Pediatric and Developmental Pathology 10, 328334, 2007

DOI: 10.2350/06-07-0134.1
2007 Society for Pediatric Pathology

Achondrogenesis Type IA (Houston-Harris):

A Still-Unresolved Molecular Phenotype
THOMAS AIGNER,1* TILMAN RAU,2 MANUEL NIEDERHAGEN,1 FRANK ZAUCKE,3
MARKUS SCHMITZ,3 UWE POHLS,4 HELMUT STOSS,5 ANITA RAUCH,6 AND
CHRISTIAN T. THIEL6
1
Institute of Pathology, Liebigstrasse 26, 04103, Leipzig, Germany
2
Department of Pathology, Krankenhausstrasse 810, 91054, Erlangen, Germany
3
Center for Biochemistry, University of Cologne, Joseph-Stelzmann Strasse 52, D-50931, Cologne, Germany
4
Department of Gynecology, University of Erlangen, Universitatsstrasse 2123, 91054, Erlangen, Germany
5
Institute of Pathology, Paderborn, Reumontstrae 28, 33102, Paderborn, FRG
6
Department of Genetics, Schwabachanlage 10, 91054, Erlangen, Germany

Received December 20, 2006; accepted January 17, 2007; published online March 22, 2007.

ABSTRACT Key words: cartilage, chondrogenesis, development,

Achondrogenesis type IA (Houston-Harris) is an extreme-
ly rare lethal chondrodysplasia with a characteristic severe
disarrangement of endochondral ossification. The growth
plate cartilage completely lacks columnar-zone formation
and shows chondrocyte expansion due to intracellular
vacuoles. This article on a new case of achondrogenesis
type IA confirms these findings and demonstrates, on the
ultrastructural level, the retention of fine fibrillar material
within the rough endoplasmic reticulum (rER). Molecular
analysis in the presented case of achondrogenesis type IA
did not reveal mutations in the COL2A1 and SLC26A2
genes, which are known to cause achondrogenesis types
IB and type II. Although the extracellular cartilage matrix
was severely altered, all of the investigated matrix
molecules (collagens, aggrecan, matrilins, cartilage oligo-
meric protein [COMP]) showed a normal distribution
pattern. The only exception was type-X collagen, which
was significantly reduced. Overall, our study suggests a
disturbance in cartilage matrix assembly in the present
case due to the retention of some sort of matrix
component within the rER. Presumably, as a consequence
of this event, processes of chondrocyte maturation and
differentiation and endochondral bone formation are
severely affected in this case of achondrogenesis type IA.
*Corresponding author, e-mail:
Thomas.Aigner@medizin.uni-leipzig.de
skeleton
INTRODUCTION
Chondro-osseous dysplasias represent a heteroge-
nous group of rare and sometimes severe dysonto-
genic lesions affecting the skeleton. Besides
showing clinical symptoms ranging from lethal to
rather normal variants, these lesions also
represent interesting in vivo models of gene defects
in the human.. While the genetic defect for many of
these conditions, including achondrogenesis types
IB and type II, has been characterized, very little is
known about achondrogenesis type IA, which is a
lethal skeletal dysplasia. An autosomal recessive
inheritance was presumed in the reported case but
was not proven [1].
Achondrogenesis type IA (Houston-Harris)
was first described by Houston and Harris in 1972
and later classified as such by Borochowitz and
colleagues [2]. Phenotypically, this condition
results in severe skeletal developmental abnormal-
ities with a major defect in endochondral ossifica-
tion [3,4]. Overall, very few cases and
histopathologic features of this dysplasia have
Figure 1. Macroscopical (a,b) and radiologic features (c,d) of the fetal skeleton showing very poorly ossified skull and lateral
elements of the vertebral column with unossified vertebral bodies; short ribs with cupped ends; short clavicles; hypoplastic
scapulae; very hypoplastic iliac bones; short and wide humeri, femura, tibiae, and fibulae with metaphyseal spikes and
irregularities; and very poorly ossified hands and feet (bars: 1 cm).

been described [35]. Two cases were reported
with ultrastructural characterization [4,5]. Typical
radiologic signs of this condition include an
absence or severely retarded ossification of the
vertebral bodies, horizontally orientated short ribs
with multiple fractures (at least at birth), and
extremely short tubular bones with most marked
changes found in the femur, radius, and ulna. The
characteristic histologic phenotype shows vacuo-
lated and slightly enlarged chondrocytes in the
resting zone and a lack of cell-column formation in
the proliferative cartilage zone.
In this study, we present histomorphologic
data on 1 of the very few cases of achondrogenesis
type IA. To our knowledge, this is the 3rd case to
be analyzed by electron microscopy and the 1st
case to be characterized using molecular tools.
CLINICAL CASE
During an apparently normal pregnancy, ultraso-
nographic examination at 14 weeks of gestation
revealed generalized hydrops fetalis, short and
bowed extremities, a flat facial profile with a
missing nasal bone, and a plexus choroideus cyst.
Due to the severity of the condition, the pregnancy
was terminated. Following fetal abortion, x-ray
examination and an autopsy were performed (Fig.
1ad). These showed a female fetus of the expected
trunk size but with severe shortening of the arms
and legs. The abdomen was distended and the
ACHONDROGENESIS TYPE IA (HOUSTON-HARRIS)
calvarium was missing. The internal organs were
unremarkable. The x-rays showed a very poorly
ossified skull and nonossified vertebral bodies;
short ribs with cupped ends; short clavicles;
hypoplastic scapulae; very hypoplastic iliac bones;
and short, wide humeri, femurs, and tibiae. The
fibulae showed metaphyseal spikes and irregular-
ities, and the hands and feet were very poorly
ossified. Taken together, these findings were
indicative of achondrogenesis. The parents were
nonconsanguineous. The family history was unre-
markable except for an uncomplicated hydroceph-
alus internus in the 1st child.
MATERIALS AND METHODS
Tissue preparation
The growth plate specimens (knee and hip joints)
were fixed in 10% formalin, decalcified in 0.3-M
ethylenediaminetetraacetic acid (pH 7.5), dehy-
drated, cleared in xylene, and embedded in
paraffin. Three- to five-micrometer-thick sections
were cut and stored at room temperature until use.
Histochemical methods
Mucopolysaccharides
The cartilage-typical glycosaminoglycans were
visualized by toluidine blue staining (10 minutes,
0.3% toluidine blue [Merck, Darmstadt, FRG]; pH
3.65, room temperature).
329
Table 1. Primary antibodies and enzymatic pretreatments used for immunohistochemical analyses
Antigen Type Dilution Digestion Source

Aggrecan m 1:1000 H, Pt Dr Perris (Avioli, Italy) (11)

AMACO r 1:500 H Sengle and colleagues (12)
Biglycan r 1:1500 H Dr D. Heinegard (Lund, Sweden)
Collagen I r 1:200 H, Pt Synbio (FRG)
Collagen II m 1:1000 H Calbiochem (Bad Soden, FRG)
Collagen II r 1:50 H, P Novacastra (United Kingdom)
Collagen III r 1:1000 H, P Dr Gunzler (Aventis Pharma, FRG)
Collagen VI (a3) r 1:2000 H, Pt Dr R. Timpl (Munchen, FRG)
Collagen IX r 1:2000 H Budde and colleagues 2005 (13)
Collagen X m 1:50 H, Pt Girkontaite and colleagues (14)
COMP r 1:500 H Hedbom and colleagues 1992 (15)
Matrilin-1 r 1:200 H Hauser and Paulsson 1994 (16)
Matrilin-3 r 1:200 H Klatt and colleagues 2000 (17)
Matrilin-4 r 1:200 H Klatt and colleagues (18)
Mib-1/Ki67 m 1:50 Microwave Dako (Denmark)
S-100 r 1:10000 P Dako (Denmark)
Thrombospondin-1 m 1:1000 H Calbiochem (Bad Soden, FRG)
Vimentin m 1:200 Dako (Denmark)

m indicates mouse monoclonal; r, rabbit polyclonal; H, hyaluronidase (2 mg/mL, phosphate-buffered saline [PBS] pH 5, 60 min at 378C); P, pronase (2 mg/mL,
PBS, pH 7.3, 60 min at 378C); Pt, protease XXIV (0.02 mg/mL, PBS, pH 7.3, 60 min at room temperature).

Collagens
The presence of collagens in the extracellular
matrix was demonstrated by Masson-Goldner
staining. For demonstration of glycoproteins, the
diastaseperiodic acid-Schiff (PAS) reaction was
performed.
Transmission electron microscopy
Conventional transmission electron microscopy
was performed according to Spurr [6] and
modified by Schulz (7). Briefly, small cartilage
pieces were fixed with glutaraldehyde-formol
overnight (1.25% glutaraldehyde, 2% formalin)
and cleared in phosphate-buffer (18.76-mg/mL
NH2PO4, 4.28-mg/mL NaOH, pH 7.5). After
postfixation with 1% OsO4 for 2 hours and
rewashing in the phosphate-buffer, the samples
were dehydrated in an acetone series. Then,
samples were infiltrated with a low-viscosity epoxy
resin-acetone series (50% for 30 minutes, 75%
overnight, 2 times 100% for 2 hours) and
embedded in epoxy resin in standard gelatin
capsules [6,8]. For light microscopic control,
semithin sections (0.51lm) were made and
stained with methylene-blue and azure II [9]. For
electron microscopic analysis, ultrathin sections
(50100 nm) were placed on Formvar-laminated
copper grids and stained with uranyl acetate and
lead citrate.
330 T. AIGNER ET AL.
Immunohistochemistry
Deparaffinized sections were enzymatically pre-
treated and incubated with primary antibodies
(Table 1) overnight at 48C. The antibody labeling
was visualized using alkaline-phosphataselabeled
secondary antibodies and 3-hydroxy-2-naphtylacid
2,4-dimethylanilid as a color substrate [10]. Nuclei
were counterstained with hematoxylin. Alterna-
tively, Alexa 488labeled secondary antibodies
(molecular probes) were used for immunofluores-
cence staining. In these cases, nuclei were stained
with 0.1-lg/mL bisbenzimide (Sigma, Munich,
Germany).
Sequencing of nuclear DNA
Genomic DNA was extracted from a chorionic
villous sample of the proband by a standard salt
precipitation method. Maternal contamination was
excluded by typing of 15 microsatellite markers
from 13 different chromosomes with the Powerplex
16 kit (Promega Corp, Mannheim, Germany). The
54 coding exons of COL2A1 and the 2 coding exons
of SLC26A2 were amplified by polymerase chain
reaction (PCR) using intronic and additional exonic
primers for larger exons. Primers were designed
using the Primer3 software (http://frodo.wi.mit.edu/
cgi-bin/primer3/primer3_www.cgi) based on the
reference sequences COL2A1: NM_001844 and
NT_029419; SLC26A2: NM_000112 and
NT_029289. Primer sequences are available upon
request. The PCR mix consisted of 1 3 PCR-Buffer
(Gibco, Ivitrogen, Karlsruhe, Germany), 800-lM
dNTP-mix (200 lM per nucleotide), 5 pmol of each
primer, 0.12-U Taq-Polymerase (Promega Corp),
and 20-ng template DNA. Reactions were carried
out in an MJ Research thermocycler (MJ Research,
Soeborg, Denmark) using touchdown PCR with a
temperature gradient 658C down to 558C. The PCR
products were purifi ed with Filter Plates
LSKMPCR50 (Millipore, Billerica, MA, USA) on
a Tecan liquid handling system (Tecan, Crailsheim,
Germany). Sequencing reactions were performed
on both strands using the BigDye Terminator Cycle
Sequencing Kit v3.1 (Applied Biosystems, Foster
City, CA) according to the manufacturers instruc-
tions. After purifi cation with fi lter plates
LSKS09624 (Millipore), the products were ana-
lyzed on an ABI Genetic Analyzer 3730 (Applied
Biosystems) with the SeqMan II software (DNAS-
TAR, Madison, WI, USA) and compared with the
reference sequences (COL2A1: NM_001844 and
NT_029419; SLC26A2: NM_000112 and
NT_029289) of The National Center for Biotech-
nology Information (NCBI) database.
RESULTS
Histologically, the fetal growth cartilage was
severely disorganized (Fig. 2a). The columnar zone
was absent, and no clear transition to the hypertro-
phic zone could be delineated. Instead, in the resting
zone, larger chondrocytes were noted containing
inclusion vacuoles. The cartilage matrix appeared
histologically unremarkable. However, very little
bone formation was observed. Some woven bone
was found (Fig. 2b), but no primary spongiosa (i.e.,
with central cartilage cores) was detectable.
Histochemically, the cartilage matrix stained
positive for proteoglycans and collagens, though
this was impossible to evaluate quantitatively. The
cellular vacuole inclusions stained positive in the
diastase-PAS reaction (Fig. 2c).
Ultrastructurally, the extracellular matrix of
the growth plate cartilage showed a reduction of
the collagen content in terms of number and
thickness of the collagen fibers (Fig. 2d). Intracel-
lularly, the chondrocytes showed a dilated rough
endoplasmic reticulum (rER) (Fig. 2eg), most
prominent in the enlarged cells. The rER showed a
fine fibrillar content as well as amorphous material.
No evidence of banding was found. Of note, cells
located in the presumptive hypertrophic zone
tended to show fewer, but larger dilated rER
structures (Fig. 2g). Other cells (e.g., osteocytes,
osteoblasts, hematopoietic and blood cells, endo-
thelial cells, fibroblasts) appeared to be normal and
showed no dilated rER (Fig. 2h).
Immunohistochemically, the extracellular car-
tilage matrix stained normal for all tested matrix
constituents such as aggrecan; collagen type II, VI,
IX and XI; COMP; and others (Table 1) (Fig. 3).
Type-X collagen was found only focally in the very
late calcified cartilage zone (Fig. 3b). The
pericellular matrix of all dilated cells in the resting
zone and most enlarged chondrocytes in the
hypertrophic zone stained negative for type-X
collagen. None of the antibodies (Table 1) showed
a cellular staining pattern in the areas of dilated
chondrocytes. Mib-1 immunohistochemistry
showed a diffuse positive staining throughout the
fetal growth plate cartilage including the ballooned
chondrocytes (also of the presumptive hypertrophic
zone) (Fig. 2i).
Sequencing of the COL2A1 gene revealed 17
va r i a t i o n s : ( I V S 91 5 G / G ( r s 1 0 3 4 7 6 2 ) ,
IVS1115A/G (rs10875716), IVS1683A/A
(rs1793915), IVS16107C/C (rs1635534),
c.1062G/G (p.G354G) rs1793927, c.1314C/C
( p . G 3 6 9 G ) r s 1 7 9 3 9 3 4 , I V S 2 38 7 T / C
(rs1635544), IVS28-46C/C (rs1635550), IVS31-
49G/T (rs11168338), c.2295C/T (G765G)
rs2276454, IVS34-32T/C (rs2276456), IVS34-
22G/A (rs2276457), c.2400T/C (p.N800N)
rs1635553, IVS38-69C/C (rs1635554), c.2673C/
G (p.G891G), c.2805C/T (p.S935S), IVS5342C/
T (rs1635560)). Sequencing of the SLC26A2 gene
showed 1e variation (c.2065A/T (p.T689S)
rs3776070). All variations are known to occur in
nonaffected controls as well and are already
described as polymorphisms in public databases
(Single Nucleotide Polymorphism database at
NCBI; http://www.ncbi.nlm.nih.gov/). The nucleo-
tides at position c.2673 and c.2805, as well as the
amino acid serine at position p.935, are not
conserved throughout the species. Both variations
are rare with a minor allele frequency of below 2%.
DISCUSSION
Our case showed typical radiologic signs of
achondrogenesis while histologic and cytologic
ACHONDROGENESIS TYPE IA (HOUSTON-HARRIS) 331
Figure 2. a. Histologic overview showing enlarged cells throughout the cartilage, including the resting zone (note: no
columnar arrangement of cells is visible). b. Only woven bone without cartilaginous cores is detectable. c. Enlarged
chondrocytes in the resting zone contain period acid-Schiffpositive vesicles. d. Ultrastructural analysis of the extracellular
cartilage matrix showing reduced collagen fibrils of reduced diameters and length. The ground substance appears to be
scarce. eg. Ultrastructural analysis of chondrocytes showing dilated rough endoplasmic reticulum. h. Ultrastructural
analysis shows normal fibroblasts (missing rough endoplasmic reticulum dilatation) surrounded by an unremarkable
collagenous matrix. i. Demonstration of proliferative activity including the hypertrophic zone chondrocytes
(immunostaining for Mib-1/Ki67) (magnification bars: a. 500 lm; b. 100 lm; c,f. 15 lm; d,e. 0.2 lm; g,i. 4 lm; h,l, 5 lm).
A color version of this figure is available in the online journal.

investigations confirmed the characteristic features
of achondrogenesis type IA (Houston-Harris) [3,4].
These features include enlarged, vacuolated chon-
drocytes in the resting zone of the fetal growth
plate cartilage, an overall severe disturbance of
endochondral ossification, and a severe reduction
of bone formation. Disruption of the collagenous
pericellular matrix (as seen in achondrogenesis
type IB) or a significantly reduced cartilage matrix
content (as seen in achondrogenesis type II) was
detectable. The vacuolated chondrocytes showed
histomorphologic signs of intracellular retention of
proteinaceous material [4]. At the ultrastructural
level, a dilation of the rER containing filamentary
and amorphous material was seen. This observa-
332 T. AIGNER ET AL.
tion was only partly in agreement with the case by
Molz and Spycher [4], who found only amorphous
material, and the case described by Yang and
colleagues. [5], who found fibrillar matrix material
identical to that found in extracellular cartilage.
Also, the reduced content of collagens and
proteoglycans within the cartilage matrix on the
ultrastructural level was not observed by Molz and
Spycher [4]. Whether this heterogeneity of cases
reflects different alterations in 1 affected gene or
alterations in different genes leading to a similar
phenotype requires further investigations.
None of the investigated cartilage matrix
components appeared to be missing, and sequenc-
ing of the type II collagen gene did not reveal any
Figure 3. Immunohistochemical analysis of collagen types I (a), II (d), VI (e), IX (f), and X (b) as well as aggrecan (c) , matrilin-
1 (g) and -3 (h) and COMP (i) (magnification bars: a. 100 lm; b,dj. 50 lm; c. 25 lm). A color version of this figure is available
in the online journal.
unusual mutations. However, there was a reduction
in collagen type II fibrils, which also appeared to
be thinner than in normal age-matched fetal
cartilage (own unpublished results). Thus, the
affected gene could encode a proteinnot inves-
tigated in this studywhich is involved in
regulating collagen fiber formation and diameter.
Interestingly, in addition to a severe derangement
of the growth plate structure and morphology, the
maturation of chondrocytes to hypertrophic cells
was impaired as indicated by the delayed expres-
sion of type-X collagen. Also, the proliferative
activity of the chondrocytes was not arranged in a
zonal pattern. As such, positive staining of the
proliferation-associated antigen Ki-67/Mib-1 was
found throughout all areas including the presump-
tive hypertrophic zone. Such alterations in
cellular phenotype and maturation patterns in
combination with deranged matrix formation are
not a surprising finding as many studies have
shown that processes of cell differentiation and
matrix formation are co-regulated. This has been
shown in vitro during dedifferentiation of
chondrocytes [19,20] and in vivo where a mutation
in the collagen type II gene leads to a severe
disturbance of the chondrocyte maturation program
[21] as seen in achondrogenesis type II.
The gene defect of achondrogenesis type IA
is not known so far, whereas those of achondro-
genesis type IB and II have been characterized.
Defects in the collagen type II gene [21,22] lead to
a reduction in collagen type II fibrils within the
extracellular cartilage matrix corresponding to the
typical histologic finding in achondrogenesis type
II. Defects in the SLC26A2e (diastrophic dysplasia
sulphate transporter) gene [23] result in a reduction
ACHONDROGENESIS TYPE IA (HOUSTON-HARRIS) 333
of the proteoglycan content of the cartilage matrix
in achondrogenesis type IB. As a consequence, the
osmotic swelling pressure lowers, and the collagen
network collapses, resulting in formation of typical
pericellular collagenous rings. None of these
phenotypic changes were observed in the present
case of achondrogenesis type I, and neither the
collagen type II or SLC26A2e gene was affected.
However, the affected gene is most likely specific
to chondrocytes, as no cytologically aberrant cells
were found in any other cells/tissue.
Overall, our study suggests a disturbance in
cartilage matrix assembly in achondrogenesis type
IA due to the retention of some sort of matrix
components and/or other molecules within the
rER. Subsequently, disruption of chondrocyte
maturation and matrix synthesis results, followed
by defective endochondral bone formation.
ACKNOWLEDGMENTS
This work was supported by the Deutsche
Forschungsgemeinschaft (grant Ai20/71) and the
Federal Ministry for Education and Research
network grant skeletal dysplasias (SKELENET)
(GFGM01141901 to A. R.). We thank R. Lissner
and F. Boggasch for expert technical assistance and
A. McAlindon for critical reading of the manu-
script.
REFERENCES
1. Superti-Furga A. Achondrogenesis type 1B. J Med Genet 1996;33:
957961.
2. Borochowitz Z, Lachman R, Adomian GE, Spear G, Jones K, Rimoin
DL. Achondrogenesis type I: delineation of further heterogeneity and
identification of two distinct subgroups. J Pediatr 1988;112:2331.
3. Yang SS, Heidelberger KP, Bernstein J. Intracytoplasmic inclusion
bodies in the chondrocytes of type I lethal achondrogenesis. Hum
Pathol 1976;7:667673.
4. Molz G, Spycher MA. Achondrogenesis type I: light and electron-
microscopic studies. Eur J Pediatr 1980;134:6974.
5. Yang SS, Heidelberger KP, Brough AJ, Corbett DP, Bernstein J.
Lethal short-limbed chondrodysplasia in early infancy. Perspect
Pediatr Pathol 1976;3:140.
334 T. AIGNER ET AL.
6. Spurr AR. A low-viscosity epoxy resin embedding medium for
electron microscopy. J Ultrastruct Res 1969;26:3143.
7. Schulz A. A reliable method of preparing undecalcified human bone
biopsies for electron microscopic investigation. Microsc Acta 1977;
30:718.
8. Peabody TD, Gibbs CP, Simon MA. Evaluation and staging of
musculoskeletal neoplasms. J Bone Joint Surg 1998;80-A:1204
1218.
9. Richardson MK, Jarrett JA, Finke E. Embedding in epoxy resins for
ultrathin sectioning in electron microscopy. Stain Technol 1960;35:
313325.
10. Aigner T, Dertinger S, Belke J, Kirchner T. Chondrocytic cell
differentiation in clear cell chondrosarcoma. Hum Pathol 1996;27:
13011305.
11. Aigner T, Loos S, Inwards CY, et al. Chondroblastoma is an osteoid-
forming, but not cartilage-forming neoplasm. J Pathol 1999;189:463
469.
12. Sengle G, Kobbe B, Morgelin M, Paulsson M, Wagener R.
Identification and characterization of AMACO, a new member of
the von Willebrand factor A-like domain protein superfamily with a
regulated expression in the kidney. J Biol Chem 2003;278:50240
50249.
13. Budde B, Blumbach K, Ylostalo J, et al. Altered integration of
matrilin-3 into cartilage extracellular matrix in the absence of
collagen IX. Mol Cell Biol 2005;25:1046510478.
14. Girkontaite I, Frischholz S, Lammi P, et al. Immunolocalization of
type X collagen in normal fetal and adult osteoarthritic cartilage with
monoclonal antibodies. Matrix Biol 1996;15:231238.
15. Hedbom E, Antonsson P, Hjerpe A, et al. Cartilage matrix protein. J
Biol Chem 1992;267:61326136.
16. Hauser N, Paulsson M. Native cartilage matrix protein (CMP): a
compact trimer of subunits assembled via a coiled-coil alpha-helix. J
Biol Chem 1994;269:2574725753.
17. Klatt AR, Nitsche DP, Kobbe B, Morgelin M, Paulsson M, Wagener
R. Molecular structure and tissue distribution of matrilin-3, a
filament-forming extracellular matrix protein expressed during
skeletal development. J Biol Chem 2000;275:39994006.
18. Klatt AR, Nitsche DP, Kobbe B, Macht M, Paulsson M, Wagener R.
Molecular structure, processing, and tissue distribution of matrilin-4.
J Biol Chem 2001;276:1726717275.
19. von der Mark K, Gauss V, von der Mark H, Muller PK. Relationship
between cell shape and type of collagen synthesized as chondrocytes
lose their cartilage phenotype in culture. Nature 1977;267:531532.
20. Benya PD, Padilla SR, Nimni ME. Independent regulation of
collagen types by chondrocytes during the loss of differentiated
function in culture. Cell 1978;15:13131321.
21. Dertinger S, Soder S, Bosch H, Aigner T. Matrix composition of
cartilaginous anlagen in achondrogenesis type II (Langer-Saldino).
Front Biosci 2005;10:446453.
22. Godfrey M, Hollister DW. Type II achondrogenesis-hypochondro-
genesis: identification of abnormal type II collagen. Am J Hum Genet
1988;43:904913.
23. Superti-Furga A, Hastbacka J, Wiilcox WR, et al. Achondrogenesis
type IB is caused by mutations in the diastrophic dysplasia sulphate
transporter gene. Nat Genet 1996;12:100102.