Journal: bioRxiv
Article Title: Temporally-segregated dual functions for Gfi1 in the development of retinal direction-selectivity
doi: 10.1101/2025.06.03.657700
Figure Lengend Snippet: Gfi1 is expressed in direction-selective D-oDSGC and F-mini ON RGC subsets ( A ) Schematic depicting Gfi1 expression in the retina restricted to D-oDSGCs, F-mini-ONs and W3Bs. D-oDSGCs and F-mini ONs are direction-selective subsets tuned primarily to downward motion, while W3Bs are non-direction-selective. ( B ) Wholemount P6 Gfi1 Cre , LSL-YFP retina shows YFP expression restricted to Rbpms + RGCs. ( C ) Density recovery profile (DRP) of YFP + RGCs suggests an absence of mosaicism confirming Gfi1 expression in more than one RGC subtype. ( D ) Density of Gfi1-expressing RGCs in the retina. ( E ) In situ hybridization in P4 retina cryosection shows Gfi1 expression in Fibcd1 + D-oDSGCs. ( E’, E” ) Detailed views of the nuclei indicated in D. ( F ) Quantification of the percentage of Gfi1 + RGCs co-expressing Fibcd1 . ( G ) Coronal brain section from Gfi1 Cre , LSL-tdTomato adult mouse shows strong innervation of the ventral medial terminal nucleus (MTN) by tdTomato + RGCs. ( H-N ) Detailed views of all retinorecipient nuclei labeled with cholera toxin B (CTB) from Gfi1 Cre , LSL-YFP adult mice confirms YFP + RGC innervation exclusively in the ventral MTN (H, H’ ) and the SC ( I, I’ ), and not in the other retinorecipient nuclei ( J-N ). ( O ) Wholemount P1 Gfi1 Cre , LSL-YFP retina shows YFP expression in a subset of Foxp2 + RGCs. ( P, Q ) These YFP + Foxp2 + RGCs account for ∼25% of all Foxp2 + RGCs and ∼50% of all Gfi1 + RGCs. ( R ) Combinatorial labeling can differentiate between F-RGC subsets. ( S-U ) Wholemount Gfi1 Cre , LSL-YFP retina shows that YFP + Foxp2 + RGCs do not express the F-RGC OFF marker Foxp1 or the F-midi ON marker Brn3c, confirming their F-mini ON identity. Data are presented as mean ± SE.
Article Snippet: Primary antibodies used in this study include: Rabbit anti-dsRed (Living Colors, 1:1000), Chick anti-GFP (AVES, 1:1000), Guinea Pig anti-Foxp2 (Synaptic Systems, 1:500), Rabbit anti-Rbpms (Proteintech, 1:500), Guinea Pig anti-Rbpms (ThermoFisher, 1:500), Mouse anti-Brn3c (Santa Cruz, 1:100), Rabbit anti-Foxp1 (Proteintech, 1:500), Rabbit anti-IBA1 (Wako Laboratory Chemicals, 1:500), Goat anti-ChAT (Abcam, 1:250), Goat anti-VAChT (Millipore, 1:250) and Rabbit anti-cFos (Cell Signaling Technology, 1:500).
Techniques: Expressing, In Situ Hybridization, Labeling, Marker
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