Primary hippocampal neurons (DIV 14) were transfected using the Magnetofectamine transfection kit.
This paper demonstrated the high efficiency of Magnetofectamine (association of Lipofectamine 2000 and CombiMag) from
OZ Biosciences to transfect mouse primary hippocampal neurons.
paper reference: Am J Hum Genet. 2013 Apr 24. pii: S0002-9297(13)00129-8.
ZC4H2 Mutations Are Associated with Arthrogryposis Multiplex Congenita and Intellectual Disability through Impairment of Central and Peripheral Synaptic Plasticity.
Hirata H,
Nanda I,
van Riesen A,
McMichael G,
Hu H,
Hambrock M,
Papon MA,
Fischer U,
Marouillat S,
Ding C,
Alirol S,
Bienek M,
Preisler-Adams S,
Grimme A,
Seelow D,
Webster R,
Haan E,
Maclennan A,
Stenzel W,
Yap TY,
Gardner A,
Nguyen LS,
Shaw M,
Lebrun N,
Haas SA,
Kress W,
Haaf T,
Schellenberger E,
Chelly J,
Viot G,
Shaffer LG,
Rosenfeld JA,
Kramer N,
Falk R,
El-Khechen D,
Escobar LF,
Hennekam R,
Wieacker P,
Hübner C,
Ropers HH,
Gecz J,
Schuelke M,
Laumonnier F,
Kalscheuer VM.
Abstract
Arthrogryposis multiplex congenita (AMC) is caused by heterogeneous pathologies leading to multiple antenatal joint contractures through fetal akinesia. Understanding the pathophysiology of this disorder is important for clinical care of the affected individuals and genetic counseling of the families. We thus aimed to establish the genetic basis of an AMC subtype that is
associated with multiple dysmorphic features and
intellectual disability
(ID). We used haplotype analysis, next-generation sequencing, array
comparative genomic hybridization, and chromosome breakpoint mapping to
identify the pathogenic
mutations
in families and simplex cases. Suspected disease variants were verified
by cosegregation analysis. We identified disease-causing
mutations in the zinc-finger gene
ZC4H2
in four families affected by X-linked AMC plus ID and one family
affected by cerebral palsy. Several heterozygous females were also
affected, but to a lesser degree. Furthermore, we found two
ZC4H2
deletions and one rearrangement in two female and one male unrelated
simplex cases, respectively. In mouse primary hippocampal neurons,
transiently produced
ZC4H2
localized to the postsynaptic compartment of excitatory synapses, and
the altered protein influenced dendritic spine density. In zebrafish,
antisense-morpholino-mediated
zc4h2 knockdown caused abnormal swimming and impaired α-motoneuron development. All missense
mutations identified herein failed to rescue the swimming defect of zebrafish morphants. We conclude that
ZC4H2 point
mutations, rearrangements, and small deletions cause a clinically variable broad-spectrum neurodevelopmental disorder of the
central and
peripheral nervous systems in both familial and simplex cases of both sexes. Our results highlight the importance of
ZC4H2 for genetic testing of individuals presenting with ID plus muscle weakness and minor or major forms of AMC.
The Magnetofectamine kit from OZ Biosciences Ideal to transfect primary and Hard-to-transfect cells. Magnetofectamine™ associates Lipofectamine™ 2000 from Invitrogen™ and OZ Biosciences's CombiMag reagent, increasing the overall efficiency of your transfection while minimizing cytotoxicity.
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