Systemic treatment with 7,8-Dihydroxiflavone activates TtkB and affords protection of two different retinal ganglion cell populations against axotomy in adult rats

Exp Eye Res. 2021 Sep:210:108694. doi: 10.1016/j.exer.2021.108694. Epub 2021 Jul 8.

Abstract

Purpose: To analyze responses of different RGC populations to left intraorbital optic nerve transection (IONT) and intraperitoneal (i.p.) treatment with 7,8-Dihydroxyflavone (DHF), a potent selective TrkB agonist.

Methods: Adult albino Sprague-Dawley rats received, following IONT, daily i.p. injections of vehicle (1%DMSO in 0.9%NaCl) or DHF. Group-1 (n = 58) assessed at 7days (d) the optimal DHF amount (1-25 mg/kg). Group-2, using freshly dissected naïve or treated retinas (n = 28), investigated if DHF treatment was associated with TrkB activation using Western-blotting at 1, 3 or 7d. Group-3 (n = 98) explored persistence of protection and was analyzed at survival intervals from 7 to 60d after IONT. Groups 2-3 received daily i.p. vehicle or DHF (5 mg/kg). Retinal wholemounts were immunolabelled for Brn3a and melanopsin to identify Brn3a+RGCs and m+RGCs, respectively.

Results: Optimal neuroprotection was achieved with 5 mg/kg DHF and resulted in TrkB phosphorylation. The percentage of surviving Brn3a+RGCs in vehicle treated rats was 60, 28, 18, 13, 12 or 8% of the original value at 7, 10, 14, 21, 30 or 60d, respectively, while in DHF treated retinas was 94, 70, 64, 17, 10 or 9% at the same time intervals. The percentages of m+RGCs diminished by 7d-13%, and recovered by 14d-38% in vehicle-treated and to 48% in DHF-treated retinas, without further variations.

Conclusions: DHF neuroprotects Brn3a + RGCs and m + RGCs; its protective effects for Brn3a+RGCs are maximal at 7 days but still significant at 21d, whereas for m+RGCs neuroprotection was significant at 14d and permanent.

Keywords: 7,8-Dihydroxiflavone; Adult albino rats; Apoptosis; Axotomy; BDNF neuroprotection; BDNF-Mimetic; Brn3a; Intraorbital optic nerve transection; Intrinsically photosensitive retinal ganglion cells; Melanopsin; Neuroprotection; Optic nerve section; Retinal ganglion cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axotomy
  • Blotting, Western
  • Cell Survival / physiology
  • Female
  • Flavones / administration & dosage*
  • Immunohistochemistry
  • Injections, Intraperitoneal
  • Neuroprotection
  • Neuroprotective Agents / administration & dosage*
  • Optic Nerve / physiopathology
  • Optic Nerve / surgery
  • Phosphorylation
  • Rats
  • Rats, Sprague-Dawley
  • Receptor, trkB / metabolism*
  • Retinal Ganglion Cells / drug effects*
  • Retinal Ganglion Cells / metabolism
  • Retinal Ganglion Cells / pathology
  • Rod Opsins / metabolism
  • Transcription Factor Brn-3A / metabolism

Substances

  • 6,7-dihydroxyflavone
  • Flavones
  • Neuroprotective Agents
  • Rod Opsins
  • Transcription Factor Brn-3A
  • melanopsin
  • Ntrk2 protein, rat
  • Receptor, trkB