Integrins are heterodimeric glycoproteins that have been found to undergo dynamic temporal and spatial changes in the endometrium during the menstrual cycle and in early pregnancy. Specificity of integrins is known to be different in human endometrial stromal cells and decidual cells. These shifts of integrins suggested to play an important role in embryo implantation and can be modulated by progesterone, cAMP derivatives, and cytokines. The mechanisms of decidualization and its precise physiological role are still not clearly understood and in vitro systems could provide an alternative that overcomes limitations of studying such complex biological phenomena in vivo at the time of implantation. This study was undertaken to establish an in vitro model system for human decidualization using 8-bromo-cAMP and to investigate the characteristics of stromal integrin expression in vitro by 8-Br-cAMP. Endometrial stromal cells were isolated and cultured, and then were induced to decidualize by 0.5 mM 8-Br-cAMP for 15 days. Immunofluorescence staining and flow cytometric analyses of the integrin subunits (alpha1, alpha4, alpha5, alpha6, beta1 and alpha v beta3) were performed at day 9. In the presence of 8-Br-cAMP, the staining intensity of alpha v beta3 was significantly higher than control and measurements for alpha1, alpha4, alpha5, alpha6, and beta1 were similar. Immunofluorescent localization of the integrins reflected the differences obtained from the flow cytometric analyses described above. In summary, the expression of alpha;avbeta;b3 integrin increased in stromal cells in vitro decidualized by 8-Br-cAMP and this up-regulation of alpha v beta3 integrin expression during decidualization might influence on human implantation.
8-Bromo Cyclic Adenosine Monophosphate/*pharmacology ; Cell Size ; Cells, Cultured ; Decidua/*cytology/drug effects/metabolism ; Female ; Flow Cytometry ; Human ; Integrins/*analysis/metabolism ; Prolactin/analysis ; Stromal Cells/cytology/*drug effects/*metabolism

OBJECTIVETo study the expression of keratinocyte (KC) basement membrane (BM) related genes during the process of re-epithelialization of burn wound in scalded rats with cDNA microarray technique.

METHODSTwenty-four SD rats were inflicted with deep partial thickness scald with an area of 45 cm(2) on the back, and they were randomly divided into A [3 postscald day (PSD)], B (10 PSD), C (14 PSD) and D (re-epithelialization complete day) groups, with 6 rats in each group. Tissue samples were harvested from 1 cm of wound margin on 3, 10 and 14 PSD. On the re-epithelialization complete day, tissue samples were harvested from the center of the wound in D group and digested with enzyme into KC suspensions. Skin samples from the back of 6 uninjured rats were taken as normal control. The differential expression of KC BM related genes during different stages of re-epithelialization was assayed with cDNA microarray.

RESULTSThe expression of laminin (LN) gamma 1 (2.068) and integrin beta8 (2.200) was up-regulated on 3 PSD compared with that in control. The expression of integrin beta1 and LN receptor 1 was up-regulated on 10 and 14 PSD, (2.472 and 2.658), while that of integrin beta2 and beta1 (0.419 and 0.462) down-regulated on 10 and 14 PSD, and the expression of type IV collagen alpha1 and alpha3 was up-regulated during re-epithelialization.

CONCLUSIONThe expression of integrin beta1, LN gamma 1, LN receptor 1, type IV collagen alpha1 and alpha3 genes were up-regulated during re-epithelialization, which might be beneficial to the construction of BM in new skin and the formation of stable conjunction between KC and BM.

Animals ; Basement Membrane ; metabolism ; Burns ; metabolism ; pathology ; Collagen Type IV ; genetics ; Disease Models, Animal ; Epithelium ; metabolism ; Gene Expression ; Integrin beta1 ; genetics ; Integrins ; genetics ; Keratinocytes ; cytology ; metabolism ; Laminin ; genetics ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Regeneration ; Skin ; cytology ; Wound Healing

OBJECTIVETo search novel genes or pathways involved in the recovery process after restraint stress in rats.

METHODSWe compared the hypothalamus transcriptional profiles of two different recovery patterns (fast recovery vs slow recovery) from restraint stress in rats using oligonucleotide microarray, the recovery pattern was determined by the decrement of plasma adrenocorticotropic-hormone (ACTH) and corticosterone levels during one hour recovery period after stress. A real-time quantitative RT-PCR was applied to validate the differential expressed genes.

RESULTSAnalysis of the microarray data showed that most of genes were not differentially expressed between fast recovery group and slow recovery group. Among the differentially expressed genes we found that talin, together with serine/threonine protein phosphatase PP1-beta catalytic subunit (PP-1B) and integrin alpha-6 precursor (VLA-6) genes, were at least 1.5 fold up-regulated in the fast recovery group, while junctional adhesion molecule 1 (F11r) was 1.5 fold down-regulated in the fast recovery group.

CONCLUSIONThe results implied that integrin signaling pathway may be involved in the recovery from restraint stress in rats. The present study provided a global overview of hypothalamus transcriptional profiles during the process of recovery from the restraint stress in rats. The integrin signaling pathway seems to be involved in the recovery process, which deserves further study to clarify the integrin-mediated recovery mechanism after restraint stress.

Adrenocorticotropic Hormone ; blood ; Animals ; Corticosterone ; blood ; Disease Models, Animal ; Gene Expression Regulation ; physiology ; Integrins ; genetics ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; physiology ; Restraint, Physical ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Signal Transduction ; physiology ; Stress, Psychological ; metabolism ; physiopathology ; Time Factors