Species: Mouse
Genes: Park2
Modification: Park2: Knock-Out
Disease Relevance: Parkinson's Disease
Strain Name: B6.129S4-Prkn^tm1Shn/J
Genetic Background: C57BL/6J
Availability: Available through The Jackson Laboratory, Stock# 006582, Live and cryopreserved.

Summary

These parkin knockout (KO) mice were generated by creating a germline disruption in the parkin gene within exon 3 leading to a truncated transcript coding for the first 95 amino acids of mouse parkin protein fused in-frame with the coding sequence of EGFP (Goldberg et al., 2003). Loss-of-function mutations in parkin are causally linked to early-onset Parkinson’s disease (Kitada et al., 1998).

Overall Health

Parkin KO mice are viable, fertile, and do not contain any obvious abnormalities (Goldberg et al., 2003). However, their rate of weight gain is delayed, with weight being significantly lower than that of wild-type controls, especially after 6 months of age (Hollville et al., 2020; Palacino et al., 2004). This effect seems to be sex-dependent, with male but not female parkin KO mice exhibiting reduced body weight and fat mass (Moore et al., 2022). The weights of 4-week-old hindlimb muscle tissues were also reduced in male parkin KO mice versus wild-type mice (females were not examined in this study), as was myofiber cross-sectional area but not myofiber number (Peker et al., 2018).

Further reading on Overall Health
(full references listed at the end under Further Reading)

• Peker et al., 2022 (exacerbated fasting-induced skeletal muscle wasting)

Neuropathology

Parkin KO exhibit largely normal brain morphology (Goldberg et al., 2003). At 12, 18, and 24 months of age, the number of tyrosine hydroxylase (TH)-positive neurons did not differ in the substantia nigra (SN) between wild-type and parkin KO mice using unbiased stereology; neuron volumes were also similar between genotypes at 24 months of age. TH staining of the locus coeruleus was generally similar between KO and wild-type mice as well, as was the cell morphology. Dopaminergic projections in the striatum were also normal, based on TH staining.

Inclusions of α-synuclein and ubiquitin were not observed in any brain region (Goldberg et al., 2003).

Spinal cord staining of GFAP (glial fibrillary acidic protein), a marker of astrogliosis, did not differ between non-transgenic and parkin KO mice at 130 days of age (Palomo et al., 2018, also see this paper for outcomes in a double mutant cross with SOD1-G93A mice on mitochondrial function and amyotrophic lateral sclerosis–like disease progression).

Neither the length of serotonergic fibers nor the level of serotonin differed in the hippocampus between 15-month-old parkin KO and wild-type mice (Hennis et al., 2014).

In layer 5 pyramidal neurons of the prelimbic cortex, the complexity of basal dendritic arborization and spine density were reduced in parkin KO mice versus wild-type mice (Huo et al., 2022). However, spine density in hippocampal CA1 pyramidal neurons was not affected. The prelimbic cortex also showed reduced basal neural activity (as measured by c-fos immunostaining) and in response to a sociability task in parkin KO versus wild-type mice at 8 weeks of age.

Further reading on Neuropathology
(full references listed at the end under Further Reading)

• Frank-Cannon et al., 2008 (lipopolysaccharide-induced vulnerability to nigral degeneration and motor function)
• Gunn et al., 2019 (spongiform neurodegeneration in a double mutant cross with Mgrn1 null mice)
• Hwang et al., 2017 (ethanol-induced vulnerability to dopaminergic degeneration and motor function)
• Kitada et al., 2009 (no nigral degeneration in triple knockout parkin/DJ-1/PINK1 mice)
• Liu et al., 2019 (α-synuclein clearance and 20S proteasome activity)
• Pickrell et al., 2015 (neuropathology, motor dysfunction, and mitochondrial dysfunction in a double mutant cross with Mutator mice)
• Pinto et al., 2018 (dopaminergic neurodegeneration, motor defects, and mitochondrial outcomes in a double mutant cross with PD-mito-PstI mice)
• Rogers et al., 2017 (neuromuscular junction degeneration in a double mutant cross with Pink1 KO mice)
• Singh et al., 2018 (astrocyte outcomes following thapsigargin-induced endoplasmic reticulum stress)
• Song et al., 2017 (bioenergetic, neurobehavioral, and dopaminergic outcomes in a double mutant cross with Twinkle-TG mice)

Dopamine Levels

Levels of striatal dopamine and its major metabolites DOPAC (dihydroxyphenylacetic acid) and HVA (homovanillic acid) were similar between parkin KO and wild-type mice at 6, 12, 18, and 24 months of age (Goldberg et al., 2003). However, an increase in striatal extracellular dopamine concentration (as measured by no-net-flux microdialysis) was found in KO mice aged 8 to 9 months. In contrast to these increased basal dopamine levels, evoked dopamine release in 2- to 4-month-old parkin KO mice was decreased compared to wild-type control mice, using amperometric recordings in acute striatal slices (Kitada et al., 2009).

No differences were observed from wild-type mice in striatal dopamine receptor levels, dopamine transporter levels, or binding affinities (Goldberg et al., 2003; Kitada et al., 2009).

Synaptic Function

Striatal medium spiny neurons had reduced synaptic excitability in parkin KO mice at 6-9 months of age, as determined by the higher level of corticostriatal stimulation required to induce a given synaptic response and action potentials (Goldberg et al., 2003). However, other passive and active membrane properties were similar between genotypes.

Intracellular recording of medium spiny neurons revealed that corticostriatal synapses are selectively more sensitive to group II mGlu receptor activation in parkin KO than in wild-type mice, based on evoked excitatory postsynaptic potentials following bath application of the selective mGlu2/3 agonist LY379268 in 8- to 11-week-old animals (Martella et al., 2009).

In another study, long-term depression and long-term potentiation were found to be impaired in 8- to 11-week-old parkin KO mice compared to wild-type control mice, as observed with intracellular recordings of striatal medium spiny neurons (Kitada et al., 2009). However, this was pathway-specific, as long-term potentiation was normal in the hippocampal Schaeffer collateral pathway.

Immunohistochemical analysis of synaptic proteins synaptophysin, Munc-18, and calbindin showed grossly normal staining in parkin KO mice (Goldberg et al., 2003).

The composition of synapses was altered in the brain (Huo et al., 2022). For instance, western blotting revealed that the post-synaptic receptors GluA1, GluA2, and GluN1 were reduced in synaptosome fractions from the frontal cortical area of parkin KO mice compared to wild-type mice, although PSD-95 was not changed. Presynaptically, in layer 5 of the prelimibic cortex, VGLUT1 puncta densities were reduced, meanwhile the puncta density of VGAT was not changed. Other synaptic components were also altered in a region-specific manner (e.g., Kir2.1 was reduced in samples from the frontal cortical area but not of the hippocampal area).

In another study, neurotransmitter receptors were examined using quantitative receptor autoradiography in 12-week-old parkin KO mice versus wild-type mice. Changes were seen in numerous neurotransmitter receptors across 11 brain regions. Most notably, GABAB, kainate receptor, and NMDA receptor densities were increased in multiple areas, while AMPA receptor densities were decreased (Cremer et al., 2015).

Further reading on Synaptic Function
(full references listed at the end under Further Reading)

• Kabayama et al., 2017 (striatal expression of synaptotagmin proteins)

Motor Behavior

General behavior (beam breaks) and exploratory anxiety (stereotypy count), as measured on the open-field test, did not differ between KO and wild-type mice at 6, 12, and 18 months of age (Goldberg et al., 2003). Open-field behavior also did not differ at younger ages based on distance travelled and number of rears (Hollville et al., 2020; also see open-field test results in Huo et al., 2022).

Latency to fall on the accelerating Rotarod, which measures more severe motor dysfunction, did not differ between parkin KO and control mice at 6, 12, or 18 months of age (Goldberg et al., 2003). Interestingly, another study found that 4- to 5-month-old male, but not female, mice performed better on the accelerating Rotarod than wild-type mice (Hollville et al., 2020). Moreover, a different study found that when parkin KO mice were tested at stable, slower speeds, they had a longer latency to fall than wild-type mice, though sex was not specified for this experiment (Hennis et al., 2014).

In contrast to the open-field and Rotarod assays, behavioral paradigms sensitive to nigrostriatal dysfunction, such as the beam traversal task (measured at 2-4, 7, and 18 months of age), revealed deficits in parkin KO mice at all ages compared to wild-type mice (Goldberg et al., 2003). On the adhesive removal test, which assesses somatosensory, in addition to nigrostriatal, function, parkin KO mice also displayed deficits at 2-4 and 7 months of age (but not at 18 months).

Further reading on Motor Behavior
(full references listed at the end under Further Reading)

• Berthet et al., 2012 (abnormal involuntary movements in response to L-DOPA treatment)
• Trabjerg et al., 2023 (motor behavior analysis; also see for non-motor behavior; serum glucose and lipoprotein levels; gene expression analysis of mitochondrial dysfunction, oxidative stress, and inflammation)

Non-Motor Behavior

The Barnes maze test was used to examine hippocampus-dependent spatial memory, and parkin KO mice performed similar to wild-type mice, suggesting comparable learning and memory ability between genotypes (Huo et al., 2022). In a separate study, novel object recognition was significantly decreased in parkin KO mice compared to control mice at 4-5 months of age (Wenqiang et al., 2014).

Sensorimotor function did not differ between parkin KO and wild-type mice based on latency to respond to a hot plate (at 6-7 months of age), latency to find buried food (at 4-5 months of age), and latency to find a visible platform in the Morris water maze (at 4-5 months of age; Hollville et al., 2020). Olfactory function also appears intact in parkin KO mice, based on time spent investigating novel odors in a separate study (Hennis et al., 2014).

Autistic-like behaviors, including reduced sociability, increased repetitive behaviors, and deficits in communication, were present in parkin KO mice at 8 to 12 weeks of age (Huo et al., 2022). Namely, the three-chamber test revealed a reduced preference for social novelty and impairments in social interaction compared to what was exhibited by wild-type mice. On the marble burying test, which measures anxiety-like behaviors, parkin KO mice buried significantly fewer marbles, which may be indicative of reduced anxiety and/or an interest in a new environment. Reduced anxiety-like behavior was also observed on the open-field test, where parkin KO mice spent more time in the inner square area compared to wild-type mice. Stereotyped repetitive self-grooming was observed at a higher frequency and duration in parkin KO mice versus wild-type mice. Abnormalities in ultrasonic vocalization patterns were also observed in parkin KO versus wild-type mice in early developmental periods (postnatal days 5, 7, 9, 11, 13), which was another indication of autistic-like behavior.

Wild-type mice (8- to 13-week-old) exposed to a 30-day chronic social defeat had increased anxiety-like behaviors, but this did not occur in parkin KO mice, as demonstrated through time spent in the center of an open field, the light/dark box, and elevated plus maze (Duan et al., 2021). Social interaction also was not reduced following chronic social defeat stress in parkin KO mice like it was for wild-type mice. Behavioral despair, however, as measured by immobility time on the forced swim test, did increase in both wild-type and KO mice in response to chronic social defeat. In another study, immobility time on the forced swim test also did not differ between 14-month-old parkin KO and wild-type mice (Hennis et al., 2013).

Mitochondrial Abnormalities

Using proteomic analysis of ventral midbrain lysates, a decreased abundance was found in numerous proteins involved in mitochondrial respiration and oxidative stress in parkin KO mice versus wild-type mice at around 8 months of age (Palacino et al., 2004). Functionally, the respiratory capacity (but not efficiency) of striatal mitochondria was reduced in parkin KO mice. However, electron microscopy did not reveal any abnormalities in total number, size, or shape. Furthermore, antioxidant capacity of serum samples from 5- to 12-month-old parkin KO mice was reduced, while reactive oxygen species (ROS) damage in brain samples was increased in 18- to 20-month-old (but not 3-month-old) parkin KO mice compared to wild-type controls.

More recently, the mitochondrial proteome from striatal and liver samples of parkin KO mice was examined in vivo using a longitudinal isotopic metabolic labeling approach (Stauch et al., 2023). This study revealed altered protein expression in synaptic mitochondria from the striatum as well as in mitochondria from the liver from 3-month-old parkin KO mice versus wild-type mice. In synaptic mitochondrial samples, pathway analysis showed that the altered proteins were associated with mechanisms of protein aggregation, repair, and signal transduction. Parkin KO mice also had an overall increase in the abundance and half-life of mitochondrial proteins, though this effect was modest, and the authors concluded that homeostatic regulation of mitochondria was generally not affected by the KO.

Numerous other mitochondrial outcomes have been reported in parkin KO mice (Damiano et al., 2014). For instance, in 24-month-old parkin KO mice, the mitochondrial respiration rate was similar to wild-type mice in samples from the striatum, cortex, and whole brain. However, 24-month-old, but not 9-month-old, parkin KO mice had a decreased respiratory reserve in the striatum, though this reduction was not observed in the midbrain or the liver at either age.

In the amygdala, additional mitochondrial deficits have been observed in parkin KO mice (7- to 24-week-old), including reduced mtDNA copy number, mtDNA transition mutation, and mtDNA replication compared to wild-type mice (Duan et al., 2021).

In dopaminergic neurons of the substantia nigra pars compacta, mitochondrial pathologies (i.e., enlarged mitochondrial inclusions and reduced mitochondria mass) were worsened at 3 weeks of age in parkin KO mice crossed with a mouse that overexpresses human α-synuclein A53T compared to that observed in the α-synuclein A53T–overexpressing mouse with an intact parkin gene (Chen et al., 2015).

In heart tissue, mitochondrial respiration did not differ between parkin KO and wild-type mice at 3 months of age (Kubli et al., 2013). However, ultrastructural analysis of mitochondria in heart tissue (at 12 weeks of age) revealed disorganized mitochondrial networks and smaller mitochondria, though mitochondrial degeneration and damage were not observed.

Skeletal muscle (gastrocnemius) in parkin KO mice displayed decreased levels of oxidative phosphorylation proteins compared to wild-type mice, namely, the complex I protein NDUFB8 and the complex II protein SDHB, as well as decreases in the mRNA levels of certain mitochondrial complex genes (Peker et al., 2018). Moreover, mitochondrial function was impaired in primary myotube cultures from parkin KO mice—KO cells exhibited a decreased oxygen consumption rate compared to that in wild-type cells. Basal, maximum, ATP-linked, and nonmitochondrial respiration rates were also reduced, and levels of ROS were increased.

In addition, in primary cultures of skeletal muscle, parkin KO cells were more sensitive (i.e., increased muscle cell death) than cells from wild-type animals to mitochondrial toxins including CCCP and rotenone, but no differences were observed in response to the more general toxins A23187 and H2O2 (Rosen et al., 2006). Parkin KO myofibers also had increased accumulation of oligomeric β-amyloid in cells exposed to Aβ42, reminiscent of pathology seen in patients with inclusion body myositis.

Finally, compared to wild-type mice, the sera of parkin KO mice (15 weeks of age) contained a greater amount of TOMM20, a marker of extracellular mitochondria (Choong et al., 2021).

Further reading on Mitochondrial Abnormalities
(full references listed at the end under Further Reading)

• Drew et al., 2014 (HSP72-parkin axis)
• Hoshino et al., 2013 (doxorubicin-induced mitochondrial abnormalities in heart tissue)
• Hoshino et al., 2014 (mitochondrial outcomes in pancreatic β-cells)
• Li et al., 2020 (mtDNA heteroplasmy)
• Li et al., 2022 (MFN2-mediated mitochondrial fusion and mitophagy)
• Lin et al., 2017 (mitochondrial docking via syntaphilin)
• Tostes et al., 2022 (NZB mtDNA accumulation in the liver)
• Trease et al., 2024 (double mutant cross with the Polg mitochondrial mutator line)
• Wang et al., 2011 (mitochondrial motility in cultured neurons)
• Williams et al., 2015 (mitochondrial function and mitophagy following alcohol-induced liver injury and steatosis)

Mitophagy

In primary cultured hippocampal axons, mitophagy initiation in response to antimycin A was absent in parkin KO cells based on the rate of mitochondrial colocalization with autophagosomes, while treatment of antimycin A induced mitophagy in wild-type axons (Ashrafi et al., 2014). Similarly, in proximal tubule cells of the kidney, rotenone-induced mitophagy was blocked in parkin KO mice but not in wild-type cells; however, basal mitophagy rates did not differ (Luciani et al., 2020). And when myocardial infarction was experimentally induced, parkin KO mice exhibited reduced mitophagy compared to wild-type controls, and the infarction also led to an accumulation of swollen, dysfunctional mitochondria in parkin KO but not wild-type mice (Kubli et al., 2013). In skeletal muscle, mitophagy was also abolished in response to acute endurance exercise in young parkin KO mice (Chen et al., 2018).

Further reading on Mitophagy Outcomes
(full references listed at the end under Further Reading)

• Alula et al., 2023 (Nix-mediated mitophagy and intestinal homeostasis)
• Araya et al., 2019 (mitophagy and cellular senescence outcomes in the lung following cigarette smoke exposure)
• Bhatia et al., 2019 (mitophagy in macrophages and kidney fibrosis)
• Chen et al., 2018 (exercise-induced mitophagy flux)
• Esteca et al., 2020 (impaired mitophagy during skeletal muscle regeneration)
• Kubli et al., 2015 (mitophagy pathways in cardiac myocytes)
• Lampert et al., 2019 (mitophagy in the differentiation of cardiac progenitor cells)
• Larson-Casey et al., 2016 (mitophagy and pulmonary fibrosis)
• Lee et al., 2011 (Bnip3-induced mitophagy in myocytes)
• Li et al. 2020 (mitophagy in renal fibrosis)
• Lin et al., 2019 (mitophagy and contrast-induced acute kidney injury)
• Wang et al., 2019 (acetaminophen-induced liver injury)
• Watzlawik  et al., 2021 (p-S65-Ub levels as mitophagy marker)
• Williams et al., 2015 (acetaminophen-induced liver injury)
• Yuan et al., 2017 (BNIP3L/NIX-mediated mitophagy and ischemic stroke)

Gastrointestinal Effects

At 12 to 14 weeks of age, parkin KO mice have impaired intestinal lipid absorption compared to wild-type control mice, based on the observations of increased fecal lipids and decreased plasma triglycerides after a short-term high-fat diet challenge (Costa et al., 2016). Nonetheless, small intestine morphology was normal in KO mice, and the expression of genes and proteins involved in intestinal lipid absorption also did not differ between wild-type and KO mice.

Metabolism

Changes in adipose tissue, a major metabolic organ, have been reported in parkin KO mice. For example, the transition from mitochondrial-rich, thermogenic beige adipocytes to mitochondrial-poor white adipocytes in response to the withdrawal of an external stimulus (β3-adrenergic receptor agonist CL316,243) was muted, with mutant mice retaining more beige adipocytes and showing reduced mitochondrial degradation compared to wild-type mice (Lu et al., 2018). 

In another study, thermogenic activity of brown adipose tissue was found to be overactivated in 3-month-old parkin KO mice based on the temperature of the interscapular region where brown adipose tissue is found (Cairó et al., 2019). Moreover, in response to cold exposure, parkin KO mice have over-induced expression of secretable thermogenesis-related factors. This same study also showed that when 3-month-old parkin KO mice were exposed to 8 weeks of a high-fat diet, they were resistant to weight gain and did not develop hyperinsulinemia.

Further reading on Metabolism:
(full references listed at the end under Further Reading)

• Kim et al., 2011 (metabolic outcomes following exposure to a high-fat and -cholesterol diet)
• Mooli et al., 2020 (mitochondrial biogenesis and the beige-to-white adipocyte transition following caloric restriction)
• Wang et al., 2021 (high fat diet–induced obesity)
• Wu et al., 2022 (high fat diet–induced metabolic, cardiac, and mitochondrial outcomes)
• Zhang et al., 2011 (glycolysis activation in mouse embryonic fibroblasts)

Protein Quality Control

Although parkin has E3 ubiquitin ligase activity, levels of the E3 ubiquitin ligase substrates CD-Crel-1, synphilin-1, and glycosylated α-synuclein were unchanged in parkin KO versus wild-type mice based on western blot analysis of brain tissue samples (Goldberg et al., 2003).

Levels of TDP-43, a protein involved in translation, were higher in the cortex of parkin KO mice compared to control mice (Wenqiang et al., 2014); this finding was also observed in 4- to 5-month-old parkin KO brain lysates compared to samples from controls (Hebron et al., 2014).

Protein levels of p62, which is a critical regulator of protein quality control, were selectively increased in the substantia nigra and striatum of 8-week-old parkin KO mice compared to wild-type mice; however, levels of p62 in other brain regions (hippocampus, frontal cortex, and cerebellum) were unaffected (Song et al., 2016).

Further reading on Protein Quality Control
(full references listed at the end under Further Reading)

• Cao et al., 2014 (endophilin)
• Dehvari et al., 2009 (phospholipase C-γ1)
• Ekholm-Reed et al., 2013 (Fbw7β)
• Ekholm-Reed et al., 2019 (Mcl-1)
• Gao et al., 2017 (CHIP–IRF1 axis)
• He et al., 2018 29987020 (PICK1)
• Kim et al., 2022 (saposin and cathepsin B)
• Lonskaya et al., 2013 (autophagic α-synuclein clearance)
• Lonskaya et al., 2014 (autophagic clearance of amyloid in lentiviral Aβ1–42-exposed mice)
• Matsuzawa-Ishimoto et al., 2017 (ATG16L1)
• Peng et al., 2020 (autophagy dysregulation following chronic alcohol exposure)
• Shires et al., 2020 (negative control for identifying ubiquitinated proteins in heart tissue)
• Sul et al., 2013 (Fas-associated factor 1)
• Watzlawik et al., 2024 (serine-65 phosphorylated ubiquitin)

Cellular Transport

Parkin KO mice exhibit abnormalities in microtubule stability in the dopaminergic terminals of the striatum, as measured in 2-, 7-, and 24-month-old mice, in comparison to wild-type mice (Cartelli et al., 2018). In addition, differences in mitochondrial clustering were observed in an age-dependent manner in parkin KO dopaminergic fibers: no differences in distribution were found between KO and wild-type at 2 months of age, but at 7 and 24 months, parkin KO mice had increased clustering, suggesting abnormalities in mitochondrial transport.

Levels of the retromer protein VPS35 did not differ between 3- to 4-month-old parkin KO and wild-type mice (Williams et al., 2018). However, levels of the WASH complex subunits WASH1 and FAM21 and the WASH-dependent retromer cargo ATG9A were decreased in the brains of parkin KO mice, suggesting perturbed endosomal sorting pathways.

Tumorigenesis, DNA Repair, and Cell Turnover

Mouse embryonic fibroblasts (MEFs) from parkin KO mice showed delays in key mitotic steps as well as increases in the protein expression of key mitotic regulators (Lee et al., 2015). In addition, more aneuploidy and polyploidy was observed in parkin KO cells, and they also escaped senescence.

In another study of MEFs from parkin KO mice, proliferation was accelerated, which corresponded to a greater the number of cells observed in S and M phases compared to cells from wild-type mice (Sarraf et al., 2019).

Protein expression of the tumor suppressor p53 is increased in fibroblasts derived from parkin KO mice, as is p53 activity, promoter transactivation, and mRNA levels (da Costa et al., 2009). Brain homogenates also show increased p53 protein and mRNA levels.

Parkin KO mice (18- to 20-week-old) had a lower incidence of lung tumor development in a urethane-induced carcinogenesis model compared to wild-type mice, as well as increased protein levels of the cell cycle arrester p21 (Park et al., 2019).

Further reading on Tumorigenesis, DNA Repair, and Cell Turnover
(full references listed at the end under Further Reading)

• Aksoy Yasar et al., 2022 (tumorigenicity in a double mutant cross with conditional Trp53 and Pten deletions)
• Kao 2009 (DNA repair in embryonic stem cells)
• Li et al., 2018 (pancreatic tumorigenesis)
• Zhu et al., 2017 (susceptibility to ultraviolet radiation)

Immune Function

Signs of inflammation and hyperplasia were observed in 10-month-old parkin KO mice, including inflammatory infiltration in the small intestine, increased frequency of rectal prolapse, splenomegaly, and increased stool softness and blood in the stool (Lee et al., 2019). In addition, levels of inflammatory markers (TNF-α, IL-1β, and IL-6) were elevated in the small intestine and in MEFs compared to wild-type mice. At 12 months of age, parkin KO mice also had an increased number of polyps in the small intestine.

Parkin KO mice exposed to Pseudomonas aeruginosa to model pneumonia had a reduced capacity to kill the bacteria (Bone et al., 2021). Similarly, parkin KO mice were also highly susceptible to infection by Listeria monocytogenes or Mycobacterium tuberculosis, and had much more difficulty than wild-type mice in controlling these infections (Manzanillo et al., 2013). In a model of prion disease, intracranial RML prion infection resulted in worse survival in parkin KO versus wild-type mice (Ward et al., 2024).

Following lipopolysaccharide treatment, peritoneal macrophages from parkin KO mice have an exacerbated increase in the mRNA expression of some inflammatory genes (TNF, IL-1b, and iNOS), but not others (Nrf2, HO-1, and NQO1) compared to cells from wild-type mice (Tran et al., 2011).

Innate antiviral immunity is enhanced in parkin KO MEFs compared to wild-type cells in response to two RNA viruses (Sendai virus and vesicular stomatitis mutant virus) and two DNA virus triggers (interferon stimulatory DNA and herpes simplex virus 1; Xin et al., 2018).

Further reading on Immune Function
(full references listed at the end under Further Reading)

• Abuaita et al., 2018 (mitochondria-derived vesicles in response to methicillin-resistant Staphylococcus aureus)
• de Carvalho et al., 2019 (Leishmania RNA virus)
• Kinsella et al., 2023 (Mycobacterium tuberculosis)
• Lee et al., 2016 (inflammation in the lung and genomic instability)
• Matheoud et al., 2016 (mitochondrial antigen presentation)
• Matheoud et al., 2019 (mitochondrial antigen presentation)
• Sliter et al., 2018 (exhaustive exercise–induced inflammation)
• Smith et al., 2019 (influenza A virus and macrophage migration inhibitory factor)

Cardiac Function

The heart to body weight ratio of 8- to 10-week-old parkin KO mice did not differ from that of wild-type control mice (Kubli et al., 2013; also see Woodall et al., 2019). In addition, echocardiography findings (fractional shortenings, ejection fractions, left ventricular internal end diastolic and systolic dimensions) did not differ between KO and wild-type mice at 3, 6, or 12 months of age. However, experimentally induced myocardial infarction led to greater mortality in parkin KO versus wild-type mice, which was accompanied by more severe histologic and echocardiographic outcomes.

Further reading on Cardiac Function
(full references listed at the end under Further Reading)

• Han et al., 2017 (outcomes in response to cardiac pressure-overload or tunicamycin)
• Leach et al., 2017 (outcomes following myocardial infarction in a double mutant cross with a conditional knockout of Salv mouse)
• Tong et al., 2019 (cardiac dysfunction in response to a high-fat diet)
• Tyrrell et al., 2020 (aortic mitochondrial function following IL-6)
• Ma et al., 2021 (TRAF2 ablation and cardiomyopathy)
• Wang et al., 2018 (melatonin effects on cardiac remodelling in diabetic cardiomyopathy)
• Wang et al., 2019 (diabetic cardiomyopathy)

Other
(full references listed at the end under Further Reading)

Further reading (Retinal Outcomes):
• Chen et al., 2013 (light-induced retinal degeneration)
• Chen et al., 2019 (retinal injury outcomes in triple knockout SOD1/DJ-1/Parkin mice)
• Chen et al. 2020  (retinal degeneration in triple knockout SOD1/DJ-1/Parkin mice)
• Ding et al., 2017 (light-induced retinal degeneration in triple knockout SOD1/DJ-1/Parkin mice)
• Foster et al., 2018 (manganese exposure on various physiological outcomes)
• Kim et al., 2023 (osteoclast activity and bone mass)
• Zhu et al., 2019 (spontaneous retinal degeneration with aging in triple knockout SOD1/DJ-1/Parkin mice)

Further reading (Miscellaneous):
• Li et al., 2022 (aminoglycoside-induced hearing loss)
• Lin et al., 2023 (cisplatin-induced acute kidney injury)
• Liu et al., 2017 (mitochondrial function in the ovary)
• Mizumura et al., 2014 (airspace enlargement following chronic cigarette smoke exposure)
• Wu et al., 2023 (aggravated CCl4-induced liver fibrosis)
• Zhang et al., 2021 (hyperoxia-induced lung injury)

Modification Details

This KO mouse model was generated by homologous recombination in J1 (129/Sv) embryonic stem cells, which were injected into C57BL/6 and Balb/c blastocysts (Goldberg et al., 2003). Chimeric mice were further crossed to C57BL/6 mice to establish germline transmission, and backcrossed to C57BL/6 for more than 20 generations before becoming commercially available (The Jackson Laboratory). The targeted exon 3 was replaced with part of exon 3 fused in-frame to EGFP followed by a stop codon. The resulting transcript codes for the first 95 amino acids of mouse Parkin, followed by the full EGFP amino acid sequence. Northern analysis also revealed a transcript that skips the mutant exon 3, leading to a frameshift mutation after the first 57 amino acids of mouse Parkin and premature termination after out-of-frame translation of amino acids 58-105.

Phenotype Characterization

Q&A with Model Creator

Q&A with Matthew Goldberg

What would you say are the unique advantages of this model?
The in-frame EGFP can be detected by western analysis and immunohistochemistry. Thus, EGFP can be used as a reporter for parkin expression both spatially and in experiments evaluating treatments that alter parkin expression.

What do you think this model is best used for?
Biochemical studies of the effects of inactivation of parkin in vivo.

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References

Paper Citations

1. Goldberg MS, Fleming SM, Palacino JJ, Cepeda C, Lam HA, Bhatnagar A, Meloni EG, Wu N, Ackerson LC, Klapstein GJ, Gajendiran M, Roth BL, Chesselet MF, Maidment NT, Levine MS, Shen J. Parkin-deficient mice exhibit nigrostriatal deficits but not loss of dopaminergic neurons. J Biol Chem. 2003 Oct 31;278(44):43628-35. Epub 2003 Aug 20 PubMed.
2. Kitada T, Asakawa S, Hattori N, Matsumine H, Yamamura Y, Minoshima S, Yokochi M, Mizuno Y, Shimizu N. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature. 1998 Apr 9;392(6676):605-8. PubMed.
3. Hollville E, Joers V, Nakamura A, Swahari V, Tansey MG, Moy SS, Deshmukh M. Characterization of a Cul9-Parkin double knockout mouse model for Parkinson's disease. Sci Rep. 2020 Oct 9;10(1):16886. PubMed.
4. Palacino JJ, Sagi D, Goldberg MS, Krauss S, Motz C, Wacker M, Klose J, Shen J. Mitochondrial dysfunction and oxidative damage in parkin-deficient mice. J Biol Chem. 2004 Apr 30;279(18):18614-22. Epub 2004 Feb 24 PubMed.
5. Moore TM, Cheng L, Wolf DM, Ngo J, Segawa M, Zhu X, Strumwasser AR, Cao Y, Clifford BL, Ma A, Scumpia P, Shirihai OS, Vallim TQ, Laakso M, Lusis AJ, Hevener AL, Zhou Z. Parkin regulates adiposity by coordinating mitophagy with mitochondrial biogenesis in white adipocytes. Nat Commun. 2022 Nov 4;13(1):6661. PubMed.
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External Citations

1. The Jackson Laboratory
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