# Differentiation timing-dependent axon targeting and subtype specification in retinal ganglion cells

**Authors:** Lena Iwai, Tatsumi Hirata

PMC · DOI: 10.3389/fnins.2026.1733811 · Frontiers in Neuroscience · 2026-02-10

## TL;DR

This study shows that the timing of retinal ganglion cell development influences their specialization and axon targeting in the mouse retina.

## Contribution

The study demonstrates a novel method to tag and analyze RGC subtypes based on their differentiation timing.

## Key findings

- RGC axon projections to specific brain regions depend on differentiation timing.
- Neurogenically tagged RGCs show distinct molecular and morphological features.
- Differentiation timing is linked to RGC subtype specification.

## Abstract

Subtypes of retinal ganglion cells (RGCs) in the mouse retina are each tuned to particular visual features and contribute to parallel visual processing in the brain. We addressed how RGCs are specified into distinct and diverse subtypes based on their differentiation timing. We used a neurogenic tagging mouse line, Neurod1CreER (D1B), in which tamoxifen-inducible CreER was driven by a putative Neurod1 enhancer. Timed tamoxifen injection in this mouse line induced CreER-loxP recombination in neurons that shared the same differentiation timing. This analysis revealed that RGC axon projections to the lateral geniculate nucleus and medial terminal nucleus were segregated depending on the stage of tamoxifen injection. We further characterized the properties of these neurogenically tagged RGCs based on their molecular markers and morphological features. Our study extends the concept that differentiation timing is linked to the specification of RGC subtypes.

## Linked entities

- **Genes:** NEUROD1 (neuronal differentiation 1) [NCBI Gene 4760]
- **Chemicals:** tamoxifen (PubChem CID 2733526)
- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** Atoh7 (atonal bHLH transcription factor 7) [NCBI Gene 53404] {aka Math5, bHLHa13}, Thy1 (thymus cell antigen 1, theta) [NCBI Gene 21838] {aka CD90, T25, Thy-1, Thy-1.2, Thy1.1, Thy1.2}, Opn4 (opsin 4 (melanopsin)) [NCBI Gene 30044] {aka 1110007J02Rik, Gm533}, MTFP1 (mitochondrial fission process 1) [NCBI Gene 51537] {aka HSPC242, MTP18}, Glb1 (galactosidase, beta 1) [NCBI Gene 12091] {aka Bge, Bgl, Bgl-e, Bgl-s, Bgl-t, Bgs}, Drd4 (dopamine receptor D4) [NCBI Gene 13491] {aka D4R, Drd-4}, Fstl4 (follistatin-like 4) [NCBI Gene 320027] {aka B230374F23Rik, SPIG1, mKIAA1061}, Neurod1 (neurogenic differentiation 1) [NCBI Gene 18012] {aka BETA2, BHF-1, Nd1, Neurod, bHLHa3}, Foxp2 (forkhead box P2) [NCBI Gene 114142] {aka 2810043D05Rik, CAG-16, D0Kist7}, Slc18a3 (solute carrier family 18 (vesicular monoamine), member 3) [NCBI Gene 20508] {aka VAChT, VAT}, Cartpt (CART prepropeptide) [NCBI Gene 27220] {aka Cart}, Hoxd10 (homeobox D10) [NCBI Gene 15430] {aka Hox-4.5, Hox-5.3}, Mtfp1 (mitochondrial fission process 1) [NCBI Gene 67900] {aka 1700020C11Rik, 2610507A21Rik, Mtp18}, Rbpms (RNA binding protein gene with multiple splicing) [NCBI Gene 19663] {aka 2010300K22Rik, 2700019M19Rik, RBP-MS, hermes}, Pcdh9 (protocadherin 9) [NCBI Gene 211712]
- **Chemicals:** CuSO4 (MESH:D019327), sodium citrate (MESH:D000077559), Alexa Fluor 555 azide (-), Alexa Fluor 488 (MESH:C000711379), thymidine (MESH:D013936), corn oil (MESH:D003314), sucrose (MESH:D013395), PFA (MESH:C003043), 4-hydroxytamoxifen (MESH:C016601), 4-OHT (MESH:C032278), calcium (MESH:D002118), 4', 6-diamidino-2-phenylindole (MESH:C007293), Tween-20 (MESH:D011136), OCT (MESH:C051883), Triton X-100 (MESH:D017830), TM (MESH:D013629), isoflurane (MESH:D007530), ascorbic acid (MESH:D001205), 5-Ethynyl-2'-deoxyuridine (MESH:C031086), progesterone (MESH:D011374), EdU (MESH:C022811)
- **Species:** Gallus gallus (bantam, species) [taxon 9031], Mus musculus (house mouse, species) [taxon 10090]
- **Cell lines:** CD-1 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_5731), C57BL/6 — Mus musculus (Mouse), Transformed cell line (CVCL_C0MU), TM11.5 — Homo sapiens (Human), Transformed cell line (CVCL_E853), TM15.5 — Rattus norvegicus (Rat), Rat malignant glioma, Cancer cell line (CVCL_8780)

## Full text

_Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12929473/full.md

## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12929473/full.md

## References

65 references — full list in the complete paper: https://tomesphere.com/paper/PMC12929473/full.md

---
Source: https://tomesphere.com/paper/PMC12929473