Rollín Raquel, Mediero Aránzazu, Roldán-Pallarés Manuela, Fernández-Cruz Arturo, Fernández-Durango Raquel
Unidad de Investigación, Departamento de Medicina Interna III, Hospital Clínico San Carlos, Madrid, Spain.
Mol Vis. 2004 Jan 9;10:15-22.
The natriuretic peptide (NP) family includes atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). Natriuretic peptides are known to inhibit vascular cell growth and regulate vessel tone. There is also much evidence to suggest they modulate vascular permeability and angiogenesis, as well as regulating aqueous humor production in the eye. All these data indicate that the natriuretic peptide system might be involved in the development of diabetic retinopathy and glaucoma. Given the expression pattern of natriuretic peptides (NPs) and their receptors, natriuretic peptide receptor A (NPRA), natriuretic peptide receptor B (NPRB) and natriuretic peptide receptor C (NPRC) in the human retina has not yet been established, the present study was designed to determine ANP, BNP and CNP gene expression and localize the mature peptides in this tissue. The expression pattern of the genes encoding the different NP receptor subtypes was also examined.
Eyes (n=10) from human donors with no history of eye disease were fixed and processed for routine paraffin embedding. The cellular location of the NPs was established by immunohistochemistry. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the expression of NP and NP receptor genes in neural retinas obtained from the contralateral eyes.
Immunohistochemistry revealed the presence of NPs in the neural retina and retinal pigment epithelium. Positive NP immunostaining was observed within the astrocytes and in their processes enveloping vessels. In the anterior portion of the optic nerve, NPs were intensely labeled in neural bundles. We were able to detect NP gene expression in the human retina. The levels of NP receptor-encoding transcripts detected indicated no significant differential expression of genes coding for the different receptor subtypes.
Our finding that NP receptor transcripts are expressed along with ANP, BNP, and CNP mRNA in the human retina provides evidence for a local system in this tissue. The expression of NPs in neural retinal, glial, and vascular elements of the normal adult retina suggests a role for these peptides in maintaining both the neural and vascular integrity of the mature retina.
利钠肽家族包括心房利钠肽(ANP)、脑利钠肽(BNP)和C型利钠肽(CNP)。已知利钠肽可抑制血管细胞生长并调节血管张力。也有大量证据表明它们可调节血管通透性和血管生成,以及调节眼内房水生成。所有这些数据表明利钠肽系统可能参与糖尿病视网膜病变和青光眼的发生发展。鉴于利钠肽(NPs)及其受体利钠肽受体A(NPRA)、利钠肽受体B(NPRB)和利钠肽受体C(NPRC)在人类视网膜中的表达模式尚未确定,本研究旨在确定ANP、BNP和CNP基因表达并在该组织中定位成熟肽。还检测了编码不同NP受体亚型的基因的表达模式。
选取10例无眼部疾病史的人类供体的眼睛进行固定,然后进行常规石蜡包埋处理。通过免疫组织化学确定NPs的细胞定位。采用实时定量逆转录-聚合酶链反应(RT-PCR)评估从对侧眼获得的神经视网膜中NP和NP受体基因的表达。
免疫组织化学显示神经视网膜和视网膜色素上皮中存在NPs。在星形胶质细胞及其包绕血管的突起内观察到阳性NP免疫染色。在视神经前部,神经束中NPs被强烈标记。我们能够在人类视网膜中检测到NP基因表达。检测到的NP受体编码转录本水平表明,编码不同受体亚型的基因无明显差异表达。
我们发现NP受体转录本与ANP、BNP和CNP mRNA一起在人类视网膜中表达,为该组织中的局部系统提供了证据。NPs在正常成人视网膜的神经视网膜、神经胶质和血管成分中的表达表明这些肽在维持成熟视网膜的神经和血管完整性方面发挥作用。