PAROTID RAB3/RAB-GDI ISOFORMS--EXPRESSION/LOCALIZATION

腮腺 RAB3/RAB-GDI 同种型——表达/定位

• 批准号: 2684015
• 负责人: ROBERT D RAFFANIELLO
• 金额: $ 2.85万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2684015
• 关键词: amylases; enzyme activity; exocytosis; gene induction /repression; guanine nucleotide binding protein; immunoelectron microscopy; laboratory rat; molecular cloning; northern blottings; nucleic acid probes; nucleic acid sequence; parotid gland; polymerase chain reaction; protein isoforms; protein structure function; secretion; western blottings

The long-term objective of the studies described in the present proposal is to elucidate the role of Rab proteins, as well as accessory proteins which regulate Rab protein function, in regulated exocytosis in the parotid gland. The specific aims of this proposal are to: 1) examine the expression and localization of Rab3 and rab-GDP dissociation inhibitor (rab-GDI) isoforms; and 2) examine agonist- induced subcellular redistribution of these isoforms in rat parotid gland. The salivary glands perform many important functions that affect the oral and gastrointestinal environment including protein secretion. Hence, an understanding of the basic mechanisms that regulate salivary gland secretion is essential. Although Rab proteins are involved in various stages of vesicular transport and secretion, the expression and function of these important proteins in salivary glands have not been studied. Four Rab3 isoforms (A, B, C and D) and at least two rab-GDI isoforms have been identified in mammalian cells. Rab3 isoform expression in the parotid gland will be examined by RT-PCR using Rab3- specific primers. The PcR products obtained will be cloned and sequenced to identify the Rab3 isoforms expressed in this tissue. Rab3 and rab-GDI expression will also be examined by Northern and Western blotting using isoform-specific DNA probes and antibodies, respectively. Preliminary studies from this laboratory indicate that at least one Rab3 isoform, Rab3D, and one rab-GDI isoform are expressed in rat parotid gland. The association of Rab3 and rab-GDI isoforms expressed in the parotid gland with various subcellular fractions, including secretory granules, will be examined by cellular fractionation and Western blotting. Precise subcellular localization of these proteins will also be studied by immunofluorescence and immunogold electron microscopy. The effects of agents which stimulate amylase secretion on the subcellular localization of Rab3 and rab-GDI isoforms will be examined by quantitative Western blotting. The studies outlined in this proposal should indicate a role for Rab3 and rab-GDI proteins in mediating regulated secretion in the parotid gland.

研究的长期目标描述在目前

项目成果

Rab3D redistribution and function in rat parotid acini.

Rab3D 在大鼠腮腺腺泡中的重新分布和功能。

• DOI: 10.1002/jcp.10373
• 发表时间: 2003
• 期刊: Journal of cellular physiology
• 影响因子: 5.6
• 作者: Nguyen,Danieele;Jones,Antoinette;Ojakian,GeorgeK;Raffaniello,RobertD;Ngyen,Danielle
• 通讯作者: Ngyen,Danielle

Expression and localization of Rab3D in rat parotid gland.

Rab3D 在大鼠腮腺中的表达和定位。

• DOI: 10.1016/s0167-4889(99)00052-x
• 发表时间: 1999
• 期刊: Biochimica et biophysica acta
• 作者: Raffaniello,RD;Lin,J;Schwimmer,R;Ojakian,GK
• 通讯作者: Ojakian,GK