Francisco M Nadal-Nicolás1, Maria H Madeira2, Manuel Salinas-Navarro1, Manuel Jiménez-López1, Caridad Galindo-Romero1, Arturo Ortín-Martínez1, Ana Raquel Santiago3, Manuel Vidal-Sanz1, Marta Agudo-Barriuso1. 1. Instituto Murciano de Investigación Biosanitaria-VIRGEN DE LA ARRIXACA (IMIB-ARRIXACA), Murcia, Spain 2Departamento de Oftalmología, Facultad de Medicina, Universidad de Murcia, Murcia, Spain. 2. Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, University of Coimbra, Coimbra, Portugal. 3. Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, University of Coimbra, Coimbra, Portugal 4Center for Neuroscience and Cell Biology-IBILI, University of Coimbra, Coimbra, Portugal 5Association for Innovation and Biomedical.
Abstract
PURPOSE: To investigate the effect of retrograde tracing or axotomy on melanopsin mRNA expression and immunodetection in albino and pigmented rat retinas. METHODS: Groups were (1) intact-naïve retinas; (2) optic nerve crush (ONC) analyzed at 7 days (7d) or 2 months (2m); (3) Fluorogold (FG) tracing from the superior colliculi (SCi) analyzed at 7d or 2m; (4) tracing from the intact optic nerve (ON) with FG or hydroxystilbamidine methanesulfonate (OHSt), analyzed 3d later; and (5) sham tracing from the ON or sham surgery. Brn3a and melanopsin were double stained in whole mounts to quantify and assess the distribution of orthotopic and displaced Brn3a(+) retinal ganglion cells (Brn3a(+)RGCs) and melanopsin(+)RGCs (m(+)RGCs). Freshly dissected retinas were used for melanopsin mRNA quantitative PCR. RESULTS: Tracing from the SCi did not affect the number of Brn3a(+)RGCs or m(+)RGCs counted in pigmented rats. However, only 55% of m(+)RGCs were immunodetected in albinos at 7d, although by 2m the m(+)RGCs counts returned to normal. Optic nerve tracing had a more dramatic effect (38% or 77% of m(+)RGCs were immunodetected in albino or pigmented rats) that occurred irrespectively of the tracer (OHSt or FG). This effect was not observed in the sham groups. After ONC, Brn3a(+)RGCs decreased to 37% and 8% by 7d and 2m, respectively. Melanopsin (+)RGC counts diminished to 30% at 7d, but recovered to 49% of controls by 2m. Melanopsin mRNA was downregulated after ON tracing or 7d after ONC, but did not differ from intact values 2m after ONC. CONCLUSIONS: Following ON injury or retrograde tracing there is a transient melanopsin downregulation that should be taken into account when assessing m(+)RGC survival.
PURPOSE: To investigate the effect of retrograde tracing or axotomy on melanopsin mRNA expression and immunodetection in albino and pigmented rat retinas. METHODS: Groups were (1) intact-naïve retinas; (2) optic nerve crush (ONC) analyzed at 7 days (7d) or 2 months (2m); (3) Fluorogold (FG) tracing from the superior colliculi (SCi) analyzed at 7d or 2m; (4) tracing from the intact optic nerve (ON) with FG or hydroxystilbamidine methanesulfonate (OHSt), analyzed 3d later; and (5) sham tracing from the ON or sham surgery. Brn3a and melanopsin were double stained in whole mounts to quantify and assess the distribution of orthotopic and displaced Brn3a(+) retinal ganglion cells (Brn3a(+)RGCs) and melanopsin(+)RGCs (m(+)RGCs). Freshly dissected retinas were used for melanopsin mRNA quantitative PCR. RESULTS: Tracing from the SCi did not affect the number of Brn3a(+)RGCs or m(+)RGCs counted in pigmented rats. However, only 55% of m(+)RGCs were immunodetected in albinos at 7d, although by 2m the m(+)RGCs counts returned to normal. Optic nerve tracing had a more dramatic effect (38% or 77% of m(+)RGCs were immunodetected in albino or pigmented rats) that occurred irrespectively of the tracer (OHSt or FG). This effect was not observed in the sham groups. After ONC, Brn3a(+)RGCs decreased to 37% and 8% by 7d and 2m, respectively. Melanopsin (+)RGC counts diminished to 30% at 7d, but recovered to 49% of controls by 2m. Melanopsin mRNA was downregulated after ON tracing or 7d after ONC, but did not differ from intact values 2m after ONC. CONCLUSIONS:Following ON injury or retrograde tracing there is a transient melanopsin downregulation that should be taken into account when assessing m(+)RGC survival.
Authors: Eric R Bray; Benjamin J Yungher; Konstantin Levay; Marcio Ribeiro; Gennady Dvoryanchikov; Ana C Ayupe; Kinjal Thakor; Victoria Marks; Michael Randolph; Matt C Danzi; Tiffany M Schmidt; Nirupa Chaudhari; Vance P Lemmon; Samer Hattar; Kevin K Park Journal: Neuron Date: 2019-06-26 Impact factor: 17.173
Authors: Francisco M Nadal-Nicolás; Manuel Jiménez-López; Manuel Salinas-Navarro; Paloma Sobrado-Calvo; Manuel Vidal-Sanz; Marta Agudo-Barriuso Journal: J Neuroinflammation Date: 2017-11-09 Impact factor: 8.322
Authors: Manuel Vidal-Sanz; Caridad Galindo-Romero; Francisco J Valiente-Soriano; Francisco M Nadal-Nicolás; Arturo Ortin-Martinez; Giuseppe Rovere; Manuel Salinas-Navarro; Fernando Lucas-Ruiz; Maria C Sanchez-Migallon; Paloma Sobrado-Calvo; Marcelino Aviles-Trigueros; María P Villegas-Pérez; Marta Agudo-Barriuso Journal: Front Neurosci Date: 2017-04-26 Impact factor: 4.677
Authors: Marta Agudo-Barriuso; Francisco M Nadal-Nicolás; María H Madeira; Giuseppe Rovere; Beatriz Vidal-Villegas; Manuel Vidal-Sanz Journal: Neural Regen Res Date: 2016-08 Impact factor: 5.135