Transgenic mice expressing mutated full-length HD cDNA: a paradigm for locomotor changes and selective neuronal loss in Huntington's disease

Huntington's disease (HD) is a progressive neurodegenerative disorder chara cterized clinically by motor and psychiatric disturbances and pathologicall y by neuronal loss and gliosis (reactive astrocytosis) particularly in the striatum and cerebral cortex. We have recently created HD full-length cDNA transgenic mouse models that may serve as a paradigm for HD. A more detaile d characterization of these models is presented here. The transgene encodin g nor mal huntingtin consists of 9417 bp of the huntingtin coding sequences including 16 tandem CAGs coding for polyglutamines as part of exon 1. The transgene is driven by a heterologous cytomegalovirus promoter. Five indepe ndent transgenic mouse lines were obtained using this construct. An additio nal six transgenic lines were obtained using full-length HD constructs that have been modified to include either 48 or 89 CAG repeat expansions. South ern blot and densitometric analyses indicated unique integration sites for the transgene in each of the lines with a copy number ranging from two to 2 2 copies. Widespread expression of the transgene in brain, heart, spleen, k idney, lung, liver and gonads from each line was determined by Western blot analyses. In the brain, transgene expression was found in cerebral cortex, striatum, hippocampus and cerebellum. Expression of the transgene was as m uch as five times the endogenous mouse huntingtin level. Phenotypically, only mice expressing 48 or 89 CAG repeats manifested progre ssive behavioural and motor dysfunction. Early behavioural abnormalities we re characterized by trunk curling and clasping of both fore- and hindlimbs when the animals were suspended by their tails. Subsequently, these mice ex hibited hyperkinetic movements, including heightened exploratory activities , unidirectional rotational behaviour, backflipping and excessive grooming that lasted for several weeks. Eventually, the animals progressed to a hypo kinetic phase consisting of slowed movements and lack of response to sensor y stimuli. Urine retention or incontinence was also a prominent feature of the hypokinetic phase. At the end stage of the disease process, HD48(B,D) a nd HD89(A-C) mice became akinetic just prior to death. Neuropathological examination of mice at various stages indicated that it w as only during the hypokinetic phase and thereafter when selective neuronal loss was most apparent. Regions of neurodegeneration and loss included the striatum, cerebral cortex, thalamus and hippocampus. TUNEL staining indica ted an apoptotic mode of cell death in these brain regions. Comparative neu ronal counts after Nissl staining showed as much as 20% loss of small and m edium neurons in the striatum in mice at the hypokinetic and akinetic stage s. Reactive astrocytosis accompanied the areas of neurodegeneration and los s. Polyglutamine aggregates in the form of neuronal intranuclear inclusions and diffuse nuclear and perinuclear aggregations were found in a small per centage of neurons, including those in brain regions that are typically spa red in HD. This observation suggests that polyglutamine aggregates may not be sufficient to cause neuronal loss in HD. In both behavioural and neuropa thological analyses, wild-type and transgenic animals with 16 CAG repeats w ere indistinguishable from each other and do not exhibit the changes observ ed for mice carrying the 48 and 89 CAG repeat mutations. Thus, animals expr essing the CAG repeat expansions appear to represent clinically analogous m odels for HD pathogenesis, and may also provide insights into the underlyin g pathophysiological mechanisms of other triplet repeat disorders.