Postembryonic development of serotonergic neurons in the abdominal nervous system of the mealworm Zophobas morio (Coleoptera: Tenebrionidae)

2011 Research primer 2

The wall of the hindgut of the larval mealworm, Zophobas morio. A dense meshwork of the branches of axons containing 5HT-like immunoreactivity covers the smooth muscle
Muscle fibers

hindgut

From midgut

To rectum & anus

axons in proctodeal nerves

Method: Immunocytochemistry
Use anti-serotonin antibodies to bind to serotonin To this, bind a secondary antibody that as a fluorescent tag Visualize using a fluorescent microscope
Serotonin

Rabbit Anti-serotonin

Goat antirabbit with CY3 fluorophore

The serotonergic axons come from neurons in the terminal abdominal ganglion
hindgut

proctodeal nerves

Terminal abdominal ganglion

200 m

Expression of serotonin-like immunoreactivity Zophobas. Terminal abdominal ganglion. Larva

axons of the interneurons, ascending to other ganglia (3/side) cell bodies of interneurons (3/side)
cell bodies of motor neurons (2/side) axons of the motor neurons, going to the hindgut (2/side)

100 m

Expression of serotonin-like immunoreactivity Zophobas. Terminal abdominal ganglion. Pupa, 1d

Terminal abdominal ganglion

Abdominal ganglion VIII

Abdominal neuromeres:

IX X

XI

Expression of serotonin-like immunoreactivity Zophobas. Terminal abdominal ganglion. Pupa, 10d

Terminal abdominal ganglion

Abdominal neuromeres:

VII VIII IX X

XI

Questions arising
If this developmental description holds up: Why does the pattern of serotonergic neurons change during metamorphosis? Ideas: 1. Some new neurons are born; some old ones die. 2. Some existing neurons change their ability to synthesize serotonin. 3. These changes are controlled by developmental hormones.

Biosynthetic pathways of serotonin and dopamine

TPH

DDC

TH

a.k.a Dopa decarboxylase, DDC

Expression of TPH-like immunoreactivity

Zophobas, Terminal abdominal ganglion. Larva
lateral anterior
extra neurons (5/side)

100 m

Comparison of serotonin- and TPH-like immunoreactivity: larva-to-larva

TPH, larva 5HT, larva

IX X XI X XI

IX

Comparison of serotonin- and TPH-like immunoreactivity: larva-to-pupa

TPH, larva
VIII TPH, larva

5HT, pupa

VII

5HT, larva
VIII

IX X XI XI

IX
X

Tentative hypotheses, further questions

Maybe the new 5HT-neurons of the late pupa are already present in the larva and express TPH, but not DDC. In other words, the serotonergic capability of neurons is acquired in developmental stages. But are there neurons that express TPH-like but never 5HT-like IR?

This summer: confirming the developmental story, testing hypotheses

1. Establish developmental timecourse of 5HTlike IR in the terminal abdominal ganglion. 2. Follow TPH-like IR and DDC-like IR thru the same sequence. 3. Double labeling? 4. Controls

General pattern of serotonergic neurons in a free abdominal ganglion. Zophobas, larva

lateral anterior

2 interneurons per side