Neurobiology of Aging 33 (2012) 2233.e7–2233.

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Negative results

UBQLN2 in familial amyotrophic lateral sclerosis in the Netherlands

Perry T. C. van Doormaal a, Wouter van Rheenen a, Marka van Blitterswijk a,
Raymond D. Schellevis a, Helenius J. Schelhaas b, Marianne de Visser c,
Anneke J. van der Kooi c, Jan H. Veldink a,1, Leonard H. van den Berg a,1,*
a
Department of Neurology, Rudolf Magnus Institute of Neuroscience, University Medical Center Utrecht, the Netherlands
b
Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Nijmegen Medical Center, Nijmegen, the
Netherlands
c
Department of Neurology, Academic Medical Center, Amsterdam, the Netherlands
Received 3 January 2012; received in revised form 1 February 2012; accepted 4 February 2012

Abstract

Recently it was discovered that mutations in the UBQLN2 gene were a cause of an X-linked dominant type of familial amyotrophic lateral
sclerosis (ALS). We investigated the frequency of mutations in this gene in a cohort of 92 families with ALS in the Netherlands. Eight
families were excluded because of male-to-male transmission. In the remaining 84 familial ALS cases no mutations were discovered in
UBQLN2. Hence, UBQLN2 was not found to be a cause of familial ALS in the Netherlands.
© 2012 Elsevier Inc. All rights reserved.

Keywords: Amyotrophic lateral sclerosis; Motor neuron disease; UBQLN2; ubiquilin 2

1. Introduction 2, which functions in the ubiquitinated protein degradation

Amyotrophic lateral sclerosis (ALS) is a neurodegenera-
tive disease with involvement of both upper and lower
motor neurons, resulting in progressive weakness and an
average lifespan of 3 years after disease onset (Mitchell and
Borasio, 2007). In 5%–10% of the cases, ALS is of familial
origin, meaning that at least 1 close relative is also diag-
nosed with the disease (Byrne et al., 2011; Valdmanis and
Rouleau, 2008). A number of genes have been reported to
be causal for familial ALS, from SOD1 being the first gene
discovered in 1993 (Rosen et al., 1993) to more recently a
hexanucleotide repeat expansion in C9ORF72 (Jesus-Her-
nandez et al., 2011; Renton et al., 2011) and mutations in
UBQLN2 (Deng et al., 2011). The latter gene, located on the
X-chromosome, encodes the ubiquitin-like protein ubiquilin
* Corresponding author at: Department of Neurology, G03.228, Univer-
sity Medical Center Utrecht, PO Box 85500, 3508 GA Utrecht, the Neth-
erlands. Tel.: ⫹31 88 7557939; fax: ⫹31 30 2542100.
E-mail address: l.h.vandenberg@umcutrecht.nl (L.H. van den Berg).
1
These authors contributed equally to this work.
pathway. A previous study discovered 5 different mutations
in UBQLN2 in 5 families out of 189 probands from families
of non male-to-male transmitted familial ALS. In familial
ALS, sporadic ALS, and ALS with frontotemporal demen-
tia, ubiquilin pathology was found in spinal and brain tissue.
In France, 1 mutation was found in the UBQLN2 gene in a
cohort of 130 familial ALS patients (Millecamps et al.,
2011). This mutation could not be found in the rest of this
patient’s family, thus probably this is not the cause of the
disease in this family. No other mutation in UBQLN2 was
found in the French cohort.
We analyzed a cohort of familial ALS cases to determine
the frequency of UBQLN2 mutations in the Netherlands.
2. Methods
The study population consists of probands from 92 dif-
ferent ALS families in the Netherlands. In 22 of these
families ALS was associated with frontotemporal dementia.
Genomic DNA was extracted from whole blood using stan-

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2233.e8 P.T.C. van Doormaal et al. / Neurobiology of Aging 33 (2012) 2233.e7–2233.e8
dard procedures. Polymerase chain reaction of the complete
UBQLN2 gene, including 125 base pairs of the 5= untrans-
lated region and 293 base pairs of the 3= untranslated region,
was performed on 100 ng genomic DNA with primers as
reported previously (Deng et al., 2011). BigDye Terminator
3.1 sequencing kit (Applied Biosystems, Foster City, CA,
USA), DNA Analyzer 3730XL, and PolyPhred software
version 6.18 (Washington University, Seattle, WA, USA,
Nickerson et al., 1997) were used for sequencing and data
analysis.
3. Results
Probands of 92 families with ALS were available for
genetic analysis. Because UBQLN2-mediated ALS is an
X-linked dominantly inherited disease, a male-to-male in-
heritance pattern is not possible and therefore 8 families
were excluded. In the remaining 84 families the complete
UBQLN2 gene was sequenced. No mutations were found in
any of these patients.
4. Discussion
UBQLN2 mutations were not found in the 84 ALS fam-
ilies with possible X-linked inheritance that were investi-
gated in the Netherlands. Based on this study the contribu-
tion of mutations in UBQLN2 to the total spectrum of
familial ALS may be small.
In the initial report on UBQLN2, mutations were scarce:
only 5 out of 189 probands (2.6%) without male-to-male
transmission had mutations in this gene (Deng et al., 2011).
Importantly, ubiquilin 2 pathology has been shown in the
spinal cords of ALS cases and in the brains of ALS/demen-
tia cases with or without UBQLN2 mutations, thus indicat-
ing a role for defects in the protein degradation pathway in
ALS.
A recent study in France found a result comparable with
those found in our study: no causative mutations in
UBQLN2 were found in French ALS families (Millecamps
et al., 2011). It has been shown that the frequency of specific
gene mutations can differ significantly between countries as
SOD1 mutations are virtually absent as a cause of ALS in
the Netherlands (2 known families) but, in contrast, account
for 23.5% of familial cases in Scandinavia and in the United
States (Andersen et al., 1997; Cudkowicz et al., 1997). It
seems that the prevalence of UBQLN2 mutations also dif-
fers between countries.
To conclude, UBQLN2 is not found to be a cause of
familial ALS in the Netherlands. Further studies are re-
quired to determine differences between populations.
Disclosure statement
All authors declare no conflicts of interest.
Written informed consent was obtained from all individ-
uals and the study was approved by the local ethical com-
mittee.
Acknowledgements
J.H.V. is funded by the Thierry Latran Foundation. The
research leading to these results has received funding from
the European Community’s Health Seventh Framework
Programme (FP7/2007⫺2013) (grant agreement number
259867).
Appendix: A. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/
j.neurobiolaging.2012.02.032.