5 questions

Brain stuff

eEF1BSL = research interest

—> one gene two proteins Alternative: already know about Cassette exon: include or exclude the middle exon; two types of proteins are still produced Last one: the exon is retained or spread??

Second one = cassette exon splicing - include or exclude type Leads to long form (left) or short form (right)

EIFs = eukaryotic [] factor El

EEf1A = rich in cells??? Today’s = the third one (delta one) The last one is just the structural protein of the above products; supports protein translation

Shows how the protein was produced through the elongation step; basically just address that he was trying to say that it supports protein translation

Western blot showed that the long form is only present in brain and testes

Left one (delta 1) : Gfp localized to cytoplasm Right one (delta L): gfp localized to cytoplasm and the nucleus

Left side is original, as shown, localized to cytoplasm and nucleus; mutant on right is only localized to the (nucleus???) by its NLS region.

Role in transcription HSE increase shown in dL showed that heat shock showed up ______. 14:17 on rec.

Here shows heat shock binds to dL protein in ___???

Basically all we can say from that is that DL protein has a role in heat shock by method of alternative splicing

Physiological Applications

Incorporated biochemical analysis with behavioral analysis to accurately depict the neurological phenotype

Shows where expression exists; showed that they succeeded in __

Used mazes to test anxiety and responses. The mice tended to stay in the closed arm section of the elevated plus maze. Compared to the wild type, there was no significant difference in anxiety.

Contextual task was Cued task: only tone was applied applied at 2, 4, and 6 minutes; was found that increased freezing time as a result of this. After 3 minutes, freezing increased (at 4 minute time increment) [video shown 1: where mouse was normally moving in circles around field until the shock was applied when the mouse jumped and accelerated still running around in same pattern 2: all of the above but collapsed and had a seizure afterwards]

Shows # that had a seizure —few.

To keep variables constant, mice were maintained under same conditions; variables were monitored at the same time each day and the mice lived under the same light/ dark cycles. Right graph shows that the wild type mouse was more active overall; much more in dark time and a little bit more during light time. Bottom left shows how the KO mouse had suppressed locomotive activity throughout.

g dosed

Papers that he found on pubmed show answers to main question posed by this study. Left one showed that aversive stimuli could be used to reduce ptsd in mice. Right one showed same thing??? Basically just gave him better insight to seeing why his mice were showing suppressed locomotive activity.

Imaging from dissected brains from his mice. What he found: Unchanged body mass but brain weight was significantly reduced in the KO??? mouse ^he said the wt mouse but graph show knockout

Neuroprotective gene: ie heat shock

Gene type was unchanged between the viral type ad th KO type So he thinks it was the result of another biochemical phenotype (shown: other phenotypes tested) Balance was changing....

How did he really test this change in “balance” Found: basal protein synthesis was increased in S labeling and puromycin labeling Basically it increased.

So...this is how it resulted in brain atrophy. ...but why seizures? (Next slide he just showed tha the looked it up)

Now we think?? 41:02

Found some research for implications on psychiatric disorders Ex of the disorders studied: autism, etc. In this study he looked at, it showed that there was a 69 deletion; basically this mutation made the patient not have the dL protein!!!! Much cool. So wow.

Shows that this this has real human implications, illustrates the importance of everything ^^.

Protein Transduction

The basics of protein transduction. HIs PI discovered? protein transduction —> if this is true that’s cool. <Much cool. So wow.>

Arginine can enable __11R Viral type not found w/ EGFP.

The how: methods for this research of his and what they mean. Shows that there was a “robust” florescence in the cell with the 11R but not the EGFP.

Commercially produced staining on E. Coil. Max effect shown on cell internalization. Using something that sounded like Alexa....49:37

[missed a slide that he glazed over further describing ^]

Showed that many insulin producing cells were differentiated with

Just in case you’re tryna cite people in your paper.

**51:00 onwards is Q&A. Important questions were asked that I didn’t take notes on.