Journal: bioRxiv

Article Title: Melanopsin regulates axonal translation underlying retinohypothalamic circuit assembly

doi: 10.64898/2026.04.21.716817

Figure Lengend Snippet: (A) Experimental design showing ipRGC-TRAP approach. Retina, SCN, and dLGN tissues were collected for immunoprecipitation of HA-tagged ribosomes, followed by RNA-seq analysis in Opn4 WT, Het, and KO mice. (B) Knock-in/knock-out genetic strategy for expressing HA-tagged ribosomes in ipRGCs. (C) Three genotypes used: WT (Opn4 +/+ ::Rpl22 HA/+) lacks Cre and serve as IP controls. Het (Opn4 Cre/+ ::Rpl22 HA/+ ) expresses tagged ribosomes and retains light sensitivity. KO (Opn4 Cre/Cre ::Rpl22 HA/+ ) expresses tagged ribosomes but is light-insensitive. (D) Experimental timeline showing tissue collection at P8 (before eye-opening) and P15 (after eye-opening). (E) Confocal microscopy showing anti-HA immunolabeling in retina (top left panel) and SCN (top right panel) with CTB labeling of RHT axons overlaid (cyan, bottom right panel). (F) Differential expression analysis of P8 heterozygous ipRGC translatomes showing retina-enriched (green) and axon-enriched (blue) transcripts with labeled genes. (G) MA plot showing fold change versus expression for P8 Het ipRGC translatomes with SCN-enriched (blue, n=298) and retina-enriched (green, n=111) genes. (H) Volcano plot of P8 KO ipRGC translatomes showing retina-enriched (green) and axon-enriched (blue) transcripts with labeled genes. (I) MA plot showing fold change versus expression for P8 KO ipRGC translatomes with SCN-enriched (blue, n=409) and retina-enriched (green, n=126) genes. (J) Upset plot comparing differentially expressed genes (DEGs) across genotypes and compartments showing unique and overlapping gene sets.

Article Snippet: The Opn4-Cre transgenic mouse line (JAX stock #035925) , Ribotag mice (#029977, B6J.129(Cg) Rpl22/SjJ) and Ai9(RCL-tdT) (B6.Cg- Gt(ROSA)26Sortm9(CAG-tdTomato)Hze /J) (JAX stock #007909) were obtained from Jackson.

Techniques: Immunoprecipitation, RNA Sequencing, Knock-In, Knock-Out, Expressing, Confocal Microscopy, Immunolabeling, Labeling, Quantitative Proteomics

Journal: bioRxiv

Article Title: Melanopsin regulates axonal translation underlying retinohypothalamic circuit assembly

doi: 10.64898/2026.04.21.716817

Figure Lengend Snippet: (A) Experimental workflow showing volumetric STORM imaging approach by ultrasectioning and serial reconstruction. (B) STORM analysis was performed in N=6 biological replicates per genotype (∼50K µm 3 volume each). Quantification shows significantly reduced synapse density Opn4-KO mice (red dots) compared to Opn4-Het controls (black dots). Het mean ± S.D. (0.03 ± 0.004); KO mean ± S.D. (0.023 ± 0.002); Cohen’s d 2.5; ***p<0.001 student’s t-test. (C) Representative 3D reconstructions of individual synapses immunolabeled for VGluT2 (cyan, presynaptic marker), Bassoon (magenta, active zone protein), and Homer1/2/3 (green, postsynaptic density marker) in Opn4 Cre/+ (Het, left panels) and Opn4 Cre/Cre (KO, right panels). Scale bars = 1 μm. (D) Quantitative analysis of synaptic protein distributions showing volume (top) and intensity (bottom) measurements for VGluT2, Bassoon, and Homer1 across N=6 Het (10,814 synapses, black) and N=6 KO (10,013 synapses, red) mice. Box-and-whisker plots with violin overlays show median, quartiles, and distribution within 1.5x the interquartile range. No significant differences observed in volume or intensity measurements between genotypes. (E) Engulfment of VGluT2 (magenta) and CTB-Alexa Fluor 488 puncta (green) by microglia (grey) in the SCN. Blue puncta in 3D renderings represent colocalized VGluT2/CTB punctae. Images show maximum intensity Z-projections (left) and 3D renderings (right) of wild-type (14.31 μm Z-stack) and Opn4-KO (12.96 μm Z-stack) tissue. (F) Superplots showing VGluT2 (top) and CTB-Alexa Fluor 488 (bottom) engulfment as a percentage of microglial volume. Small circles represent measurements from individual microglia, colored by source animal. N = 7 animals per group; Wild-type: 66 cells; Opn4-KO: 69 cells. Large circles represent individual animal means, connected to their respective cells by thin lines. Box-and-whisker plots show median, quartiles, and distribution within 1.5x the interquartile range. Statistical comparison was performed using a linear mixed model with genotype as a fixed effect and animal number as a random effect.

Article Snippet: The Opn4-Cre transgenic mouse line (JAX stock #035925) , Ribotag mice (#029977, B6J.129(Cg) Rpl22/SjJ) and Ai9(RCL-tdT) (B6.Cg- Gt(ROSA)26Sortm9(CAG-tdTomato)Hze /J) (JAX stock #007909) were obtained from Jackson.

Techniques: Imaging, Immunolabeling, Marker, Whisker Assay, Comparison

Journal: bioRxiv

Article Title: Melanopsin regulates axonal translation underlying retinohypothalamic circuit assembly

doi: 10.64898/2026.04.21.716817

Figure Lengend Snippet: (A) Schematic of bulk RNA-seq analysis of retina, suprachiasmatic nucleus (SCN), and dorsal lateral geniculate nucleus (dLGN) from Opn4 wild-type (WT), Heterozygous (Opn4 Cre/+ ), and knockout mice (Opn4 Cre/Cre ) at P8. (B) Principal component analysis shows discrete transcriptome clustering by tissue type. (C) Hierarchical clustering heatmap of top differentially expressed genes between melanopsin WT, Het, and KO mice at P8 (Z-score normalized). (D) Schematic of ipRGC communication within the retina. (E) UMAP projection of Clark et al., 2019 retinal development atlas color coded by major cell class. (F) UMAP visualization of retinal cells colored by developmental age. (G) Module mapping of Opn4 WT-enriched (dark green) and Opn4 KO-enriched (light green) genes to the developmental atlas. (H) Mean module scores averaged across developmental age (E = embryonic, P = postnatal). (I) Cell type-specific enrichment analysis showing module scores for Opn4-WT and Opn4-KO differentially expressed genes across retinal cell types, with hierarchical clustering indicating cell type relationships. ****p < 0.0001, Mann-Whitney U test with Benjamini-Hochberg correction comparing mean module score between cell class and label-shuffled dataset.

Article Snippet: The Opn4-Cre transgenic mouse line (JAX stock #035925) , Ribotag mice (#029977, B6J.129(Cg) Rpl22/SjJ) and Ai9(RCL-tdT) (B6.Cg- Gt(ROSA)26Sortm9(CAG-tdTomato)Hze /J) (JAX stock #007909) were obtained from Jackson.

Techniques: RNA Sequencing, Knock-Out, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Effects of UMP, Choline, and Fish Oil on Synaptic Integrity and Motor Coordination in an Alzheimer’s Disease Mouse Model

doi: 10.3390/ijms27083342

Figure Lengend Snippet: Behavioral assessments in Alzheimer’s disease. Effects of prenatal and lactational nutritional supplementation on locomotor activity, anxiety-like behavior, motor coordination, and nociceptive response in 5XFAD mice. ( A ) Open field test: locomotor activity was expressed as the number of squares visited. ( B , C ) Elevated plus maze test: time spent in open arms ( B ) and closed arms ( C ) was measured. ( D ) Rotarod test: motor coordination was expressed as the falling latency (seconds) from an accelerating rotarod (4–40 rpm over 5 min). ( E ) Paw withdrawal test: paw withdrawal latency was measured (seconds). Data is presented as the mean ± standard error of the mean (SEM) ( n = 5–7 animals per group). Statistical significance is indicated as * p < 0.05 and ** p < 0.01.

Article Snippet: The 5XFAD transgenic mouse line [ ], which harbors five familial AD-related mutations (APP: Sweden, Florida, and London; PSEN1: M146L and L286V) on a mixed C57BL/6 × SJL genetic background (strain designation: B6SJL-Tg), was obtained from the Jackson Laboratory (Jackson Laboratory, Bar Harbor, ME, USA) (MMRRC_034840-JAX; stock no. 034848-JAX).

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Effects of UMP, Choline, and Fish Oil on Synaptic Integrity and Motor Coordination in an Alzheimer’s Disease Mouse Model

doi: 10.3390/ijms27083342

Figure Lengend Snippet: Phenotypic characterization of 5XFAD (BS6JL-Tg) mice .

Article Snippet: The 5XFAD transgenic mouse line [ ], which harbors five familial AD-related mutations (APP: Sweden, Florida, and London; PSEN1: M146L and L286V) on a mixed C57BL/6 × SJL genetic background (strain designation: B6SJL-Tg), was obtained from the Jackson Laboratory (Jackson Laboratory, Bar Harbor, ME, USA) (MMRRC_034840-JAX; stock no. 034848-JAX).

Techniques: