Class Project: Postsynaptic Resources¶

NEU466G, Dr. Harris · Class documents

Overview¶

Weeks 8 and 9 of the class research project. In every spine of your group's dendrite, look for four kinds of postsynaptic resources and place a stamp on each one you find:

smooth endoplasmic reticulum (SER)
spine apparatus (SA)
polyribosomes (PR)
endosomes

You already know how: this is the same work as Lab 3.1 and Lab 3.2, on more spines and on a new dendrite.

Where to look: the head and the neck of every spine on your group's dendrite.
What you make: one stamp for each SER, SA, polyribosome cluster or endosome you find, named after its spine.
What you hand in: your saved series, uploaded to Canvas at the end of each class.

What each resource looks like¶

Go back to the labs for the descriptions, the example images and the papers:

SER: Lab 3.1, Smooth endoplasmic reticulum.
SA: Lab 3.1, Spine apparatus.
Endosomes: Lab 3.1, Endosomes, with the criteria for each of the four types.
Polyribosomes: Lab 3.2, Identifying polyribosomes.

The example images in the labs come from the starter series (ZGBJY). The tissue in your series looks much the same.

Names for your stamps¶

In the labs, each name was the dendrite, then the resource, then the spine number: d03ser12 is SER in spine 12 of dendrite d03. The project uses a naming scheme of the same kind, so the class data can be put together.

Resource	Name in the labs	Name in the project
SER	`d03ser12`	To be decided
SA	`d03sa12`	To be decided
Polyribosomes	`d03r12`	To be decided
Endosome (coated pit)	`d03endo12_cp`	To be decided
Endosome (other types)	none	To be decided

To be decided

The naming scheme for the project: the dendrite part, whether a location letter is added (h for head, n for neck, b for base, s for shaft, as in the example stamps of the starter series), and how to name the two branches of a branched spine.
Names for endosomes other than coated pits (multivesicular bodies, tubules, vesicles), or whether to stamp coated pits only.
The colors and the palette squares, if they are set up in the series for you.

Stamp the resources¶

Use the same steps as in the labs. For each name:

In the Tools window, select the Stamp tool.
Right-click an unused palette square. The Set Attributes dialog opens.
In Name, type the name for what you found.
In Stamp radius (microns), type 0.05.
Click OK.
Click on the SER, SA, endosome or polyribosome cluster to place the stamp.

Use a separate palette square for each name. Two rules from the labs still apply:

SER and SA: stamp any SER and SA you see in the spine head and the spine neck (Lab 3.1, Stamp spine SER and SA).
Polyribosomes: stamp each cluster on just one section where it is present. If a spine has more than one cluster, stamp each cluster once, all with the same name (Lab 3.2, Stamp polyribosomes).

Check your stamps¶

In the Object List, choose Filter > Regex filter... and type a filter that lists your stamp names, as in Lab 3.1, Check your stamps.
Check that every stamp has the right name for its spine.
To see the polyribosomes as spheres in 3D, follow Lab 3.2, Adding polyribosomes to the 3D scene.

To be decided

How you record the resources for the class data in Week 10 (for example, counts per spine in a shared spreadsheet). See Class data.

Saving¶

At the end of each class, save with Ctrl+S (Mac: Cmd+S), back up your .jser file, and upload it to Canvas, as in Lab 1.1.