An Alternatively Spliced TREM2 Isoform Lacking the Ligand Binding Domain is Expressed in Human Brain.

BACKGROUND: Genetic variants in TREM2 are strongly associated with Alzheimer's disease (AD) risk but alternative splicing in TREM2 transcripts has not been comprehensively described. OBJECTIVE: Recognizing that alternative splice variants can result in reduced gene expression and/or altered function, we sought to fully characterize splice variation in TREM2. METHODS: Human anterior cingulate autopsy tissue from 61 donors was used for end-point and quantitative PCR and western blotting to identify and quantify novel TREM2 isoforms. RESULTS: In addition to previously described transcripts lacking exon 3 or exon 4, or retaining part of intron 3, we identified novel isoforms lacking exon 2, along with isoforms lacking multiple exons. Isoforms lacking exon 2 were predominant at approximately 10% of TREM2 mRNA in the brain. Expression of TREM2 and frequency of exon 2 skipping did not differ between AD samples and non-AD controls (p = 0.1268 and p = 0.4909, respectively). Further, these novel splice isoforms were also observed across multiple tissues with similar frequency (range 5.3 -13.0%). We found that the exon 2 skipped isoform D2-TREM2 is translated to protein and localizes similarly to full-length TREM2 protein, that both proteins are primarily retained in the Golgi complex, and that D2-TREM2 is expressed in AD and non-AD brain. CONCLUSION: Since the TREM2 ligand binding domain is encoded by exon 2, and skipping this exon retains reading frame while conserving localization, we hypothesize that D2-TREM2 acts as an inhibitor of TREM2 and targeting TREM2 splicing may be a novel therapeutic pathway for AD.

An active avoidance behavioral paradigm for use in a mild closed head model of traumatic brain injury in mice.

BACKGROUND: A mild traumatic brain injury (TBI) occurs to millions of people each year. Translational approaches to understanding the pathogenesis of neurological diseases and the testing of the effectiveness of interventions typically require cognitive function assays in rodents. NEW METHODS: Our goal was to validate the active avoidance task using the GEMINI avoidance system in a mouse model of mild closed head injury (CHI). RESULTS: We found that shock intensity had only a marginal effect on the test. We found that sex was an important biological variable, as female mice learned the task better than male mice. We demonstrate that a single mild CHI in mice caused deficits in the task at four weeks post-injury. COMPARISON WITH EXISTING METHODS: Active avoidance is a classical conditioning test in which mice must pair the presence of a conditioned stimulus with moving between two chambers to avoid an electric shock. External conditions (i.e., apparatus), as well as inherent differences in the mice, which may not be directly linked to the model of the disease (i.e., sensory differences), can affect the reproducibility of a behavioral assay. Before our study, there was a lack of standard operating procedures and validated methods for the active avoidance behavior for phenotyping mouse models of injury and disease. CONCLUSION: We offer a method for validating the active avoidance test, and a standard operating procedure, which will be useful in other models of neurological injury and disease.