ANF mRNA is detected by the polymerase chain reaction technique in the chorioidea and bodies but not in the retina of the rat eye.

Specific proANF mRNA was demonstrated by the polymerase chain reaction (PCR) technique in ciliary body and chorioidea tissue extracts but not in the retina of the rat eye. However, immunoreactive (IR) ANF was detected in all of these tissues by a specific and sensitive radioimmunoassay (RIA). Since ANF seems to be involved in the maintenance of intraocular pressure, the regulation of ANF gene expression in these distinct eye tissues could be an important factor in the pathogenesis of glaucoma.

The effect of hypothalamic deafferentation on somatostatin-like activity in the rat brain.

Somatostatin-like activity (SLA) was determined by radioimmunoassay in several brain regions of animals which had either undergone complete deafferentation of the medial basal hypothalamus or a sham-operation. Surgical isolation of the medial basal hypothalamus caused this region to lose most of its SLA, but did not affect activity elsewhere. Similarly, partial (frontal) deafferentation of the medial basal hypothalamus resulted in a substantial decrease in SLA there.

Influence of streptozotocin-induced diabetes on the concentration of immunoreactive somatostatin in the retina and peripheral blood of the rat: effect of insulin treatment.

Changes in somatostatin-like immunoreactivity (SLI) were examined in the retina and peripheral blood of diabetic rats treated with streptozotocin (STZ) and insulin. There was no change in retinal SLI content at 4 and 11 days after administration of STZ but, thereafter, SLI increased progressively in the diabetic animals by 220% at 18 days and 300% at 27 days. Plasma SLI levels increased by 500% at 11 days and maintained similar levels thereafter. Diabetic animals treated with insulin (3-5 i.u. daily) for 27 days showed a significant (P less than 0.01) decrease of retinal and plasma SLI levels compared with untreated diabetic animals. It is concluded that there is a significant increase of retinal and plasma SLI levels in diabetic rats which tends to normalize after several days of insulin treatment.

High plasma levels of endothelin-1 and atrial natriuretic peptide in patients with acute ischemic stroke.

The plasma levels of endothelin-1 (ET-1) and atrial natriuretic peptide (ANP) have been measured in 37 patients with acute ischemic stroke, on admission and 3 and 7 days thereafter. The plasma ET-1 levels at the onset of symptoms were about two-fold those observed in age-matched normal volunteers (3.5 +/- 2.26 v 1.54 +/- 0.9 pg/mL, respectively; P < .001). These levels remained significantly elevated during the 7-day study period. The neurologic deficit was assessed daily by Mathew's modified scale (MS). A significant correlation was found between neurologic status on admission and ET-1 plasma values; patients with worse neurologic status (MS < 45 points) had higher ET-1 plasma values than those with better neurologic status (MS > 45 points) (5.4 +/- 2.34 v 3.05 +/- 2.04 pg/mL, respectively, P < .05). The plasma ET-1 values did not correlate either with the site of the infarction or with its primary cause (cardioembolic, lacunar, or atherothrombotic). No significant differences were seen in plasma ET-1 concentrations between patients who eventually died and those who survived the acute event. The plasma ANP were about 18-fold higher in ischemic stroke patients on admission than in controls at admission (110.9 +/- 29.5 v 5.84 +/- 3.96 pg/mL, respectively, P < .01). These values remained significantly elevated on days 3 and 7. There was no correlation between the ANP plasma values and the neurologic status, the site or mechanism of the stroke, or the plasma ET-1 levels. In conclusion, ischemic stroke is associated with marked acute and long-duration increases of ET-1 and ANP.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification of endothelin receptor subtypes in rat retina using subtype-selective ligands.

We investigate the existence of endothelin receptor subtypes using subtype selective ligands and the presence of immunoreactive (IR) endothelin (ET)-3 (IR-ET-3) by radioimmunoassay (RIA) in rat retina. Scatchard transformation of saturation binding experiments with [125I]ET-3 revealed specific binding sites with a Kd and Bmax values of 42 +/- 12 pM and 111 +/- 24 fmol/mg of protein, respectively. The Kd was similar to that obtain in previous studies using [125I]ET-1. However, the Bmax was 65% of that obtained with [125I]ET-1. Competitive experiments in the presence of the cyclic pentapeptide BQ123 (selective for ETA receptor) and Sarafotoxin 6C (selective for ETB receptor), demonstrated the existence of ETA and ETB receptors in a ratio of 35:65. The order of potency of ET family peptides was ET-3 = ET-1 > S6C for ETB receptor and ET-1 > ET-3 > BQ123 for ETA receptor. Cross-linking of [125I]ET-1 to retinal membranes with disuccinimidyl suberate and SDS-PAGE followed by autoradiography resulted in the labeling of two bands with apparent molecular masses of 52 and 34 kDa. Similar results were obtained using [125I]ET-3, suggesting that ETA and ETB receptors have similar molecular mass. The 34 kDa band is a proteolytic degradation product of the 52 kDa band. The concentration of IR-ET-3 was 1212 +/- 153 fmol/g wet weight in rat retina. All these data suggest that ETs may play a role in neurotransmission or neuromodulation in the retina, operating on both ETA and ETB receptor subtypes present in this tissue.

Localisation of endothelin-1 mRNA expression and immunoreactivity in the retina and optic nerve from human and porcine eye. Evidence for endothelin-1 expression in astrocytes.

We have investigated the localisation of endothelin-1 (ET-1) mRNA and ET-1-like immunoreactivity in retina and anterior portion of optic nerve from human and porcine eyes. In situ hybridisation method revealed expression of ET-1 mRNA mainly in the innermost layers of the retinas, in the retinal pigment epithelium cells as well as in the astrocytes of the optic nerve. Immunohistochemical studies showed that ET-1-like immunoreactivity appeared in the same regions where ET-1 mRNA was expressed as well as in the inner nuclear layer and in the inner segments of photoreceptors. In the nerve fibre and ganglion cell layers, astrocytes expressed both glial fibrillary acidic protein and ET-1 proteins suggesting that these cells may secrete ET-1. Expression of ETA and ETB receptors in human retina were demonstrated by reverse transcription-polymerase chain reaction. Our results demonstrated expression of ET-1 in glial, neural and vascular components of retina and optic nerve from human and porcine eyes.

Type B and type C natriuretic peptide receptors modulate intraocular pressure in the rabbit eye.

We investigated (1) the in vivo functional significance of the type B (ANP(B)) and type C (ANP(C)) natriuretic peptide receptors in the rabbit eye by evaluating the effect of intracameral administration of C-type natriuretic peptide (CNP) and C-ANP-(4-23) on intraocular pressure, and (2) the action of CNP on guanylate cyclase activity in the rabbit ciliary process membranes. Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were also studied for comparison. We demonstrated that the natriuretic peptides decrease intraocular pressure and stimulate guanylate cyclase activity, CNP being the most potent. The duration of the effect of C-ANP-(4-23) on intraocular pressure reduction was almost 9-fold that of the BNP and 20-fold that of ANP and CNP effect. This ligand increased threefold the immunoreactive natriuretic peptides levels in aqueous humour. Our data demonstrate the presence of functional ANP(A) and ANP(B) receptors in the rabbit eye and that the ANP(C) receptor modulates the concentration of the natriuretic peptides in the aqueous humour.

Higher proportions of type C than of types A and B natriuretic peptide receptors exist in the rat ciliary body.

We investigate the interaction of atrial natriuretic peptide (ANP) brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) with their receptors (NPRA, NPRB and NPRC), as well as the proportion and localization of those receptors in the rat ciliary body. Binding assays and affinity cross-linking experiments demonstrated the presence of the NPRC receptor type. However, the three natriuretic peptides stimulate the guanylate cyclase activity in the ciliary body membranes suggesting the presence of the NPRA and NPRB receptor type. Microautoradiographic data show that the NPRs are localized in the whole ciliary body. Our results indicated that NPRC is the most prominent receptor type in this tissue.

Immunoreactive atrial natriuretic factor in aqueous humor: its concentration is increased with high intraocular pressure in rabbit eyes.

Atrial natriuretic factor (ANF) concentration in the aqueous humor (AH) was studied in rabbits with experimental glaucoma induced by injecting alpha-chymotrypsin into the posterior chamber. In normal rabbit eyes, the ANF concentration in AH was 3.1 +/- 1.2 pg/ml (mean +/- SEM; n = 12), ranging from 0 to 5.8 pg/ml, whereas it was significantly higher in AH from glaucomatous rabbit eyes, being 81.0 +/- 9.8 pg/ml (n = 12). These findings were correlated with intraocular pressure (IOP), which was 13.0 +/- 2.4 mmHg (n = 12) in normal rabbit eyes and significantly greater in glaucomatous eyes: 24.4 +/- 3.0 mmHg (n = 12). Our data indicate that enhanced ANF release in AH during experimental glaucoma may play an important physiological role in modulating IOP.

Identification and characterization of atrial natriuretic factor receptors in the rat retina.

The characteristics of atrial natriuretic factor (ANF) receptors where studied in rat retinal particulate preparations. Specific 125I-ANF binding to retinal particulate preparations was greater than 90% of total binding and saturable at a density (Bmax) of 40 +/- 8 fmol/mg protein with an apparent dissociation constant (Kd) of 6.0 +/- 2.0 pM (n = 3). Apparent equilibrium conditions were established within 30 min. The Kd value of 125I-ANF binding calculated by kinetic analysis was 4.0 pM. The Bmax of 60 +/- 10 fmol/mg protein and the Kd of 5 +/- 2 pM, calculated by competition analysis, were in close agreement with the values obtained from Scatchard plots or kinetic analysis. The 125I-ANF binding to retinal particulate preparations was not inhibited by 1 microM concentration of somatostatin, vasopressin, vasoactive intestinal peptide, adrenocorticotropin, thyrotropin releasing hormone, or leu-enkephalin. The rank order of potency of the unlabelled atrial natriuretic peptides for competing with specific 125I-ANF (101-126) binding sites was rANF (92-126) greater than rANF (101-126) greater than rANF (99-126) greater than rANF (103-126) greater than Tyro-Atriopeptin I greater than hANF (105-126) greater than rANF (1-126). Similar results have been obtained in peripheral tissues and mammalian brain, indicating that central and peripheral ANF-binding sites have somewhat similar structural requirements. Affinity cross-linking of 125I-ANF to retinal particulate preparations resulted in the labelling of two sites of molecular weight 140 and 66 kDa, respectively. This demonstration of specific high-affinity ANF receptors suggests that the peptide may act as a neurotransmitter or neuromodulator in the retina.

Modelling postoperative visual acuity with and without proliferative vitreoretinopathy associated with primary rhegmatogenous retinal detachment.

PURPOSE: To find models that will explain the variability in postoperative visual acuity (VA) (logarithmic: logMAR) associated with unilateral primary rhegmatogenous retinal detachment (RD). METHODS: This was a prospective clinical cohort study of 33 patients with proliferative vitreoretinopathy (PVR: PVR

Retinal detachment: visual acuity and subretinal immunoreactive endothelin-1.

PURPOSE: To analyze whether subretinal (SRF) endothelin-1 (ET-1) - a vasoactive, mitogenic, and pro-apoptotic peptide - levels are related to visual acuity (VA) in rhegmatogenous retinal detachment (RD). PATIENTS AND METHODS: Sixty-six healthy patients between 42 and 70 years of age with unilateral RD, all candidates for scleral buckling surgery (PVR

Células progenitoras del endotelio: su posible potencial en la terapia celular de la retina isquémica / Endothelial progenitor cells: their possible potential in cell therapy for ischemic retina

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[Response of plasma and retinal somatostatin to insulin-induced hypoglycemia in diabetic and control rats]. / Respuesta de la somatostatina plasmática y retiniana a la hipoglucemia insulínica en ratas diabéticas y controles.

Changes in plasmatic levels and retinal content of somatostatin after insulin-induced hypoglycemia were investigated in three different groups of animals: Control group (C), Diabetic untreated group (D); and, Insulin-treated diabetic group (DI). In addition, another group of animals, not submitted to hypoglycemia, was used as control reference of retinal prehypoglycemic content of somatostatin (group B). Plasmatic basal levels of somatostatin were slightly higher in group DI, and significantly higher in group C, whereas they did not show any differences in group D and DI after hypoglycemia, being significantly higher in group C. The somatostatin retinal content is similar in animals not subjected to hypoglycemia and in the C and DI groups after hypoglycemia, where the rats of the D groups showed significantly higher values than the remainder of the experimental groups, an effect that is also evident in nontreated diabetic animals, even if they are not subjected to hypoglycemia, Summing up, the plasmatic somatostatin response to insulin-induced hypoglycemia is impaired in diabetic rats. Retinal somatostatin content is unchanged after hypoglycemia.

Messenger RNAs encoding the natriuretic peptides and their receptors are expressed in the eye.

The rates of secretion and removal of aqueous humour are major determinants of intraocular pressure (IOP). The natriuretic peptides, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) are vasodilators with variable effects on electrolyte and water transport at sites such as the nephron. There is some evidence that they may also affect fluid balance in the eye. As a first step in understanding the function of these peptides in the eye, we have used the technique of cDNA amplification with the polymerase chain reaction to demonstrate the presence of mRNA transcripts encoding the three natriuretic peptide receptors (NPR-A, NPR-B and NPR-C) in the retina, choroid and ciliary process of the rat and rabbit eye. In addition we have observed a differential distribution of ANP, BNP and CNP mRNAs in ocular tissues suggesting that at least part of the natriuretic peptide immunoreactivity detected in the eye arises from local synthesis of peptide. Thus, the eye appears to be able to synthesize all the components of the natriuretic peptide system necessary to modulate IOP independently of changes in the plasma concentrations of these peptides.

Identification of glucagon receptors in rat retina.

In this study, we characterize the glucagon receptors on rat retinal particulate preparations. The specific binding of 125I-glucagon was saturable and reversible. Apparent equilibrium conditions were established within 30-45 min. Analysis of binding data is compatible with the existence of two classes of binding sites: a high-affinity class with a KD of 7 +/- 0.8 nM and a Bmax of 2.3 +/- 0.2 pmol/mg of protein and a low-affinity class with a KD of 84.4 +/- 2.5 nM and a Bmax of 16.5 +/- 2.3 pmol/mg of protein. The 125I-glucagon binding to retinal particulate preparation was not inhibited by 1 microM concentrations of insulin, atrial natriuretic factor, angiotensin II, somatostatin, and vasoactive intestinal peptide. However, synthetic human pancreatic growth hormone-releasing factor, hGRF-44, inhibited binding, although the concentration required for half-maximal displacement was 10-fold higher than that for native glucagon. Glucagon binding was GTP sensitive. Inclusion of 0.1 mM GTP in the binding assay produced an increase in the concentration of unlabeled glucagon required for half-maximal displacement of 125I-glucagon, from 23 to 220 nM. Glucagon stimulated adenylate cyclase formation in retinal particulate preparations. The concentration of glucagon required for half-maximal activation of retinal adenylate cyclase was 16.2 nM. These results suggest that glucagon may play a role as a neurosignal transmitter in rat retina.