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The goals of this study were to describe the morphology, neurotransmitter content and synaptic connections of neurons in primate retinas that contain the neuropeptide secretoneurin. Amacrine cells were labeled with antibodies to secretoneurin in macaque and baboon retinas. Their processes formed three distinct plexuses in the inner plexiform layer: one in the outermost stratum, one in the center and one in the innermost stratum. In light microscopic double immunolabeling experiments, GABA was colocalized with secretoneurin in these cells, but glycine transporter 1 and Substance P were not. ON bipolar cell axon terminals labeled with antibody to the cholecystokinin precursor, G6-gly, have ON responses to stimulation of short wavelength sensitive (S) cones. Axons of these bipolar cells made contacts with amacrine cell dendrites containing secretoneurin. Secretoneurin-IR amacrine cells also made contacts with retinal ganglion cell dendrites labeled with antibody to the photopigment melanopsin, which have OFF responses to stimulation of S cones. Using electron microscopic immunolabeling, 436 synapses from macaque retina were analyzed. Axons from bipolar cells were identified by their characteristic synaptic ribbons; their synaptic densities were asymmetric like those of excitatory synapses in the brain. Amacrine cells made and received conventional synapses with symmetric synaptic densities, like those of inhibitory synapses in the brain. Ganglion cell dendrites were identified by their absence of presynaptic specializations; they received inputs from both amacrine cells and bipolar cells. The majority of inputs to the secretoneurin-IR amacrine cells were from other amacrine cells, but they also received 21% of their input from bipolar cells. They directed most of their output, 54%, to amacrine cells, but there were many synapses onto bipolar cell axons and ganglion cell dendrites, as well. The synaptic connections were very similar in the three plexuses with one notable exception; output synapses to bipolar cells were significantly less common in the innermost one, where the S-ON bipolar cells terminate. Taken together, these findings suggest that the secretoneurin-IR amacrine cells in primates receive excitatory input from S-ON bipolar cells and, in turn, inhibit intrinsically photosensitive retinal ganglion cells.

In this study, we investigated whether the proangiogenic neuropeptides secretoneurin (SN), substance P (SP), and neuropeptide Y (NPY) contribute to the development of abnormal neovascularization in the oxygen-induced retinopathy (OIR) model in mice. By exposing litters of C57Bl/6N mice to 75% oxygen from postnatal day 7 (P7) until postnatal day 11 (P11) and then returning them to normoxic conditions, retinal ischemia and subsequent neovascularization on the retinal surface were induced. Retinae were dissected on P9, P11, P12–P14, P16 and P20, and the concentrations of SN, SP, NPY and VEGF determined by radioimmunoassay or ELISA. The levels of SN and SP increased in controls from P9 until P16 and from P9 until P14, respectively, whereas the levels of NPY were high at P9 and decreased thereafter until P20, suggesting that NPY may participate in the development of the retina. However, dipeptidyl peptidase IV (DPPIV) and the NPY-Y2 receptor were not detectable in the immature retina indicating that NPY is not involved in the physiological vascularization in the retina. Compared to controls, OIR had no effect on the levels of SN, whereas levels of both SP and NPY slightly decreased during hyperoxia. Normalization of the levels of SP, and to a more pronounced extent of NPY, was significantly delayed during relative hypoxia. This clearly indicates that these three neuropeptides are not involved in the pathogenesis of neovascularization in OIR. Moreover, since there were no differences in the expression of two vessel markers in the retina of NPY knockout mice versus controls at P14, NPY is also not involved in the delayed development of the intermediate and deep vascular plexus in the retina in this animal model.

By means of highly sensitive radioimmunoassays, the levels of substance P (SP) and secretoneurin (SN) were detected in vitreous aspirates of patients with macular holes which served as controls, in patients with nonproliferative diabetic retinopathy (DR), active proliferative diabetic retinopathy (active PDR), inactive PDR, rhegmatogenous retinal detachment and proliferative vitreoretinopathy (PVR). Furthermore, SN-like immunoreactivities were characterized by reversed phase-HPLC. The concentration of SN was more than 20-fold higher in macular holes when compared with SP and reversed phase HPLC revealed evidence that the vitreous levels of SN represent authentic SN. SN was significantly decreased in patients with nonproliferative DR, active PDR and inactive PDR by more than 70% which seems to result from a reduced expression and/or secretion from the cilary epithelium and a reduced release from the retina both due to diabetes mellitus. By contrast SP was increased in rhegmatogenous retinal detachment most obviously due to an enhanced outflow of the peptide through retinal breaks. Despite their proangiogenic activities, SP and SN are unlikely to be involved in the pathogenesis of neovascularizations in DR because of their unchanged and reduced levels, respectively, but the low levels of both peptides may facilitate the regression of vasoproliferations following laser photocoagulation.

Secretoneurin (SN) is a conserved peptide derived by proteolytic processing from the middle domain of the ∼600 amino acid precursor secretogranin-II (SgII). Secretoneurin is widely distributed in secretory granules of endocrine cells and neurons and has important roles in reproduction as it stimulates luteinizing hormone release from the pituitary. A potential new role of SN in goldfish feeding is the subject of this study. Firstly, we established that acute (26 h; p

Secretoneurin (SN) was detected in the aqueous humor (AH) of patients treated topically with tobramycine eye drops alone or tobramycine and cyclosporine A, tobramycine and diclofenac or tobramycine and rimexolone. The levels of the peptide were found to be higher in the uninflamed human than in the rabbit aqueous humor which may be the result of species differences and/or age-related circumstances. Furthermore, they are approximately one hundred times higher than those of classical neuropeptides indicating release from nerve fibers and/or secretion from non-pigmented ciliary epithelium cells. Despite a slight tendency by rimexolone to decrease the levels, there was no significant effect seen for either of the drops. It must be considered that aminoglycosides are known to have toxic side effects and that they can influence the levels of SN which may be not diminished by low topical doses of corticosteroids or nonsteroidal antiinflammatory drugs. The high levels of the peptide are of relevance and may indicate a significant role of secretoneurin in the anterior segment of the eye. This should encourage performing functional studies.

Activation of neuropeptide receptors on leukocytes induces chemotaxis. We determined in Boyden chambers with micropore filters, whether in human monocytes and lymphocytes this migratory response is heparan sulfate proteoglycan (HSPG) dependent. Chemotaxis toward calcitonin gene-related peptide, secretoneurin, vasoactive intestinal peptide (VIP), and substance P (SP) was abolished by removal of heparan sulfate side chains from cell surface proteoglycans or by addition of anti-syndecan-4 antibodies. Inhibition of neuropeptide-induced chemotaxis by dimethyl sphingosine (DMS), an inhibitor of sphingosine kinase, indicates transactivation of the sphingosine-1-phosphate chemotaxis pathway which was previously identified as being syndecan-4-related. Data suggest that HSPGs are involved in neuropeptide-induced chemotaxis of leukocytes.

Chromogranins belong to an evolutionarily conserved family of proteins that serve as neuropeptide pro-proteins, besides having other functions. The secretogranin-II-derived peptide secretoneurin is a 33-amino-acid polypeptide generated by proteolytic cleavage at paired dibasic sequences that exerts its effect by binding to specific receptors. Secretoneurin receptors have been kinetically and functionally characterized indicating that they are G-protein linked. Localization of secretoneurin and functional studies have helped to elucidate roles for secretoneurin, ranging from effects in the central nervous system to the modulation of the inflammatory response in the periphery. It has been shown that secretoneurin possesses biologic activities such as stimulation of dopamine release from striatal neurons and activation of monocyte migration, suggesting that the peptide may modulate both neurotransmission and inflammatory response. With an array of actions as diverse as that seen with other sensory neuropeptides, there is scope for numerous studies and therapeutic possibilities.

Functions of secretoneurin include chemotaxis for monocytes and endothelial cells, and inhibition of endothelial cell proliferation. Inhibition of monocyte chemotaxis by staurosporine indicated involvement of specific signaling mechanisms. We have tested effects of kinase inhibitors on activation of endothelial cells for neutrophil adherence by secretoneurin. Pretreatment of endothelial cells by secretoneurin induced in endothelium increased adhesiveness to neutrophils. Addition of staurosporine, an inhibitor of protein kinase C, completely abolished endothelial cell activation, whereas tyrphostin-23, a tyrosin kinase inhibitor, had no effect. Results on activation of neutrophil-endothelial cell adherence by secretoneurin demonstrate that specific signaling mechanisms are involved in endothelial cell activation.

Secretoneurin (SN) is a neuropeptide formed by endoproteolytic processing of secretogranin II (chromogranin C). Chromatographic analysis revealed that the human retina contains significant concentrations (14.2 fmol/mg wet weight) of this peptide. Its cellular localization in the retina was characterized by immunohistochemistry. SN-immunoreactive (IR) fibers showed a distinct distribution in central and peripheral retinal regions. Immunopositive somata were found in the ganglion cell layer and in the inner nuclear layer. The localization was similar to that of substance P. The physiological role of SN in the human retina is at present unknown. However, its presence in ganglion cells and/or amacrine cells suggests that it may play a role in visual processing.