Immunohistochemical localization of the ACTH (MC-2) receptor in the rat placenta and adrenal gland.

The adrenocorticotropic hormone (ACTH) acts on adrenocortical cells and promotes steroidogenesis by specific binding to the ACTH (MC-2) receptor (ACTHR). To gain an insight into ACTH action on local steroidogenic organs, we examined the immunohistochemical expression of ACTHR in rat adrenal glands and placentas during the mid-late gestation period. Antibodies against synthetic ACTHR peptides were raised in rabbits, and Western blot analysis showed that the antibody reacted with specific proteins in the rat adrenal glands and placentas.

Prevention of death of axotomized hypoglossal neurones and promotion of regeneration by chitin grafting.

1. Chitin is known to promote skin wound healing. In this study, chitin, prepared from Zuwai crab shell, was used as a bridge between the proximal and distal stumps of cut hypoglossal nerves in shrews.

Shift of hepatitis E virus RNA from hepatocytes to biliary epithelial cells during acute infection of rhesus monkey.

Hepatitis E virus (HEV) has been considered to be the major cause of enterically transmitted non-A, non-B hepatitis in developing countries. However, little is known about viral replication and localization in the liver. The aim of this study was to examine the distribution of HEV-infected cells in experimentally infected animals.

Role of apoptosis of thyrocytes in a rat model of goiter. A possible involvement of Fas system.

Apoptosis, a physiological process of cell death, may modulate the mass of the thyroid gland. We investigated the role of apoptosis and the possible involvement of Fas/Fas ligand (FasL) system in apoptosis during goiter formation and involution in a rat model of goiter. Rats were fed a low iodine diet and a goitrogen, 6-propyl-2-thiouracil, to induce goiter.

Expression of decay accelerating factor mRNA and complement C3 mRNA in human diseased kidney.

Background: Decay accelerating factor (DAF), a product of mesangial cells in vitro, is expressed on the surface of cells and is a candidate for the focal suppression of complement activation. It is not clear at present whether the levels of expression of DAF and intrarenal C3 synthesis correlate with the level of tissue injury.

Methods: Immunohistochemistry for DAF and C3 and nonradioactive in situ hybridization with digoxigenin-labeled oligonucleotide probe for DAF and C3 mRNA were performed in 22 tissue samples of kidneys from patients with IgA nephropathy (IgAN), 6 with membranous nephropathy (MN), 6 with lupus nephritis (LN), and five normal kidneys.

Activation of bullous pemphigoid antigen gene in mouse ear epidermis by ultraviolet radiation.

Bullous pemphigoid (BP) is an autoimmune blistering disease and is a photoaggravated dermatosis, but the mechanism of the aggravation is still unknown. Since damage to DNA initiates transcription of some genes, we investigated in epidermis of mouse ears the relationship between DNA damage by ultraviolet (UV) radiation and BP antigen (BP-Ag) gene activation. For this, albino male mice were irradiated with 254 nm wavelength UV for a total dose of 500 J m-2.

Autonomous cell death of mouse male germ cells during fetal and postnatal period.

Germ cell degeneration is common in mammalian testes during the developmental as well as the adult period. To investigate the extent and mechanisms of male germ cell death during fetal and neonatal life, the testes of mice at various fetal and postnatal ages extending from 13 days of gestation to 7 wk after birth were examined by electron microscopy and/or terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Electron microscopy revealed that the number of cells with typical features of spermatogenic cell apoptosis was highest at 13 days of gestation, coinciding with the time of immigration of primordial germ cells into gonads.

Cellular expression of beta-microseminoprotein (beta-MSP) mRNA and its protein in untreated prostate cancer.

Background: Previous studies have shown that beta-microseminoprotein (beta-MSP) may be used as a diagnostic marker for prostate cancer. However, the level of expression of beta-MSP in prostate cancer detected by immunohistochemistry (IHC) has varied from one study to another.

Methods: We analyzed the expression of both beta-MSP mRNA and its protein in a large sample of prostate tumors from 104 patients with untreated prostate cancer, using both nonradioactive in situ hybridization (ISH) and IHC.

Differential expression of IFN-gamma, IL-4, IL-10, and IL-1beta mRNAs in decalcified tissue sections of mouse lipopolysaccharide-induced periodontitis mandibles assessed by in situ hybridization.

To examine the role of T cell subgroups, Th1 and Th2, in the development of periodontitis, the expression of various cytokines was investigated in a mouse model of alveolar bone resorption using in situ hybridization (ISH) with digoxigenin-labeled oligonucleotides. When mice received repetitive injections with Escherichia coli lipopolysaccharide into the gingiva every 48 h, alveolar bone resorption was detectable after the fourth injection, reaching a maximum after the 13th injection. For the best performance of ISH, we first had to choose a decalcification protocol.

Induction of apoptosis in ischemia-reperfusion model of mouse kidney: possible involvement of Fas.

Although ischemia-reperfusion of mouse kidney is known to cause severe renal failure due to tubular cell death, the exact cellular mechanism responsible for this phenomenon is not clear. To investigate the spatial and temporal development of renal cell death and the role of Fas/APO-1/CD95 (Fas) in this process, the left renal vessels were occluded in a group of mice for 30, 60, or 120 min followed by reperfusion for 24 h (n = 4 for each group). Analysis of the isolated DNA in agarose-gel electrophoresis revealed a typical ladder pattern of bands consisting of multiples of 180 to 200 bp, considered the hallmark of apoptosis.

Frequency of apoptosis relates inversely to invasiveness and metastatic activity in human colorectal cancer.

The frequency of apoptosis was determined in 102 cases of human colorectal cancer. The results were correlated with the frequency of cell proliferation and with clinicopathological characteristics such as degree of differentiation, invasiveness and metastasis. As a marker of apoptosis, intranuclear DNA strand breaks were localized with in situ nick translation (ISNT).

Intraglomerular C3 synthesis in rats with passive Heymann nephritis.

Passive Heymann nephritis (PHN), a model of human membranous nephropathy, is an immune-complex-mediated glomerulonephritis characterized by the presence of complement-dependent tissue injury. Recent studies have confirmed the synthesis of C3, involved in both the classical and alternative pathways of complement, in injured human and animal renal tissues. However, there is little clear information on the role of local C3 synthesis in the pathogenesis of nephritides such as PHN.

Fas and Fas ligand interaction is necessary for human osteoblast apoptosis.

We investigated the cellular and humoral interactions between peripheral blood mononuclear cells (PBMCs) and human osteoblasts, leading to apoptosis of osteoblasts. Human osteoblastic cell line MG63 and human primary osteoblast-like cells obtained from biopsy specimens were used in this study. PBMCs were isolated from healthy donors and cultured with or without stimulation by recombinant interleukin-2 followed by 12-o-tetradecanoylphorbol 13-acetate with ionomycin.

Involvement of IL-4 in human glomerulonephritis: an in situ hybridization study of IL-4 mRNA and IL-4 receptor mRNA.

Interleukin-4 (IL-4) has been recently implicated in the pathogenesis of glomerulonephritis. However, the expression of IL-4 and IL-4 receptor (IL-4R) in human kidney has not been fully determined. Nonradioactive in situ hybridization was used to examine the expression of IL-4 mRNA and IL-4R mRNA in tissues from normal kidneys and specimens from a variety of human kidney diseases.

Expression of estrogen and progesterone receptors in endometrium and peritoneal endometriosis: an immunohistochemical and in situ hybridization study.

Objective: To clarify the role of ovarian steroids in the development and progression of endometriosis, estrogen receptors (ERs) and progesterone receptors (PRs) were localized by immunohistochemistry, and ER messenger RNA (mRNA) was detected by in situ hybridization in the uterine endometrium and in normal and altered pelvic peritoneum.

Design: Retrospective and prospective study.

Setting: Nagasaki University School of Medicine, Nagasaki, Japan.

In situ localization of basic fibroblast growth factor protein and mRNA in the retina.

Basic fibroblast growth factor (FGF) protein has been recognized as a potent factor for angiogenesis and as a mitogen. The sites of basic FGF in the mammalian retina have varied from report to report. On the other hand, only the inner segments of the photoreceptor cells have been reported to synthesized basic FGF as revealed by the presence of mRNA for basic FGF by in situ hybridization.

Intraglomerular C3 synthesis in human kidney detected by in situ hybridization.

Complements 3 and 4 are known to be synthesized in diseased renal tissue and the mRNA of these complements has been demonstrated, using polymerase chain reaction, in renal biopsies from nephritic patients. However, the types of cells producing the complements in intact renal tissue have not been defined. To identify the renal cellular components involved in complement synthesis, we analyzed the expression of C3 mRNA in renal tissues from patients with immune-complex glomerulonephritis by a high-resolution in situ hybridization using digoxigenin-labeled oligonucleotide.

Apoptosis of crypt epithelial cells in ulcerative colitis.

In the colon of ulcerative colitis (UC) patients, apoptotic bodies have been recognized in routine histopathological preparations. To investigate the extent of the apoptosis, colonic biopsies were examined from involved and uninvolved areas of untreated active UC and from normal areas in patients with colonic polyps, utilizing various markers of apoptosis. The markers included DNA breaks detected by TUNEL, Fas (CD95/APO-1) and Fas ligand (Fas-L) localized by immunohistochemistry, electron microscopic features of apoptosis, and laddering of extracted DNA.

[Expression of Fas and Fas-ligand in epithelium of ulcerative colitis].

Ulcerative colitis (UC) is characterized by the loss of epithelium and inflammation of lamina propria. In normal colon, epithelial cells are eliminated by apoptosis at the luminal surface. The apoptotic cells recognized by their typical morphology and the presence of DNA breaks are also accompanied by Fas and Fas-ligand.

[Detection of apoptosis and expression of Fas antigen in human colorectal cancer].

The occurrence of apoptosis in formalin-fixed, paraffin-embedded human colorectal cancer tissues was investigated at a cellular level by in situ nick translation (ISNT). Then the frequency of ISNT-positive nuclei was compared with the proliferative activity assessed by proliferating cell nuclear antigen (PCNA) labeling index, and with the incidence of Fas positive cells examined immunohistochemically with the incidence of Fas positive cells examined immunohistochemically with rabbit anti-Fas serum. As a result, although no significant correlation between the frequency of apoptosis and the proliferative activity was observed, a balance between them may explain a variety of growth rates of colorectal cancers.

Fas/APO-1/CD95 system as a mediator of granulosa cell apoptosis in ovarian follicle atresia.

Current studies have shown that atresia of ovarian follicles is induced through apoptosis in granulosa cells. Several articles have been devoted to study of the molecular mechanisms responsible for APO-1/CD95 (Fas) is a cell surface protein that can mediate apoptosis in lymphoid cells, and Fas ligand was recently identified in a cytotoxic T cell line. To clarify the involvement of the Fas-Fas ligand system in granulosa cell apoptosis, we investigated the expression of Fas and Fas ligand at an individual cell level.

Recent advances in molecular histochemical techniques: in situ hybridization and southwestern histochemistry.

Over the past decade, considerable efforts to understand the states of specific gene expression at cellular and/or subcellular levels have been made. For this particular purpose, nonradioactive in situ hybridization to localize mRNAs has been developed and improved substantially, and it is now recognized as a powerful, established light-microscopical technique. In this review, we focus on the recent advances in the technological aspects of nonradioactive in situ hybridization including the use of synthetic oligodeoxynucleotide probes, the progress in analysis of signals, and the application to electron microscopy.

Detection of cytokine mRNA-expressing cells in peripheral blood of patients with IgA nephropathy using non-radioactive in situ hybridization.

IgA nephropathy (IgA-N) is considered to be an immune-mediated disorder and several immunological abnormalities have been observed. In the present study, we optimized non-radioactive in situ hybridization and applied this technique to evaluate the degree of expression of various cytokine mRNAs in peripheral blood mononuclear cells (PBMC) taken from patients with IgA-N on cytospin preparation. Using this method, together with image analysis, we examined the expression of mRNA in cells which secrete cytokines, such as IL-2, interferon-gamma (IFN-gamma), IL-4, IL-5 and IL-6.

Expression of basic fibroblast growth factor and fibroblast growth factor receptor in advanced gastric carcinoma.

The expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor (FGFR) mRNA was examined in gastric carcinomas by immunohistochemistry and in situ hybridization, respectively. In the 20 advanced carcinomas examined, bFGF was found in 14 (70.0 per cent) and was confined to the tumour cells, whereas FGFR mRNA was demonstrated in 12 (60.

Expression of estrogen receptor in diseased human prostate assessed by non-radioactive in situ hybridization and immunohistochemistry.

To understand the role of estrogen in the pathogenesis of benign prostatic hyperplasia, expressions of estrogen receptor (ER) mRNA and ER protein by in situ hybridization and by immunohistochemistry, respectively, were investigated in human prostatic tissues. In non-malignant region, ER mRNA and ER protein were found in cytoplasm and nucleus, respectively, of stromal cells, but not in glandular epithelial and basal cells. In benign regions, ER mRNA/ER protein positive cells were found in fibromyoadenomatous and myoadenomatous hyperplasia, but not in adenomatous hyperplasia.