1 Max-Planck-Institute for Biochemistry, Department of Protein Chemistry, 82152 Martinsried, Germany
2 Institute for Histology, University of Göttingen, 37075 Göttingen, Germany
3 University of Pittsburgh, Department of Neurobiology, Pittsburgh, PA 15261, USA
4 Institute for Biochemistry, University of Cologne, 50931 Cologne, Germany
5 University of Manchester, Wellcome Trust Centre for Cell-Matrix Research, Manchester M13 9PT, UK
*Author for correspondence (e-mail: ulrike.mayer{at}man.ac.uk)
Accepted 18 March 2002
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SUMMARY |
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Key words: Basement membrane, Wolffian duct, Kidney, Lung, Morphogenesis, Mouse
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INTRODUCTION |
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The precise mechanisms how basement membranes are assembled and how the biological function of the proteins is maintained within the basement membrane in vivo, are still unclear. In particular, the differential sites of synthesis for nidogen and laminin chains in mesenchymal and epithelial tissues, respectively, (Thomas and Dziadek, 1993; Ekblom et al., 1994
) argues for the presence of other factors involved in the assembly processes. Recent results obtained by targeted deletion of cell-surface receptors, such as ß1 integrins and dystroglycan showed disruption of basement membrane structures (Kreidberg et al., 1996
; Williamson et al., 1997
; DiPersio et al., 1997
; Sasaki et al., 1998
; Henry and Campbell, 1998
) and emphasize a role for transmembrane complexes in coordinating the spatial and temporal local concentrations of proteins at the sites of basement membrane formation.
It is widely believed that basement membranes serve as both structural barriers and as a substrate for cellular interactions. The genetic inactivation of most of the major components has demonstrated that each of the proteins serves specific functions. Though all mutations interfere at specific stages with basement membrane integrity, the underlying mechanisms most probably differ. Mice deficient for perlecan develop normally before they die of heart failure at 10.5 day post coitum (dpc), because of basement membrane instability caused by mechanical stress (Costell et al., 1999). Natural mutations within any of the laminin 5 chains (
3, ß2,
2) lead to junctional epidermolysis bullosa, a severe skin blistering disease (Pulkkinen and Uitto, 1999
). Likewise, mutations within the laminin
2 chain result in congenital muscular dystrophies (Helbling-Leclerc et al., 1995
). Targeted disruption of the laminin
5 chain is embryonic lethal because of local structural basement membranes defects at the site of expression of the corresponding laminin isoforms (Miner et al., 1998
). None of these mutations, however, showed major early embryonic defects, reflecting the distinct expression of specific laminin isoforms and their function during embryonic development and adulthood. On the contrary, the crucial importance of laminins for basement membrane formation has been demonstrated by deleting 10 out of the 14 known laminin isoforms through the inactivation of the laminin
1 chain. Mice, homozygous for the mutation lack basement membranes and die at 5.5 dpc through a failure of ectodermal and endodermal cell differentiation (Smyth et al., 1999
; Murray and Edgar, 2000
).
The nidogen-binding site of laminin has been localized to a single laminin-type epidermal growth factor-like (LE) module, 1III4, of the laminin
1 chain (Mayer et al., 1993b
) and is therefore present in most of the laminin isoforms known. Though the
2 chain of laminin 5 contains a highly homologous LE module,
2III4, no significant binding activity has been observed (Mayer et al., 1995
). LE motifs consist of about 60 amino acids and form four disulfide-bonded loops (Baumgartner et al., 1996
; Stetefeld et al., 1996
). Peptide and mutant analysis of the 56 amino acid nidogen-binding LE module demonstrated that two non-contiguous loops are necessary for high-affinity binding to nidogen 1 (Pöschl et al., 1994
; Pöschl et al., 1996
). Antibodies that inhibit the laminin-nidogen interaction perturbed epithelial branching morphogenesis in organ culture of lung, kidney and salivary glands, a process that involves formation of new basement membranes (Ekblom et al., 1994
; Kadoya et al., 1997
). Surprisingly, however, mice deficient for nidogen 1 failed to show any overt phenotype, which may be attributed to a redundancy of nidogen isoforms (Murshed et al., 2000
).
In order to characterize the biological significance of the laminin-nidogen interaction, we have used a genetic approach to delete the nidogen-binding module 1III4 of the laminin
1 chain in the germline of mice, thereby abrogating binding of potentially redundant nidogen isoforms. The results show that while basement membrane formation is not crucially dependent on the laminin-nidogen interaction, there are structural abnormalities in specific basement membranes of kidney and lung, resulting in impairment of early kidney organogenesis and lung development.
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MATERIALS AND METHODS |
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Generation of hetero- and homozygous ES cells
R1 cells were cultured and transfected with the NotI linearized targeting vector as described (Mayer et al., 1997). Genomic DNA from G418 resistant clones were screened by Southern hybridization after EcoRV restriction with a 0.5 kb HindIII/EcoRV fragment located upstream of the targeting vector. Positive clones were analyzed for additional random insertion using the neomycin gene as probe. Two clones were expanded and transiently transfected with Cre recombinase under the control of the PGK promoter. Deletion of the selection cassette was verified after EcoRV restriction using the same probe as described above. To obtain homozygous mutant ES cells, one of the positive clones was transfected once again with the initial targeting vector to obtain one allele still carrying the selection cassette (4.5 kb fragment) and one devoid of (2.7 kb).
Generation of mice lacking the nidogen-binding module 1III4
Two independent heterozygous clones lacking the selection cassette, CIA2 and CIII3C6, were injected into C57/B6 blastocysts and transferred into pseudopregnant CD1 foster mothers. Highly chimeric male founder mice were obtained which were crossed with C57/B6 and 129Sv females to obtain heterozygous F1 offspring. Heterozygous mice were mated to obtain time-staged homozygous embryos. All F1 and F2 progeny were genotyped by Southern blotting or by PCR using 5'-AGATGTGAACTCTGTGATGAC as forward and 5'-TGCAAGAAGTGGTTCACACCGCATTCT as reverse primer.
Preparation of embryoid bodies
Undifferentiated ES cells were trypsinized, diluted in ES medium without LIF (EB medium) as described (Wobus et al., 1991) and plated onto cell culture dishes for 45 minutes to allow residual feeder cells to attach. The supernatant was washed twice with EB medium and ES cells were finally diluted at a concentration of 32x103/ml. The cells were then placed in hanging drops of 25 µl (Wobus et al., 1991
). After 2 days, cell aggregates were transferred into EB medium filled bacterial dishes. Medium was changed every 48 hours. After 10 days in culture, intact embryoid bodies were collected, washed twice in PBS and further processed.
Protein analysis and rotary shadowing electron microscopy
Embryoid bodies were sequentially extracted with TBS and EDTA as described (Paulsson et al., 1987). The EDTA extract was passed over a Hi-Trap heparin affinity column (Pharmacia). Bound laminin was eluted with a linear NaCl gradient as previously described (Paulsson et al., 1987
). Rotary shadowing electron microscopy was performed as described (Paulsson et al., 1987
).
For radioimmuno inhibition assays (RIA) embryoid bodies from three independent homozygous mutant and control cells and embryos were extracted in RIPA buffer (Sasaki et al., 1996) and analyzed as described (Sasaki et al., 1998
). For immunoblotting, 5 µg total protein were separated under non-reducing conditions on 5-15% SDS-PAGE gels, transferred onto PDVF membranes (Millipore) and incubated with the primary antibodies. Using goat-anti-rabbit antibodies conjugated with horseradish peroxidase (BioRad), specific bands were detected after visualizing enzyme activity with ECL (Amersham).
Northern blotting and RT-PCR
Total RNA was prepared using Trizol, according to the suppliers protocol (GibcoBRL). Samples (10 µg) of each genotype were electrophoresed in a 1.2% denaturing formaldehyde gel and transferred to HybondN membrane (Amersham) and hybridized against 32P-oligolabeled cDNA probes for laminin 1, laminin
1III4, perlecan, nidogen 1 and nidogen 2. For standardization, the same membranes were reprobed with a GAPDH probe.
For RT-PCR, 5 µg of total RNA was reverse transcribed with MuLV reverse transcriptase (Appligene). For second strand synthesis 2 µl of the reaction mixture were used in PCR reactions. After 35 cycles, PCR products were analyzed on a 2% agarose gel. Control reactions were carried out without reverse transcription. Forward and reverse primers were chosen from different exons and were as follows: for demonstrating deletion of the nidogen-binding module 1III4, 5'-AGATGTGAACTCTGTGATGAC and 5'-TTGTAGTAGCCAGGGTCACAAGTA; for LE modules 6-7 of the laminin
1III domain, located downstream of the deletion site, 5'-GTGTGACTGCCATGCTTTGGG and 5'-CTTCACAAGTCGGTAACAAGCCGG; for GAPDH, 5'-CTGCCAAGTATGATGACATCA and 5'-TACTCCTTGGAGGCCATGTAG, yielding in reaction products of 369 and 201 bp for wild-type and mutated alleles for the deletion site, and 317 bp and 252 bp products for LE6-7 and GAPDH, respectively. The PCR products were verified by sequencing.
Histochemistry and immunohistochemistry
Embryos and tissues were dissected, fixed for 2-4 hours in 4% paraformaldehyde in PBS and either embedded in OCT or in paraffin wax. Paraffin wax-embedded sections (5-10 µm) were processed for Hematoxylin and Eosin staining.
For immunostaining 7-10 µm cryosections were stained with the respective antibodies. Paraffin wax-embedded sections were dewaxed and treated for 10 minutes with 100 µg/ml proteinase XXIV (Sigma) at 37°C. After a blocking step with PBS, 5% normal goat serum (NGS), first antibodies were applied in PBS, 2% NGS for 1 hour at 37°C in a humidified chamber. After washing with PBS, the sections were incubated with secondary Cy3- or Cy2-conjugated goat-anti-rabbit or goat-anti-rat antibodies. After final washing in PBS, sections were mounted and analyzed on an Axiophot fluorescence (Zeiss) and an Olympus confocal microscope. Primary antibodies used were rabbit antisera against nidogen 1 (Fox et al., 1991), nidogen 2 (Kohfeldt et al., 1998
) and laminin 1, recognizing isoforms containing
1, ß1 or
1 chains (Fox et al., 1991
), laminin
1 recognizing the wild-type or the mutated protein (Mayer et al., 1998
), perlecan, collagen IV (Costell et al., 1999
), Pax2 (Babco), SP-C (Vorbroker et al., 1995
) and rat anti-mouse monoclonal antibodies against nidogen 1 (Ries et al., 2001
).
Electron microscopy
Tissue specimens (1 mm2) were fixed in 3% paraformaldehyde and 3% glutaraldehyde in PBS for 2 hours at 4°C as described (Mayer et al., 1997). They were then postfixed for 1 hour in 1% osmium tetroxide and embedded in Epon. Ultrathin sections were collected on formvar coated copper grids, stained for 10 minutes with uranyl acetate, 5 minutes with lead citrate and examined with a Zeiss EM 109 electron microscope. For quantification of lung morphology, eight tissue samples of four homozygous mutant and four control mice were taken and a total of 200 randomly selected fields of 117 µm2 counted for each genotype.
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RESULTS |
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Kidney organogenesis is dependent on the laminin-nidogen interaction
Heterozygous mice derived from two independent ES cell lines were obtained and did not show any obvious phenotype up to one year of age. Immunostaining with antibodies specific for the interfaces of the tandem arrays of LE modules in the wild-type or mutant (Mayer et al., 1998) demonstrated that both the mutant and the wild-type laminin
1 chain were present in basement membranes (data not shown). Genotyping of more than 800 offspring of heterozygous crossings at weaning failed to reveal any surviving homozygous mutant animals (+/+:+/:/; 280:524:0). Dead pups were routinely observed shortly after birth of which most were homozygous for the mutation. This suggested that mice deficient for the nidogen-binding module
1III4 died soon after birth. However, phenotype analysis during prenatal development showed that only
60% of the mutant embryos developed to term, whereas
40% died before 11.5 dpc for so far unknown reasons.
Embryos were removed at 18.5 dpc by Caesarian section. The majority of the mutant embryos were about 20% smaller than their littermate controls. Strikingly, visual inspection demonstrated the lack of kidneys in most homozygous embryos (Fig. 3C,E) whereas they were present in all wild-type (Fig. 3A,D) and heterozygous embryos analyzed. Both kidneys were identifiable in only a small percentage of mutant embryos (Fig. 3B), while 90% showed either bilateral (80%; 46 out of 59 animals) or unilateral (10%; 5 out of 59 animals) renal agenesis, demonstrating that the laminin-nidogen interaction plays a pivotal role during kidney organogenesis. The low penetrance of metanephric development was independent of the ES clone or genetic background being observed on a heterogeneous (C57/129Sv) as well as on a 129Sv background. Despite the lack of kidneys, adrenal glands, testis and ovary were normally present with gross and histological analysis failing to reveal any abnormalities in these organs, while the uterus in female and vas deferens and the seminal vesicles in male mutants were absent (Fig. 3C,E).
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Nidogen 1 is not retained in basement membrane structures
To determine whether the deletion of 1III4 within the laminin
1 chain affects the localization of nidogen, we performed immunostaining of various tissues. Nidogen 1 was present in all basement membranes of wild-type and heterozygous animals, but, with a few exceptions, barley detectable in basement membrane structures of the mutant animals. Only basement membranes around large blood vessels and at the dermal-epidermal junction of the skin showed staining intensities comparable with the control tissues (data not shown). This raised the question as to whether the recently newly identified, highly homologous isoform, nidogen 2 (Kohfeldt et al., 1998
), is upregulated. Double-immunostaining for nidogen 1 and nidogen 2 in kidney sections, however, indicated that nidogen 2 was neither differently deposited nor was its staining intensity increased relative to the controls (Fig. 7A-D). Staining intensities similar to that of wild-type controls were also observed for the laminin
1 chain (Fig. 7E,F), collagen IV and perlecan (data not shown).
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Lung defects might cause perinatal lethality in 1III4-deficient mice
Homozygous mice die very soon after birth and, despite the fact that the kidney phenotype should lead to a perinatal lethality within the first 2 days, we have been unable to identify any living offspring within the first postnatal day. This indicated that another defect causes death of the homozygous animals. Indeed, some of the mutant dead pups had a cyanotic appearance indicative of respiratory problems.
Mutant lungs dissected from 18.5 dpc embryos, were normally developed with respect to number of lobes. Cross-sectioning through the lung revealed that the major bronchial trees had formed in the homozygotes, but they were more compact and smaller compared with wild type (Fig. 8A,B). At higher magnification, it became apparent that the prealveolar sacculi were immature and only poorly inflated, and mesenchymal thickening between the terminal airspaces was observed (Fig. 8C,D). Interaction between mesenchyme and epithelium is a crucial factor throughout lung development. Yet, between 11.5 and 13.5 dpc, no differences in number of buds or sizes of the lungs were observed compared with control embryos (Fig. 8E,F). The presence of surfactant protein C (SP-C) positive cells in 18.5 dpc mutant lungs further indicated that differentiation of precursor epithelial airway cells had proceeded (Fig. 8G,H).
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DISCUSSION |
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The poor retention of nidogen 1 in the majority of basement membrane structures of 1III4-deficient tissues was not expected, although the protein level determined was normal. Nidogen 1 has a highly versatile binding repertoire for other basement membrane components (Timpl and Brown, 1996
), including a high-affinity interaction to the perlecan core protein (Hopf et al., 1999
). The finding that normal immunoreactivity was only apparent in a few basement membrane structures, strongly argues that nidogen 1 is preferably integrated into basement membranes via laminins containing the
1 chain.
It has previously been shown that inactivation of the Nid1 gene in mice and C. elegans did not interfere with basement membrane formation and the animals are viable and were fertile (Murshed et al., 2000; Kang and Kramer, 2000
). Yet, compensation by nidogen 2, which has a highly homologous domain structure and binding repertoire (Kohfeldt et al., 1998
), might be the reason for the lack of phenotype in Nid1 mutant mice (Murshed et al., 2000
), and, vice versa, the same may be true for mice with a mutation in Nid2 (Mitchell et al., 2001
). Although human nidogen 2 binds with only low affinity to the nidogen 1-binding site within the laminin
1 chain (Kohfeldt et al., 1998
), the murine homolog has now been shown to be similar in binding affinity as nidogen 1 (T. Sasaki and R. Timpl, personal communication). It will therefore be of interest to see whether the double mutation of both genes reflects the phenotype described here for the deletion of the nidogen-binding site. Together, the genetic analyses supports the conclusion that neither nidogen 1 (Murshed et al., 2000
; Kang and Kramer, 2000
) nor, as shown here, its binding site on the laminin
1 chain are crucial for basement membrane assembly and function in most tissues. Only a few tissues, including the cortex and neural tube, showed subtle discontinuities in basement membranes and developmental abnormalities (Halfter et al., 2002
), arguing for a specific physiological function of the laminin-nidogen interaction. However, we cannot exclude the possibility that other, presently unknown functions of the nidogen-binding module of the laminin
1 chain might contribute to the described phenotype.
The laminin-nidogen interaction is crucially important for Wolffian duct growth
Genetic data obtained from human diseases and transgenic mice have implicated transcription and growth factors, cell-surface receptors and extracellular matrix components in metanephric development (Lechner and Dressler, 1997; Müller and Brändli, 1999
). Renal agenesis to a variable degree was manifested by mutations in several of these genes. Perturbed communication between cellular receptors and their respective ligands on either the metanephric blastema or the ureteric bud have been suggested in null-mutations for the glial cell line-derived neurotrophic factor, GDNF, and the tyrosine kinase receptor Ret (Lechner and Dressler, 1997
), as well as in integrin
8-deficient mice (Müller et al., 1997
), leading to defective metanephric kidney induction. However, renal agenesis in the absence of the laminin-nidogen 1 interaction is due to defects in Wolffian duct elongation and therefore manifests earlier during development.
The Wolffian duct develops from the intermediate mesoderm and gives rise to parts of the male genital system. In agreement with a primary defect in Wolffian duct growth, vas deferens and the seminal vesicles in male mutant embryos were missing, while the testes were normally present. The female genital tracts originate from the Müllerian duct, which forms later in development in parallel to the Wolffian duct, and its growth was shown to be dependent on an inductive influence of the former (Jacob et al., 1999). In support of these data, we found both ducts to be blind-ending in the urogenital ridge in 13.5 dpc embryos and the female mutants to lack the uterus and occasionally the oviduct.
The Wolffian duct develops in the absence of the laminin-nidogen interaction, excluding its contribution to the first mesenchymal-to-epithelial transition that occurs in kidney development (Saxen, 1987), but its growth caudally towards the cloaca is inhibited. A similar phenotype was described for the targeted ablation of Pax2, a member of the paired-box family of regulatory transcription factors. As
1III4-deficient mice, Pax2 mutants fail to form parts of the genital tract and the ureteric bud (Torres et al., 1995
), suggesting that growth of the Wolffian duct is affected in a similar manner. One possibility therefore is that deletion of the nidogen-binding site interferes with Pax2 expression in the epithelial tube. Yet, normal immunostaining of Pax2 in the mutant Wolffian duct indicates that it is not affected through the deletion.
Although we do not know the detailed mechanism responsible for the phenotype of our 1III4 mutants, these mice now provide a model system to further our understanding of Wolffian duct growth and elongation, which has long been a subject of interest because of its central position in urogenital development. Once formed, the duct progressively elongates; so far two mechanisms have been proposed for this process: one suggests autonomous growth through cell proliferation at the tip of the duct, whereas a second model proposes that mesenchymal cells are continuously added by an epithelial transition (Saxen, 1987
). The data presented here provide evidence that Wolffian duct elongation through autonomous cell proliferation at its growing tip can be excluded. First of all, the number of BrdU-positive cells were similar in mice with the nidogen-binding site deletion as in controls, and did not accumulate at the tip of the growing duct, but uniformly spread over the length of the duct as it has been described in amphibians (Overton, 1959
), indicating that the underlying mechanisms are not due to reduced cell proliferation. Strikingly, however, we identified subtle local ruptures in the basement membrane close to the tip of the duct that may well interfere with duct elongation. Nidogen-binding to laminin could modulate the conformation of laminin or affect the spatial relationships between basement membrane components, which facilitate their interaction with cellular receptors. We therefore propose that the locally restricted discontinuities seen in basement membranes around the Wolffian duct are due to a perturbed cell-matrix interaction, which in turn interferes with signaling cascades necessary to induce the genetic program to recruit mesenchymal cells.
The function of the laminin-nidogen interaction for epithelial branching morphogenesis in kidney and lung
The laminin-nidogen interaction has been suggested to be crucial for epithelial branching morphogenesis in general (Ekblom et al., 1994; Kadoya et al., 1997
). Our data indicate that early branching morphogenesis of the lung is not disturbed in the absence of the laminin-nidogen interaction. However, shortly before birth, the lungs were only poorly inflated, with a mesenchymal thickening of the distal airspaces. Consequently, a respiratory problem caused with high probability the death of all the mutant pups. Interestingly, a similar phenotype has been reported in the absence of TGFß3 (Kaartinen et al., 1995
). The phenotype in TGFß3-deficient mice has been correlated with reduced number of airway precursor cells (Shi et al., 1999
). Although we identified these cells in
1III4-deficient lungs by staining for SP-C, a detailed statistical analysis at different developmental stages is required to determine whether they are formed in similar numbers as in wild type. Ultrastructural analysis demonstrated that the majority of the distal airspaces in the mutants were uninflated, disorganized structures containing epithelial and mesenchymal cells. Strikingly, those sacculi that had formed the alveolar basement membranes showed severe abnormalities, being either amorphously deposited or absent. There are several potential explanations for this finding. A perturbed laminin-nidogen interaction could result in a differentiation deficit of airway precursor cells into type II and type I alveolar pneumocytes, or, alternatively, might interfere with their polarization. Cell contact with basement membranes has been suggested to be regulatory for sorting proteins to the apical or basolateral surfaces (Rodriguez-Boulan and Nelson, 1989
). A possible scenario could therefore be that in the mutants, cell-surface receptors are wrongly translocated to the apical surface, and subsequently basement membrane assembly and formation of the air-blood barrier cannot take place. Further studies with in vitro lung organ cultures will be needed to distinguish between these possibilities.
Stochastic events are most likely to be behind observations that a proportion of mice with inactivated genes can to some extent overcome developmental problems. For example, deficiencies for the integrin 4 and
v subunits or perlecan, have independent lethal phenotypes at different developmental stages, depending on the individual animal (Yang et al., 1995
; Bader et al., 1998
; Costell et al., 1999
). Furthermore, variable penetrance of impaired kidney development has been observed upon mutations of the integrin
8 subunit, Ret or GDNF (Schuchardt et al., 1996
; Moore et al., 1996
; Müller et al., 1997
). Similarly, a minority of the
1III4-deficient animals displayed metanephric development, which may be due to either intact or regenerated basement membranes along the Wolffian duct at crucial stages of development. However, the number of tubuli and glomeruli in the mature part of the cortex in 18.5 dpc mutant kidneys was reduced even in those mutants in which induction of the metanephric blastema and branching of the ureteric bud proceeded without any obvious defects. Inhibition of the laminin-nidogen interaction by antibodies led to reduced tubulogenesis in an in vitro organ culture model, although in contrast to the present observations in vivo, basement membranes were largely distorted in the in vitro analysis (Ekblom et al., 1994
). Thus, although the basement membrane architecture in the
1III4-deficient kidneys appeared normal, it is tempting to speculate that locally restricted and transient basement membrane ruptures also occur during kidney morphogenesis, resulting in the differentiation defect.
Unexpectedly, the bladder of the mutant animals was empty even when kidneys had developed. The histological analysis in male mice demonstrated that the ureter failed to open into the bladder but instead stayed connected aberrantly either to the vas deferens or the epididymis, both derivatives of the Wolffian duct. The cysts seen in these kidneys are therefore likely to be secondary to a defect in urine outflow. These results also indicate that the reduction of the glomerular tufts is attributed to increased pressure through a tailback of the urine, rather than by a primary defect in mesangial cells as described in Pdgfb mutant mice (Leveen et al., 1994), and their GBM may rupture because of mechanical stress. Interestingly, this phenotype resembles congenital anomalies of the kidney and urinary tract (CAKUT) found in humans and mice (Pope et al., 1999
). An abnormal ureter connection in conjunction with a hydronephric kidney has been reported for heterozygous Bmp4 mutant mice (Miyazaki et al., 2000
). In these mice it is suggested that a deficit in BMP4 levels inhibits branching of the ureterovesicle junction into the cloaca as a result of impaired elongation of the ureter (Miyazaki et al., 2000
). This model is reminiscent to the growth defect of the Wolffian duct in the urogenital ridge in the
1III4 mutant animals. We therefore propose that a similar mechanism to that described above leads to the aberrant ureter fusion. However, we cannot exclude at the moment that the absence of the laminin-nidogen interaction causes subtle changes in the supramolecular organization of basement membranes, which then in turn could interfere with the sequestration of BMP4 or other growth and morphogenetic factors, or that specific unknown physiological functions exist for the nidogen-binding module,
1III4 of the laminin
1 chain
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ACKNOWLEDGMENTS |
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REFERENCES |
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Bader, B. L., Rayburn, H., Crowley, D. and Hynes, R. O. (1998). Extensive vasculogenesis, angiogenesis, and organogenesis precede lethality in mice lacking all v integrins. Cell 95, 507-519.[Medline]
Baumgartner, R., Czisch, M., Mayer, U., Pöschl, E., Huber, R., Timpl, R. and Holak, T. A. (1996). Structure of the nidogen binding LE module of the laminin 1 chain in solution. J. Mol. Biol. 257, 658-668.[Medline]
Costell, M., Gustafsson, E., Aszodi, A., Mörgelin, M., Bloch, W., Hunziker, E., Addicks, K., Timpl, R. and Fässler, R. (1999). Perlecan maintains the integrity of cartilage and some basement membranes. J. Cell Biol. 147, 1109-1122.
DiPersio, C. M., Hodivala-Dilke, K. M., Jaenisch, R., Kreidberg, J. A. and Hynes, R. O. (1997). 3ß1 integrin is required for normal development of the epidermal basement membrane. J. Cell Biol. 137, 729-742.
Dziadek, M. and Timpl, R. (1985). Expression of nidogen and laminin in basement membranes during mouse embryogenesis and in teratocarcinoma cells. Dev. Biol. 111, 372-382.[Medline]
Ekblom, P., Ekblom, M., Fecker, L., Klein, G., Zhang, H. Y., Kadoya, Y., Chu, M. L., Mayer, U. and Timpl, R. (1994). Role of mesenchymal nidogen for epithelial morphogenesis in vitro. Development 120, 2003-2014.
Fox, J. W., Mayer, U., Nischt, R., Aumailley, M., Reinhardt, D., Wiedemann, H., Mann, K., Timpl, R., Krieg, T., Engel, J. and Timpl, R. (1991). Recombinant nidogen consists of three globular domains and mediates binding of laminin to collagen type IV. EMBO J. 10, 3137-3146.[Abstract]
Halfter, W., Dong, S., Yip, L., Willem, M. and Mayer, U. (2002). A critical function of the pial basement membrane in cortical histogenesis. J. Neurosci. (in press).
Helbling-Leclerc, A., Zhang, X., Topaloglu, H., Cruaud, C., Tesson, F., Weissenbach, J., Tome, F. M., Schwartz, K., Fardeau, M., Tryggvason, K. and Guicheney, P. (1995). Mutations in the laminin 2-chain gene (LAMA2) cause merosin-deficient congenital muscular dystrophy. Nat. Genet. 11, 216-218.[Medline]
Henry, M. D. and Campbell, K. P. (1998). A role for dystroglycan in basement membrane assembly. Cell 95, 859-870.[Medline]
Hopf, M., Göhring, W., Kohfeldt, E., Yamada, Y. and Timpl, R. (1999). Recombinant domain IV of perlecan binds to nidogens, laminin-nidogen complex, fibronectin, fibulin-2 and heparin. Eur. J. Biochem. 259, 917-925.
Jacob, M., Konrad, K. and Jacob, H. J. (1999). Early development of the müllerian duct in avian embryos with reference to the human. An ultrastructural and immunohistochemical study. Cells Tiss. Organs 164, 63-81.
Kaartinen, V., Voncken, J. W., Shuler, C., Warburton, D., Bu, D., Heisterkamp, N. and Groffen, J. (1995). Abnormal lung development and cleft palate in mice lacking TGF-ß3 indicates defects of epithelial-mesenchymal interaction. Nat. Genet. 11, 415-421.[Medline]
Kadoya, Y., Salmivirta, K., Talts, J. F., Kadoya, K., Mayer, U., Timpl, R. and Ekblom, P. (1997). Importance of nidogen binding to laminin 1 for branching epithelial morphogenesis of the submandibular gland. Development 124, 683-691.
Kallunki, T., Ikonen, J., Chow, L. T., Kallunki, P. and Tryggvason, K. (1991). Structure of the human laminin B2 chain gene reveals extensive divergence from the laminin B1 chain gene. J. Biol. Chem. 266, 221-228.
Kang, S. H. and Kramer, J. M. (2000). Nidogen is nonessential and not required for normal type IV collagen localization in Caenorhabditis elegans. Mol. Biol. Cell 11, 3911-3923.
Koch, M., Olson, P. F., Albus, A., Jin, W., Hunter, D. D., Brunken, W. J., Burgeson, R. E. and Champliaud, M. F. (1999). Characterization and expression of the laminin 3 chain: a novel, non-basement membrane-associated, laminin chain. J. Cell Biol. 145, 605-618.
Kohfeldt, E., Sasaki, T., Göhring, W. and Timpl, R. (1998). Nidogen-2: a new basement membrane protein with diverse binding properties. J. Mol. Biol. 282, 99-109.[Medline]
Kreidberg, J. A., Donovan, M. J., Goldstein, S. L., Rennke, H., Shepherd, K., Jones, R. C. and Jaenisch, R. (1996). 3ß1 integrin has a crucial role in kidney and lung organogenesis. 122, 3537-3547.
Lechner, M. S. and Dressler, G. R. (1997). The molecular basis of embryonic kidney development. Mech. Dev. 62, 105-120.[Medline]
Leveen, P., Pekny, M., Gebre-Medhin, S., Swolin, B., Larsson, E. and Betsholtz, C. (1994). Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes Dev. 8, 1875-1887.[Abstract]
Libby, R. T., Champliaud, M. F., Claudepierre, T., Xu, Y., Gibbons, E. P., Koch, M., Burgeson, R. E., Hunter, D. D. and Brunken, W. J. (2000). Laminin expression in adult and developing retinae: evidence of two novel CNS laminins. J. Neurosci. 20, 6517-6528.
Mayer, U. and Timpl, R. (1994). Structure and function of basement membrane protein nidogen. In Extracellular Matrix Assembly and Structure (ed. P. D. Yurchenco, D. E. Birk and R. E. Mecham), pp. 383-416. San Diego, CA: Academic Press.
Mayer, U., Mann, K., Timpl, R. and Murphy, G. (1993a). Sites of nidogen cleavage by proteases involved in tissue homeostasis and remodelling. Eur. J. Biochem. 217, 877-884.[Abstract]
Mayer, U., Nischt, R., Pöschl, E., Mann, K., Fukuda, K., Gerl, M., Yamada, Y. and Timpl, R. (1993b). A single EGF-like motif of laminin is responsible for high affinity nidogen binding. EMBO J. 12, 1879-1885.[Abstract]
Mayer, U., Pöschl, E., Gerecke, D. R., Wagman, D. W., Burgeson, R. E. and Timpl, R. (1995). Low nidogen affinity of laminin-5 can be attributed to two serine residues in EGF-like motif 2III4. FEBS Lett. 365, 129-132.[Medline]
Mayer, U., Saher, G., Fässler, R., Bornemann, A., Echtermeyer, F., von der Mark, H., Miosge, N., Pöschl, E. and von der Mark, K. (1997). Absence of integrin 7 causes a novel form of muscular dystrophy. Nat. Genet. 17, 318-323.[Medline]
Mayer, U., Kohfeldt, E. and Timpl, R. (1998). Structural and genetic analysis of laminin-nidogen interaction. Ann. New York Acad. Sci. 857, 130-142.
Miner, J. H., Cunningham, J. and Sanes, J. R. (1998). Roles for laminin in embryogenesis: exencephaly, syndactyly, and placentopathy in mice lacking the laminin 5 chain. J. Cell Biol. 143, 1713-1723.
Miner, J. H. and Patton, B. L. (1999). Laminin-11. Int. J. Biochem. Cell Biol. 31, 811-816.[Medline]
Mitchell, K. J., Pinson, K. I., Kelly, O. G., Brennan, J., Zupicich, J., Scherz, P., Leighton, P. A., Goodrich, L. V., Lu, X., Avery, B. J. et al. (2001). Functional analysis of secreted and transmembrane proteins critical to mouse development. Nat. Genet. 28, 241-249.[Medline]
Miyazaki, Y., Oshima, K., Fogo, A., Hogan, B. L. and Ichikawa, I. (2000). Bone morphogenetic protein 4 regulates the budding site and elongation of the mouse ureter. J. Clin. Invest. 105, 863-873.
Moore, M. W., Klein, R. D., Farinas, I., Sauer, H., Armanini, M., Phillips, H., Reichardt, L. F., Ryan, A. M., Carver-Moore, K. and Rosenthal, A. (1996). Renal and neuronal abnormalities in mice lacking GDNF. Nature 382, 76-79.[Medline]
Müller, U. and Brändli, A. W. (1999). Cell adhesion molecules and extracellular-matrix constituents in kidney development and disease. J. Cell Sci. 112, 3855-3867.
Müller, U., Wang, D., Denda, S., Meneses, J. J., Pedersen, R. A. and Reichardt, L. F. (1997). Integrin 8ß1 is critically important for epithelial-mesenchymal interactions during kidney morphogenesis. Cell 88, 603-613.[Medline]
Murray, P. and Edgar, D. (2000). Regulation of programmed cell death by basement membranes in embryonic development. J. Cell Biol. 150, 1215-1221.
Murshed, M., Smyth, N., Miosge, N., Karolat, J., Krieg, T., Paulsson, M. and Nischt, R. (2000). The absence of nidogen 1 does not affect murine basement membrane formation. Mol. Cell. Biol. 20, 7007-7012.
Overton, J. (1959). Studies on the mode of outgrowth of the amphibian pronephric duct. J. Embryol. Exp. Morphol. 7, 86-93.
Paulsson, M., Aumailley, M., Deutzmann, R., Timpl, R., Beck, K. and Engel, J. (1987). Laminin-nidogen complex. Extraction with chelating agents and structural characterization. Eur. J. Biochem. 166, 11-19.[Abstract]
Pope, J. C., Brock, J. W., III, Adams, M. C., Stephens, F. D. and Ichikawa, I. (1999). How they begin and how they end: classic and new theories for the development and deterioration of congenital anomalies of the kidney and urinary tract, CAKUT. J. Am. Soc. Nephrol. 10, 2018-2028.
Pöschl, E., Fox, J. W., Block, D., Mayer, U. and Timpl, R. (1994). Two non-contiguous regions contribute to nidogen binding to a single EGF-like motif of the laminin 1 chain. EMBO J. 13, 3741-3747.[Abstract]
Pöschl, E., Mayer, U., Stetefeld, J., Baumgartner, R., Holak, T. A., Huber, R. and Timpl, R. (1996). Site-directed mutagenesis and structural interpretation of the nidogen binding site of the laminin 1 chain. EMBO J. 15, 5154-5159.[Abstract]
Pulkkinen, L. and Uitto, J. (1999). Mutation analysis and molecular genetics of epidermolysis bullosa. Matrix Biol. 18, 29-42.[Medline]
Ries, A., Göhring, W., Fox, J. W., Timpl, R. and Sasaki, T. (2001). Recombinant domains of mouse nidogen-1 and their binding to basement membrane proteins and monoclonal antibodies. Eur. J. Biochem. 268, 5119-5128.
Rodriguez-Boulan, E. and Nelson, W. J. (1989). Morphogenesis of the polarized epithelial cell phenotype. Science 245, 718-725.[Medline]
Sasaki, T., Wiedemann, H., Matzner, M., Chu, M. L. and Timpl, R. (1996). Expression of fibulin-2 by fibroblasts and deposition with fibronectin into a fibrillar matrix. J. Cell Sci. 109, 2895-2904.
Sasaki, T., Forsberg, E., Bloch, W., Addicks, K., Fässler, R. and Timpl, R. (1998). Deficiency of ß1 integrins in teratoma interferes with basement membrane assembly and laminin-1 expression. Exp. Cell Res. 238, 70-81.[Medline]
Saxen, L. (1987). Organogenesis of the Kidney. Cambridge, UK: Cambridge University Press.
Shi, W., Heisterkamp, N., Groffen, J., Zhao, J., Warburton, D. and Kaartinen, V. (1999). TGF-ß3-null mutation does not abrogate fetal lung maturation in vivo by glucocorticoids. Am. J. Physiol. 277, L1205-L1213.
Schuchardt, A., DAgati, V., Pachnis, V. and Costantini, F. (1996). Renal agenesis and hypodysplasia in ret-k- mutant mice result from defects in ureteric bud development. Development 122, 1919-1929.
Smyth, N., Vatansever, H. S., Murray, P., Meyer, M., Frie, C., Paulsson, M. and Edgar, D. (1999). Absence of basement membranes after targeting the LAMC1 gene results in embryonic lethality due to failure of endoderm differentiation. J. Cell Biol. 144, 151-160.
Stetefeld, J., Mayer, U., Timpl, R. and Huber, R. (1996). Crystal structure of three consecutive laminin-type epidermal growth factor-like (LE) modules of laminin 1 chain harboring the nidogen binding site. J. Mol. Biol. 257, 644-657.[Medline]
Thomas, T. and Dziadek, M. (1993). Genes coding for basement membrane glycoproteins laminin, nidogen, and collagen IV are differentially expressed in the nervous system and by epithelial, endothelial, and mesenchymal cells of the mouse embryo. Exp. Cell Res. 208, 54-67.[Medline]
Timpl, R. and Brown, J. C. (1996). Supramolecular assembly of basement membranes. BioEssays 18, 123-132.[Medline]
Torres, M., Gomez-Pardo, E., Dressler, G. R. and Gruss, P. (1995). Pax-2 controls multiple steps of urogenital development. Development 121, 4057-4065.
Vorbroker, D. K., Profitt, S. A., Nogee, L. M. and Whitsett, J. A. (1995). Aberrant processing of surfactant protein C in hereditary SP-B deficiency. Am. J. Physiol 268, L647-L656.
Williamson, R. A., Henry, M. D., Daniels, K. J., Hrstka, R. F., Lee, J. C., Sunada, Y., Ibraghimov-Beskrovnaya, O. and Campbell, K. P. (1997). Dystroglycan is essential for early embryonic development: disruption of Reicherts membrane in Dag1-null mice. Hum. Mol. Genet. 6, 831-841.
Wobus, A. M., Wallukat, G. and Hescheler, J. (1991). Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers. Differentiation 48, 173-182.[Medline]
Yang, J. T., Rayburn, H. and Hynes, R. O. (1995). Cell adhesion events mediated by alpha 4 integrins are essential in placental and cardiac development. Development 121, 549-560.