The conversion of mesenchyme to epithelium during the embryonic development of the mammalian kidney requires reciprocal inductive interactions between the ureter and the responding metanephric mesenchyme. The Pax-2 gene is activated in the mesenchyme in response to induction and is subsequently down-regulated in more differentiated cells derived from the mesenchyme. Pax-2 belongs to a family of genes, at least three of which encode morphogenetic regulatory transcription factors. In order to determine the role of Pax-2 during kidney development, we have generated a loss- of-function phenotype using antisense oligonucleotides in mouse kidney organ cultures. These oligonucleotides can specifically inhibit Pax-2 protein accumulation in kidney mesenchyme cells, where the intracellular concentrations are maximal. The kidney organ cultures were stained with uvomurulin and laminin antibodies as markers for epithelium formation. With significantly reduced Pax-2 protein levels, kidney mesenchyme cells fail to aggregate and do not undergo the sequential morphological changes characteristic of epithelial cell formation. The data demonstrate that Pax-2 function is required for the earliest phase of mesenchyme-to-epithelium conversion.
REFERENCES
Adams
B.
, Doerfler
P.
, Aguzzi
A.
, Kozmik
Z.
, Urbanek
P.
, Maurer-Fogy
I.
, Busslinger
M.
(1992
). Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and the adult testis.
Genes Dev
6
, 1589
–1607
Agrawal
S.
, Goodchild
J.
, Civeira
M. P.
, Sarin
P. S.
, Zamecnik
P. C.
(1989
). Phosphoroamidate, phosphorothioate, and methylphosphonate analogs of oligodeoxynucleotides: inhibitors of replication of human immunodeficiency virus.
Nucleosides Nucleotides
8
, 819
–823
Agrawal
S.
, Temsamani
J.
, Tang
J. Y.
(1991
). Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice.
Proc. Natl. Acad. Sci. USA
88
, 7595
–7599
Baldwin
C. T.
, Hoth
C. F.
, Amos
J. A.
, da-Silva
E. O.
, Milunsky
A.
(1992
). An exonic mutation in the HuP2 paired domain gene causes Waardenburg's syndrome.
Nature
355
, 637
–638
Balling
R.
, Deutsch
U.
, Gruss
P.
(1988
). undulated, a mutation affecting the development of the mouse skeleton, has a point mutation in the paired box of Pax1.
Cell
55
, 531
–535
Chang
E. H.
, Yu
Z.
, Shinozuka
K.
, Zon
G.
, Wilson
W. D.
, Strekowska
A.
(1989
). Comparative inhibition of ras p21 protein synthesis with phosphorus-modified antisense oligonucleotides.
Anticancer Drug Des
4
, 221
–232
Deutsch
U.
, Dressler
G. R.
, Gruss
P.
(1988
). Pax1, a member of a paired box homologous murine gene family is expressed in segmented structures during development.
Cell
53
, 617
–625
Dressler
G. R.
, Deutsch
U.
, Chowdhury
K.
, Nornes
H. O.
, Gruss
P.
(1990
). Pax2, a new murine paired-box containing gene and its expression in the developing excretory system.
Development
109
, 787
–795
Dressler
G. R.
, Douglass
E. C.
(1992
). Pax-2 is a DNA-binding protein expressed in embryonic kidney and Wilms' tumor.
Proc. Natl. Acad. Sci. USA
89
, 1179
–1183
Dressler
G. R.
, Wilkinson
J. E.
, Rothenpieler
U. W.
, Patterson
L. T.
, Williams-Simon
L.
, Westphal
H.
(1993
). Deregulation of Pax-2 expression in transgenic mice generates severe kidney abnormalitites.
Nature
362
, 65
–67
Ekblom
P.
(1989
). Develomentally regulated conversion of mesenchyme to epithelium.
FASEB J
3
, 2141
–2150
Ekblom
M.
, Klein
G.
, Mugrauer
G.
, Fecker
L.
, Deutzmann
R.
, Timpl
R.
, Ekblom
P.
(1990
). Transient and locally restricted expression of Laminin A chain mRNA by developing epithelial cells during kidney organogenesis.
Cell
60
, 337
–346
Epstein
D. J.
, Vekemans
M.
, Gros
P.
(1991
). splotch (Sp2H), a mutation affecting development of the mouse neural tube, shows a deletion within the paired homeodomain of Pax3.
Cell
67
, 767
–774
Grobstein
C.
(1953
). Morphogenetic interaction between embryonic mouse tissues separated by a membrane filter.
Nature
172
, 869
–871
Grobstein
C.
(1956
). Trans-filter induction of tubules in mouse metanephrogenic mesenchyme.
Exp. Cell Res
10
, 424
–440
Grueneberg
H.
(1954
). Genetical stidies on the skeleton of the mouse. XII. The development of undulated.
J. Genetics
52
, 441
–455
Herzlinger
D.
, Koseki
C.
, Mikawa
T.
, Al-Awqati
Q.
(1992
). Metanephric mesenchyme contains multipotent stem cells whose fate is restricted after induction.
Development
114
, 565
–572
Hill
R. E.
, Favor
J.
, Hogan
B. L. M.
, Ton
C. C. T.
, Saunders
G. F.
, Hanson
I. M.
, Prosser
J.
, Jordan
T.
, Hastie
N. D.
, van Heyningen
V.
(1991
). Mouse small eye results from mutations in a paired-like homeobox-containing gene.
Nature
354
, 522
–525
Jokelainen
P.
(1963
). An electron microscopic study of the early development of the rat metanephric nephron.
Acta Anat. Suppl
47
, 1
–73
Klein
G.
, Langegger
R.
, Timpl
R.
, Ekblom
P.
(1988
). Role of laminin A chain in the development of epithelial cell polarity.
Cell
55
, 331
–341
Klein
G.
, Langegger
M.
, Goridis
C.
, Ekblom
P.
(1988
). Neural cell adhesion molecules during embryonic induction and development of the kidney.
Development
102
, 749
–761
Koseki
C.
, Herzlinger
D.
, Al-Awqati
Q.
(1992
). Apoptosis in metanephric development.
J. Cell Biol
119
, 1327
–1333
Neckers
L.
, Whitesell
L.
, Rosolen
A.
, Geselowitz
D. A.
(1992
). Antisense inhibition of oncogene expression.
Crit. Rev. Oncogen
3
, 175
–231
Panayotou
G.
, End
P.
, Aumailley
M.
, Timpl
R.
, Engel
J.
(1989
). Domains of laminin with growth factor activity.
Cell
56
, 93
–101
Phelps
D. E.
, Dressler
G. R.
(1993
). Aberrant expression of Pax-2 in Danforth”s short tail (Sd) mice.
Dev. Biol
157
, 251
–258
Pueschel
A.
, Westerfield
M.
, Dressler
G. R.
(1992
). Comparative analysis of Pax-2 protein distributions during neurulation in mice and zebrafish.
Mech. Dev
38
, 197
–208
Sariola
H.
, Saarma
M.
, Sainio
K.
, Arumae
U.
, Palgi
J.
, Vaahtokari
A.
, Thesleff
I.
, Karavanov
A.
(1991
). Dependence of kidney morphogenesis on the expression of nerve growth factor receptor.
Science
254
, 571
–573
Stanton
B. R.
, Perkins
A. S.
, Tessarollo
L.
, Sassoon
D. A.
, Parada
L. F.
(1992
). Loss of N- myc function results in embryonic lethality and failure of the epithelial component of the embryo to develop.
Genes Dev
6
, 2235
–2247
Takahashi
Y.
, Nogawa
H.
(1991
). Branching morphogenesis of mouse salivary epithelium in basement membrane-like substratum separated from mesenchyme by the membrane filter.
Development
111
, 327
–335
Tassabehji
M.
, Read
A. P.
, Newton
V. E.
, Harris
R.
, Balling
R.
, Gruss
P.
, Strachan
T.
(1992
). Waardenburg's syndrome patients have mutations in the human homologue of the Pax-3 paired box gene.
Nature
355
, 635
–636
Ton
C. C. T.
, Hirvonen
H.
, Miwa
H.
, Well
M. M.
, Monagghan
P.
, Jordan
T.
, van Heyningen
V.
, Hastie
N. D.
, Meijers-Heijboer
H.
, Drechsler
M.
, Royer-Pokora
B.
, Collins
F.
, Swaroop
A.
, Strong
L.C.
, Saunders
G. F.
(1991
). Positional cloning and characterization of a paired box and homeobox-containing gene from the aniridia region.
Cell
67
, 1059
–1074
Vestweber
D.
, Kemler
R.
, Ekblom
P.
(1985
). Cell adhesion molecule uvomorulin during kidney development.
Dev. Biol
112
, 213
–221
Walther
C.
, Guenet
J.-L.
, Simon
D.
, Deutsch
U.
, Jostes
B.
, Goulding
M.
, Plachov
D.
, Balling
R.
, Gruss
P.
(1991
). Pax: a multigene family of paired box-containing genes.
Genomics
11
, 424
–434
Woolf
T. M.
, Melton
D. A.
, Jennings
C. B. G.
(1992
). Specificity of antisense oligonucleotides in vivo.
Proc. Natl. Acad. Sci. USA
89
, 7305
–7309
This content is only available via PDF.
© 1993 by Company of Biologists
1993
-HUBSwebinar.jpg?versionId=4486)
(update)-InPreprints.png?versionId=4486)
