination à oral plate à mouth
2.  pharynx
·     large cavity of foregut
·     expands laterally to form endodermally lined visceral pouches
3.  liver diverticulum
·     forerunner of
a.     bile duct
b.     gall bladder
c.     liver

Axial Skeleton
1.  notochord
·     mesodermal origin
·     expands and becomes vacuolated

Mesoderm and Its Derivatives

Mesenchyme
-     embryonic, loosely dispersed presumptive mesoderm
-     most anterior mesoderm is in this form
a.     head
b.     pharyngeal region

Arches
-     vertical concentrations of mesoderm

Somites
-     mesoderm posterior to pharynx and lateral to notochord
-     inverted horseshoe shape around archenteron and yolk

1.  Epimere à Mesodermal Somite
·     posterior to pharynx
a.     dermatome à cutis plate
à dermis
à appendage musculature
b.     myotome
à skeletal muscles of back
à and body
c.     sclerotome
à mesenchymal
à between myotome and notochord
à vertebral skeleton
2.  Mesomere à Intermediate Mesoderm
a.  Excretory System
Mesomere à nephrotome à nephrocoel à pronephros
b.  Urogenital Tract
3.  Hypomere à Lateral Plate Mesoderm
a.  Somatic
à somatic + ectoderm = somatopleure à skin, its blood and CT
à pericardial membrane
b.  Splanchnic
à splanchnic + gut endoderm = splanchnopleure
Ø  lining of epithelium
Ø  muscles
Ø  blood vessels of midgut
Ø  blood vessels of hindgut
à myocardium
c.  Coelom
à pleuro-peritoneal cavity
à pericardial cavity

7 mm TADPOLE

INVAGINATION

of
Anterior choroid plexus
Roof of prosencephalon
External nares
Epidermal
Optic cup
Optic vesicle
Otic vesicle
Epidermal ectoderm
External ear
ectoderm

EVAGINATION

of
Infundibulum
Diencephalon
Ventral pancreas
Duodenum
Epiphysis
Prosencephalon
Optic recess
Median ventral
Optic vesicle
Forebrain

Mesenchymal Cells

1.     dermatome
2.     sclerotome

Condensations of Mesenchyme

1.     larval brain case
2.     visceral arches

Brain Divisions

1.     Prosencephalon
2.     Mesencephalon
3.     Rhombencephalon

Structures Associated with Prosencephalon

1.     prosocoel
2.     epiphysis
3.     anterior choroid plexus
4.     future telencephalon
5.     future diencephalon
6.     external nares – olfactory pits
7.     olfactory canal
8.     olfactory epithelium
9.     internal naris
10.   olfactory nerve

Structures Associated with Diencephalon

1.     optic vesicles
a.     optic cup
b.     optic lens
c.     retina
c.1     rods
c.2     cones
d.     pigmented epithelium
2.     optic recess
3.     optic chiasma
4.     optic nerve
a.     stalk
b.     ganglion cells
c.     bipolar neurons

Structures Associated with Mesencephalon

1.     infundibulum
2.     hypophysis
3.     mesocoel
4.     trigeminal ganglion

Structures Associated with Rhombencephalon

1.     rhombocoel (4th ventricle)
2.     basal plates
3.     posterior choroid plexus
4.     otic vesicles
5.     acoustico-facialis
a.     facial (VII)
b.     auditory (VIII)
6.     endolymphatic duct
7.     glossopharyngeal ganglion (IX)

Spinal Cord

1.     central canal
2.     ependymal layer
3.     mantle layer
4.     marginal layer

Digestive System

                Anterior to posterior

1.     oral cavity
2.     pharynx – up to external gill section
3.     esophagus
a.     esophageal plug
4.     stomach
5.     duodenum
6.     midgut
7.     hindgut
8.     cloaca
9.     proctodeum – anus

Mesodermal Derivatives

1.     bulk of skeleton and muscle
2.     entire circulatory system
3.     epithelium lining of body cavity

Blood Tracing
      *starting from the heart*

Arterial System: Pulmonary Circuit

1.     conus arteriosus
2.     truncus arteriosus
3.     afferent branchial vessels of the aortic arches
4.     gill capillaries
*efferent branchial vessels* à vein(?)

Arterial System: Systemic Circuit

1.     conus arteriosus
2.     truncus arteriosus
3.     afferent branchial vessels of the aortic arches
4.     dorsal aortae
5.     dorsal aorta (merging at level of stomach)
6.     caudal artery (after cloaca)

Venous System: Draining the head part

1.     sinus venosus
2.     common cardinal vein (ducts of Cuvier)
3.     anterior cardinal vein

Venous System: Draining the posterior part

1.     sinus venosus
2.     common cardinal veins
3.     posterior cardinal veins
4.     posterior cardinal vein (merge at level of cloaca)
5.     caudal vein (anus)

SUPER DOOPER MEGA TABLE

Ectodermal Derivatives

Neural Tube

1.     Prosencephalon
a.     future telen
b.     future diencephalon
2.     mesencephalon
3.     rhombencephalon
4.     anterior choroid plexus
5.     epiphysis
6.     infundibulum
7.     hypophysis (+outer epithelium)
8.     basal plates
9.     posterior choroid plexus
10.   spinal cord
11.   optic cup
12.   optic lens
13.   pigmented epithelium
14.   optic recess
15.   retina
16.   optic chiasma

Neurocoel

1.     prosocoel
2.     diocoel
3.     mesocoel
4.     telocoel
5.     rhombocoel
6.     central canal

Neural Folds

1.     ependymal layer
2.     mantle layer
3.     marginal layer

Neural Crest

1.     bipolar neurons
2.     ganglion cells
3.     trigeminal (V)
4.     acoustico-facialis (VII-VIII)
5.     glossopharyngeal (IX)
6.     optic (II)

Epidermal Ectoderm

1.     endolymphatic duct
2.     otic vesicle
3.     olfactory epithelium
4.     olfactory canal
5.     external nares
6.     epidermis
7.     oral suckers
8.     stomodeum depression

Mesoderm Derivatives
1.     mesenchyme
2.     germinal ridge
3.     branchial muscles
4.     notochord

Epimere (mesenchyme)
1.     dermatome
2.     myotome
3.     sclerotome
4.     branchiometric muscles
5.     parachordal cartilage
6.     trabecular cartilage
7.     visceral arches
8.     hypobranchial

Mesomere

1.     pronephros
2.     pronephric duct

Hypomere: Coelom

1.     pericardial cavity
2.     peritoneal cavity

Hypomere: Somatic

1.     parietal pericardium
2.     parietal peritoneum

Hypomere: Splanchnic

1.     Visceral pericardium
2.     Visceral peritoneum
3.     Efferent branchial vessels
4.     Afferent branchial vessels
5.     Dorsal aorta
6.     Truncus
7.     Conus
8.     Atrium
9.     Ventricle
10.   CCV
11.   SV
12.   Vitelline veins
13.   ACV
14.   PCV
15.   Caudal artery
16.   Caudal vein
17.   Gill capillaries
18.   Hepatic vein

Endodermal Derivatives
1.     laryngotracheal groove
2.     thyroid gland
3.     yolk

Archenteron

1.     pharynx
2.     pharyngeal pouches
3.     lung buds
4.     trachea
5.     esophagus
6.     stomach
7.     liver
8.     gall bladder
9.     midgut
10.   hindgut
11.   pancreas
12.   cloaca

Blastopore
1.     anus

Ectodermal Derivatives

I.              Neural Ectoderm
A.     Neural Tube
1.     Brain
2.     Spinal Cord
B.     Layers
1.     Ependymal Layer
2.     Mantle Layer à Gray matter
a.     Neuroblasts
b.     Glioblasts
3.     Marginal Layer à White matter
a.     Neurites
b.     Glial cells
II.             Epidermal Ectoderm
A.     Epidermis
B.     Epidermal Derivatives
C.     Lens
D.     Sense Organs (Invaginations)
1.     Olfactory
2.     Otic

III.            Neural Crest
A.     Level of the Brain
1.     Ganglia of CNS
2.     Ganglia of Sympathetic Nervous System
3.     Melanophores/Chromatophores
4.     Visceral Cartilage
5.     Cranial Cartilage
6.     Dura Mater
7.     Pia mater
B.     Level of the Body
1.     paired spinal ganglia
2.     sympathetic nervous system
3.     chromatophores
4.     medulla of adrenal gland

Endodermal Derivatives

I.              Gametogenic Cells (?)
II.             Trachea
III.            Lungs
IV.            Gut Appendages
A.     Pharyngeal Arches
B.     Pharyngeal Pouches
V.             Foregut
A.     Lining of esophagus
B.     Lining of stomach
C.     Anterior portion of duodenum
VI.            Midgut
A.     remaining lining of small intestine
VII.           Hindgut
A.     Large Intestine
B.     Rectum

Mesodermal Derivatives
I.              Axial Mesoderm
II.             Paraxial Mesoderm à Somites
A.     Epimere
1.     Dermatome
a.     Dermis
b.     Connective Tissue
c.     Appendage Musculature
2.     Myotome
a.     Skeletal muscles of back
b.     Skeletal muscles of body
3.     Sclerotome
a.     Vertebra
b.     Axial Skeleton
B.     Mesomere
1.     Excretory System
2.     Urogenital Tract
C.     Hypomere
1.     Somatic Layer
a.     Sompatopleure
a.1    Skin, its blood and connective tissue
b.     Pericardial membrane (parietal pericardium)
c.     Parietal Peritoneum
2.     Splanchnic Layer
a.     Splanchnopleure
a.1    Lining Epithelium
a.2    Muscles
         a.2.1    smooth
         a.2.2    myocardium (cardiac)
a.3    Blood vessels of midgut
a. 4   Blood vessels of hindgut
b.     Visceral Pericardium
c.     Visceral Peritoneum
3.     Coelom
a.     pleuro-peritoneal cavity
b.     pericardial cavity















ECTODERM

MESODERM

ENDODERM

Position

Outermost layer of cells
Middle layer of cells
Innermost layer of cells

 

 

Origin

Cells from animal hemisphere and equatorial plate cells

(Fate map: presumptive ectoderm)


Lips of blastopore


Vegetal hemisphere area

 

 

 


Derivatives


1.     epidermis
2.     epidermal structures
3.     nervous system
4.     sense organs
5.     stomodeum
6.     proctodeum
1.     skeleton
2.     connective tissue
3.     muscles
4.     blood and vascular system
5.     coelomic epithelium and derivatives
6.     urogenital system
7.     notochord
Lining epithelium of alimentary tract and its outgrowths…
a.     thyroid
b.     lungs
c.     liver
d.     pancreas
e.     etc.

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