Galvan MA, Shramm PA, Bouajram R, Lappi DA, Ancheta LR (2019) A high efficacy selection method for transfected cells utilizing recombinant isolectin B4-saporin. Neuroscience 2019 Abstracts 794.10. Society for Neuroscience, Chicago, IL.

Summary: Transfection protocols often rely on the use of antibiotics for the selection of transfected cells and has become the accepted approach for in vitro research and therapeutic applications. Antibiotics have several shortcomings such as cost, continuous use, and harmful effects — even on the transfected cell population. In addition, selection pressures are often inefficient and fail to provide a population of cells that express the gene of interest (GOI) at high levels. We have used three separate GOI’s to select for solely high-expressing transfectants using targeted toxin selection pressure. Normal Rat Kidney Cells (KNRK) were individually transfected to express green fluorescent protein (GFP), melanopsin or the low-affinity nerve growth factor receptor (p75) using an innovative new transfection delivery vector called pGEI. The results from various assays were utilized to visually determine the expression rate and pattern of the targeted toxin selection method. Melanopsin and p75 — a photopigment and nerve growth factor, respectively — were of great interest to express in our transfected cells as a means to study their role in the development and function of neurons. The delivery vector, pGEI, removes resident Galalpha(1-3)Gal epitopes from non- human mammalian cell surfaces. This residue is the target of recombinant Isolectin B4 – Saporin (IB4-SAP), a selective targeted toxin. IB4-SAP is extremely potent, with an EC50 in the low picomolar range for alpha-D-galactopyranoside expressing cells in vitro. The cells with the highest expression of the inserted vector, and therefore the GOI, will have these residues removed. Those that fail to express the vector or do not express the vector in high enough amounts, will not have all the residues removed, and will be targeted and eliminated via IB4-SAP. This method of selection provides a means of purifying the highest- expressing transfected populations using a more cost-effective and time-saving approach.

Related Products: IB4-SAP (Cat. #IT-10)

Verweij J, Chaney SY, Bredl D, Vemaraju S, König GM, Kostenis E, Lang RA, Copenhagen DR (2019) Light responses of melanopsin-expressing ganglion cells in the foetal mammalian retina. bioRxiv 675702. doi: 10.1101/675702

Related Products: Melanopsin Rabbit Polyclonal, affinity-purified (Cat. #AB-N39)

Alarautalahti V, Ragauskas S, Hakkarainen JJ, Uusitalo-Järvinen H, Uusitalo H, Hyttinen J, Kalesnykas G, Nymark S (2019) Viability of mouse retinal explant cultures assessed by preservation of functionality and morphology. Invest Ophthalmol Vis Sci 60(6):1914-1927. doi: 10.1167/iovs.18-25156 PMID: 31042799

Summary: Organotypic retinal explant cultures have been used to study retinal development, retinal diseases and injuries, drug screening, and retinal stem cell therapies.

Usage: The amount of ganglion cells and melanopsin-expressing intrinsically photosensitive RGCs (ipRGCs) were detected by staining of RNA-binding protein. Melanopsin detection by immunostaining. 1:2500

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Lin M-S, Liao P-Y, Chen H-M, Chang C-P, Chen S-K, Chern Y (2019) Degeneration of ipRGCs in mouse models of huntington’s disease disrupts non-image-forming behaviors before motor impairment. J Neurosci 39(8):1505. doi: 10.1523/JNEUROSCI.0571-18.2018 PMID: 30587542

Summary: Results show that M1 ipRGCs were susceptible to the toxicity caused by mutant Huntingtin. The resultant impairment of M1 ipRGCs contributed to the early degeneration of the ipRGC–SCN pathway and disrupted circadian regulation during HD progression.

Usage: Immunostaining (1:3000)

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Quattrochi LE, Stabio ME, Kim I, Ilardi MC, Michelle Fogerson P, Leyrer ML, Berson DM (2019) The M6 cell: A small-field bistratified photosensitive retinal ganglion cell. J Comp Neurol 527(1):297-311. doi: 10.1002/cne.24556 PMID: 30311650

Summary: M6 cells express low levels of melanopsin and have correspondingly weak intrinsic light responses.

Usage: In all experiments involving melanopsin immunofluorescence, melanopsin immunodetection was enhanced using tyramide signal amplification coupled with horseradish peroxidase (HRP)-paired with goat anti-rabbit IgG and an Alexa fluorophore. 1:10,000.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Sweeney NT, James KN, Nistorica A, Lorig-Roach RM, Feldheim DA (2019) Expression of transcription factors divides retinal ganglion cells into distinct classes. J Comp Neurol 527(1):225-235. doi: 10.1002/cne.24172 PMID: 28078709

Usage: Melanopsin (Opn4) Immunostaining 1:1000

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Dhande OS, Seabrook TA, Phan AH, Salaly LD, Ishiko N, Nguyen PL, Wang JT, Evans SM, Huberman AD (2018) Assembly of functionally antagonistic visual circuits for controlling pupil dynamics. bioRxiv 500868. doi: 10.1101/500868

Objective: Investigate the mechanisms controlling the specification and establishment of parallel sensory pathways.

Summary: Identification of a novel genetic program that marks and is required for the development of non-image-forming parallel visual pathways.

Usage: Immunohistochemistry (1:1000)

Related Products: Melanopsin Rabbit Polyclonal, affinity-purified (Cat. #AB-N39)

Ostrin LA, Strang CE, Chang K, Jnawali A, Hung L-F, Arumugam B, Frishman LJ, Smith EL, Gamlin PD (2018) Immunotoxin-induced ablation of the intrinsically photosensitive retinal ganglion cells in rhesus monkeys. Front Neurol 9:1000. doi: 10.3389/fneur.2018.01000

Objective: To develop and validate a targeted ipRGC immunotoxin to ultimately examine the role of ipRGCs in macaque monkeys.

Summary: Findings demonstrated that Melanopsin-SAP was specific for ipRGCs, and induced a graded reduction in the PLR (pupillary light reflex), as well as in ipRGC-driven pupil response with concentration.

Usage: Solutions of 0.316, 1, 3.16, 10, and 50 μg delivered intravitreally.

Related Products: Melanopsin-SAP (Cat. #IT-44)

Garrett AM, Khalil A, Walton DO, Burgess RW (2018) DSCAM promotes self-avoidance in the developing mouse retina by masking the functions of cadherin superfamily members. Proc Natl Acad Sci U S A 115:E10216-E10224. doi: 10.1073/pnas.1809430115

Summary: Focus on DSCAM (Down syndrome cell adhesion molecule 1) self-avoidance function in the mouse retina. DSCAM and members of the cadherin superfamily have also emerged as key contributors to a variety of neurodevelopmental disorders, including autism, schizophrenia, bipolar disease, Down syndrome and intellectual disability.

Related Products: Melanopsin-SAP (Cat. #IT-44)

Sabbah S, Papendorp C, Koplas E, Beltoja M, Etebari C, Gunesch AN, Carrete L, Kim MT, Manoff G, Bhatia-Lin A, Zhao T, Schreck D, Dowling H, Briggman KL, Berson DM (2018) Synaptic circuits for irradiance coding by intrinsically photosensitive retinal ganglion cells. bioRxiv 442954. doi: 10.1101/442954

Objective: To explore the synaptic networks responsible for the unique capacity of intrinsically photosensitive retinal ganglion cells (ipRGCs) to encode overall light intensity. This luminance signal is crucial for circadian, pupillary and related reflexive responses light.

Summary: Most ipRGCs sample from all bipolar terminals costratifying with their dendrites, but M1 cells avoid all OFF bipolar input and accept only ectopic ribbon synapses from ON cone bipolar axonal shafts. These monad synapses are equipped with as many as a dozen ribbons and only one postsynaptic process.

Usage: Immunohistochemistry of retina – after recording, retinas were fixed and counterstained with rabbit anti-melanopsin (1:1000) to enhance the fluorescence of the GFP-based GCaMP6f indicator.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38), Metabotropic Glutamate Receptor 2 (mGluR2) Mouse Monoclonal (Cat. #AB-N32)