Mouse anti-Human Tom20 monoclonal antibody, clone 30/Upn30

Contextual fear conditioning (CFC) is a quick cognitive test based on the association context-aversive stimulus in which a single training leads to a long-term memory. Previously, we analyzed the gene expression in CFC rats, naive rats, explor rats which had freely explored the experimental apparatus and shock only (SO) rats to which the same number of aversive shocks used in the CFC paradigm had been administered in the same CFC apparatus in less time to prevent the association between painful stimuli and apparatus. The analysis showed that 2 days after conditioning some genes (Napa, Pnf2, Casp3, Pdrg1, Ywhaz, Stmn1, Bpgm) were more expressed in CFC rats with respect to naive, explor and SO rats. Other genes (Actr3, Pea15,Tiprl) were more expressed in SO rats or in explor rats (Cplx1, Trim32, Ran), and the expression of the Tomm20 gene was greater in both CFC and explor rats in comparison with SO and naive rats. Herein, we have tested the expression of these genes for longer periods, at 3, 10 and 20 days after conditioning. The expression of the transcripts was assessed by qRT-PCR. Interestingly, in CFC rats, at 3 days, the genes Tiprl and Trim32, whose expression had not been modulated at 2 days resulted more expressed with respect to naive, explor and SO rats, whereas the gene Tomm20 was less expressed as in the SO and explor rats with respect to naive rats. At 10 days, Trim32 gene was still more expressed in CFC rats whereas the genes Tiprl and Tomm20 returned to the constitutive level, and the gene Ran was significantly more expressed in CFC rats than in naive, SO and explor rats. Surprisingly at 20 days, the genes Sunni and Tiprl again resulted significantly more expressed in CFC rats compared with naive, SO and explor rats. Our results outline a complex modulation of gene expression in a prolonged time point of CFC post-acquisition consolidation period.

This study aimed to determine if there is an association between mitophagy and refractory temporal lobe epilepsy (rTLE) with hippocampal sclerosis. During epilepsy surgery, we collected tissue samples from the hippocampi and temporal lobe cortexes of rTLE patients with hippocampal sclerosis (as diagnosed by a pathologist). Transmission electron microscopy (TEM) was used to study the ultrastructural features of the tissue. To probe for mitophagy, we used fluorescent immunolabeling to determine if mitochondrial and autophagosomal markers colocalized. Fourteen samples were examined. TEM results showed that early autophagosomes were present and mitochondria were impaired to different degrees in hippocampi. Immunofluorescent labeling showed colocalization of the autophagosome marker LC3B with the mitochondrial marker TOMM20 in hippocampi and temporal lobe cortexes, indicating the presence of mitophagy. Mitochondrial and autophagosomal marker colocalization was lower in hippocampus than in temporal lobe cortex (P < 0.001). Accumulation of autophagosomes and mitophagy activation are implicated in rTLE with hippocampal sclerosis. Aberrant accumulation of damaged mitochondria, especially in the hippocampus, can be attributed to defects in mitophagy, which may participate in epileptogenesis.