Effect of visual experience on

ventral-preferring DSGC patterning

in the mouse retina

Kayla Maanum
Professional Audience Piece
May 10, 2019
The retina is a highly organized piece of neural
tissue located in the back of the eye.
Mouse Eye

Retinal ganglion cells (RGCs) are

neurons whose axons comprise
the optic nerve.

optic nerve

There are subpopulations of

RGCs that preferentially fire
action potentials in response to
certain features of the visual
scene.

(Beier et al., 2013)

 Dendrites of cells within a subtype repel each
other in development, forming soma mosaics.

dendritic repulsion
allows equal
spacing between
cells of the same
type (Rivlin-Etzion et al., 2011)

We are interested in a subtype of RGCs that encodes directional motion. These cells are
known as ON-OFF direction-selective ganglion cells (ON-OFF DSGCs).
 Four subtypes of ON-OFF DSGCs that each
prefer one of four cardinal axes of visual motion

four cardinal axes:

Left: Somata of DSGCs are encircled. Right: Tuning curves

for cells of four subpopulations.

(Morrie and Feller 2016)

Ventrally-tuned ON-OFF DSGC (Hb9-DSGC)
dendrites are morphologically asymmetric.
 When deprived of visual experience, adult Hb9-
DSGCs exhibit more symmetric dendritic
structure. Control Dark Reared
12 hr light/dark cycle 24 hour dark cycle

Eye Opening
P13-14

Adult
P>30
Morphological change in dark-reared Hb9-DSGC
dendrites aligns with ontogenetic timeline.

Dark-reared animals are

housed in 24-hr dark cycle
from birth.
Before eye opening, the retinal
circuit is immature and not fully
connected.
 Morphological change in dark-reared Hb9-DSGC
dendrites aligns with ontogenetic timeline.

Dark
Reared
 Visual deprivation prevents DSGC tuning
curves from clustering at four cardinal axes.

Eye Opening

(Bos et al. 2015)

Research Questions:

1. How does visual experience affect the area of Hb9-DSGC

dendritic arbor?

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes?

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in normally-developed adults?
Research Questions:

1. How does visual experience affect the area of Hb9-DSGC

dendritic arbor?

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes?

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in adulthood?
Maximum intensity z-stack projections of dye-filled
Hb9-DSGCs analyzed on ImageJ.

In ImageJ: draw convex ROI around farthest extent of

dendrites.
 Visual deprivation increases dendritic field area
by adulthood.

In ImageJ: draw convex ROI around farthest extent of Unpaired t-test Adults: p=0.1745; Eye opening:
dendrites. Calculate area. p=0.0069**
Questions:

1. How does visual deprivation affect the area Hb9-DSGC

dendritic arbor? → Increases dendritic field size by adulthood.

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes?

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in adulthood?
Research Questions:

1. How does visual deprivation affect the area Hb9-DSGC

dendritic arbor? → Increases dendritic field size by adulthood.

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes?

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in adulthood?
Immunohistochemical staining can be used to
label Hb9-DSGC somata on a whole retina.
Immunohistochemical staining can be used to
label Hb9-DSGC somata on a whole retina.

Each dot is the soma of a

single Hb9-DSGC.
Custom MATLAB script determines nearest
neighbor distance between each pair of Hb9-DSGC
somata.
Custom MATLAB script determines nearest
neighbor distance between each pair of Hb9-DSGC
somata.

Masks of somata created for each stained Smallest pairwise distance (red) calculated for
retina. each cell on retina.
Nearest neighbor distance between Hb9-DSGC
somata retained in absence of visual experience.

Eye Opening Adult

Research Questions:

1. How does visual deprivation affect the area Hb9-DSGC

dendritic arbor? → Increases dendritic field size by adulthood.

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes? → No; somata patterning is independent of dendritic development.

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in adulthood?
Research Questions:

1. How does visual deprivation affect the area Hb9-DSGC

dendritic arbor? → Increases dendritic field size by adulthood.

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes? → No; somata patterning is independent of dendritic development.

1. Is gap junction coupling a possible mechanism for refined

DSGC tuning in adulthood?
Neurobiotin can pass through gap junctions into
somata of coupled cells.

Patch clamp technique used to perfuse

Neurobiotin+ AlexaFluor594 dye into Hb9-
DSGC soma, which then perfuses into
somata of coupled cells.

somata of coupled cells

Colocalization imaging on confocal reveals
homologous vs. heterologous coupling.

Hb9-DSGCs Filled and coupled Arrowhead (yellow): homologous

cells coupling between cells of the same
type. Asterik (red): heterologous
coupling between cells different type.
 Preliminary data suggest no differences in
identities of coupled cells between control and
dark-reared adults.

Contro
l
P13-14

Contro
l
Adult

Dark
Adult
Research Questions:

1. How does visual deprivation affect the area Hb9-DSGC

dendritic arbor? → Increases dendritic field size by adulthood.

1. Does visual experience affect mosaic patterning of Hb9-

DSGC somata on the retina as a result of dendritic area
changes? → No; somata patterning is independent of dendritic development.

1. Is gap junction coupling a mechanism for refined DSGC

tuning in adulthood? → To be determined.
 Future Research

• Are morphological changes to dendrites a

secondary result to other deprivation-
mediated changes in the IPL?
• Are dendritic field areas of On and Off
dendrite layers separately affected?
• How do gap junction coupling identities
change during development?
 References

Beier, K., Borghuis, B., El-danaf, R.,Huberman, A., Demb, J & Cepko, CL. (2013). Transsynaptic Tracing with
Vesicular Stomatitis Virus Reveals Novel Retinal Circuitry, The Journal of Neuroscience, 33: 35-51.
Morrie, R.D. and Feller, M.B. (2018). A Dense Starburst Plexus Is Critical for Generating Direction Selectivity,
Current Biology, 28(8):1204-1212.
Rivlin-Etzion, M, Zhou, K, Wei, W, Elstrott, J, Nguyen, P, Barres, B, Huberman, A, Feller, M. (2011).Transgenic
mice reveal unexpected diversity of On-Off direction selective retinal ganglion cell subtypes and brain
structures involved in motion processing. Journal of Neuroscience 2011, 31(24):8760-9.
 Acknowledgements

Thank you!
Malak El-Quessny Daniela
Kaufer
Marla Feller Caroline
Williams

Feller Lab including:

Alex Tiriac
Christiane Vuofo
Mathew Summers Kimberly
Cachero
Franklin Caval-Holme Joshua Tworig
 Acknowledgements (continued)

Kim Freeman
College Writing 161 class

Departments of Integrative Biology and MCB