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. 2020 Jul 31;13:144. doi: 10.3389/fnmol.2020.00144

TABLE 1.

Summary of apoER2 alternatively spliced exons, including their associated in vitro and in vivo phenotypes, spatiotemporal distribution and known splicing regulators.

Alternative Exon Functional domain affected In vitro phenotype In vivo phenotype Spatiotemporal distribution Splicing regulator Associated publications
Murine and Human exon 5 LDLa ligand binding repeats 4–6 Required for α2-macroglobulin binding (HEK-293 cells) Unknown Murine:
– solely Δex5 detected (total brain RNA)
Human:
– +/Δex5 detected (total brain RNA)
– solely Δex5 detected in adult frontal cortex, hippocampus and cerebellum and fetal brain (RNA)
Unknown Kim et al., 1997; Clatworthy et al., 1999; Brandes et al., 2001
Murine exon 7 LDLa ligand binding repeat 8 (lacking in humans) – Exclusion leads to lower affinity for β-VLDL particles (HEK-293 cells)
– Exclusion confers altered affinity for reelin cleavage products (COS7 cells)
Unknown – +/Δex7 both detected (total brain RNA)
– Δex7 only at E10 (total brain RNA)
– +/Δex7 at E12 and E15/16 (total brain RNA)
– E15/16: cerebrum contains +/Δex7 while cerebellum and olfactory bulbs soley express +ex7 (RNA)
– Ex7 cerebrum expression higher at E15-17 than P10 (protein)
– Ex7 cerebellar expression increases from E17 to P10 (protein)
– Cerebellum contains higher ex7 levels than cerebrum at P10 (protein)
– P10 cerebellum: ex7 expression in internal granule cell layer but not external granule layer or Purkinje cells (protein)
– E17 ex7 expression in cortical plate, but scarce in Marginal, Intermediate and Subventricular zones (protein)
Unknown Brandes et al., 1997, 2001; Kim et al., 1998; Koch et al., 2002; Hibi et al., 2009
Murine exon 7B
(Human exon 6B)
Furin cleavage site – Cleaved in furin dependent manner (HEK-293 cells)
– Extracellular cleavage product binds reelin
– Cleavage product suppresses Dab1 phosphorylation (primary neurons IP)
Cleavage product remains undetected in brain Murine:
– low level inclusion of ex7B (total adult and embryonic brain RNA)
– mutually exclusive with ex7 (total brain RNA)
– low level inclusion of ex7B in E15-16 cerebrum but none in E15-16 cerebellum or olfactory bulb (RNA)
Human:
– low level inclusion of ex6B (total brain RNA)
Preliminary evidence for Rbfox2 Brandes et al., 1997, 2001; Koch et al., 2002; Gehman et al., 2012
Murine exon 16
(Human exon 15)
O-linked glycosylation site; extracellular cleavage site – Required for extracellular cleavage by metalloproteases (HEK-293 cells and primary neurons)
– Required for receptor sugar glycosylation (CHO cells and brain lysate)
– Exclusion alters hippocampal LTP and CA1 synaptic strength in exon 19 dependent manner (hippocampal slice electrophysiology)
Murine:
Constitutive exclusion leads to:
– increased brain apoER2 mRNA and protein levels
– increase in hippocampal dendritic spines
– impaired fear acquisition (context and cued fear conditioning task)
– augmented glutamate receptor expression
Human:
– +/Δex15 both detected in adult frontal cortex, hippocampus and cerebellum and fetal brain (RNA)
Unknown Clatworthy et al., 1999; Wasser et al., 2014
Murine exon 19
(Human exon 18)
Cytoplasmic insert – Binds adaptor proteins JIP1, JIP2, PSD-95, APBA1, APBA2 (Y2H screen, IP)
– Required for reelin-induced LTP in hippocampus (hippocampal slice electrophysiology)
Murine:
Ex19 inclusion lower in aging and amyloid mouse model hippocampus (semi-qRT-PCR)
Constitutive exclusion confers:
– deficits in hippocampal dependent learning (fear conditioning/associative learning)
– decreased number of adult corticospinal neurons
– decreased thickness of adult primary motor cortex
– protection from corticospinal neuronal death following brain injury
Human:
– Ex18 inclusion is lower in middle temporal region of the AD brain (semi-qRT-PCR)
– Ex18 inclusion associated with global cognition (linear regression models)
Murine:
– brain stem cells solely express Δex19
– approximately equal +/Δex19 levels found in primary neurons, total brain RNA and hippocampal RNA
– ex19 confirmed at protein level (total brain lysate)
Human:
– approximately equal +/Δex18 levels in adult middle temporal region, frontal cortex, hippocampus and cerebellum and whole fetal and adult brains (RNA)
– Moderate evidence for SRSF1
– Activity regulated
Kim et al., 1998; Clatworthy et al., 1999; Stockinger et al., 2000; Gotthardt et al., 2000; Beffert et al., 2005; Beffert et al., 2006; He et al., 2007; Hinrich et al., 2016

LDLa, Low density lipoprotein receptor class A; HEK-293, Human Embryonic Kidney-293; VLDL, Very low density lipoprotein; IP, immunoprecipitation; LTP, Long term potentiation; Y2H, Yeast-2-Hybrid; qRT-PCR, quantitative real time-polymerase chain reaction.