Mechanical stress enhances CD9 expression in cultured podocytes.

Elevated glomerular pressure represents a high risk for the development of severe kidney diseases and causes an increase in mechanical load to podocytes. In this study, we investigated whether mechanical stress alters gene expression in cultured podocytes using gene arrays. We found that tetraspanin CD9 is significantly upregulated in cultured podocytes after mechanical stress. The differential expression of CD9 was confirmed by RT-PCR and Western blotting under stretched and unstretched conditions. Furthermore, mechanical stress resulted in a relocalization of CD9. To get an insight into the functional role of CD9, podocytes were transfected with pEGFP-CD9. The expression of CD9 induced the formation of substratum-attached thin arborized protrusions. Ca(2+) depletion revealed that podocytes overexpressing CD9 possess altered adhesive properties in contrast to the control transfected cells. Finally, elevated CD9 expression increased migration of podocytes in a wound assay. In summary, our results suggest that upregulation of CD9 may play an important role in podocyte morphology, adhesion, and migration.

[Preparation of the amniotic membrane with the waterjet]. / Zur Aufbereitung von Amnionmembranen mit dem Waterjet.

BACKGROUND: Ex vivo culturing of limbal stem cells on human amniotic membranes can be accelerated if all amniotic epithelial cells have been removed beforehand. A common application of acellular amniotic membranes is their use in cultivating autologous oral mucosal epithelial cells for transplantation in cases of bilateral stem cell insufficiency. Amniotic epithelial cells can be eliminated with enzymatic-chemical or mechanical methods or with a combination of both. MATERIAL AND METHODS: The efficacy of a waterjet cutter to eliminate amniotic epithelial cells from the amniotic membrane was investigated. Deep frozen placentas from healthy mothers were defrosted and a well-defined surface of the amniotic membrane (d = 15 mm) was treated with the waterjet in a standardized way. The waterjet used two different nozzles (pin-point and narrow stream nozzles). The applied system pressures with the pin-point stream nozzle (aperture 120 µm) were 30, 40 and 50 bar and the narrow stream nozzle was operated with pressures of 70, 80 and 90 bar on the amniotic membrane. A total number of 42 tissue samples were examined with an optical microscope using native trypan blue staining. For each type of nozzle and each application pressure two amnion samples were examined with a scanning electron microscope to analyze the efficacy of the mechanical epithelial cell elimination from the amniotic membrane. After medical imaging and histopathological examination the efficacy was graded using the following scale: 0 = no amniotic epithelial cells, 1 = no cells, low amounts of cell debris, 2 = single amniotic epithelial cells, large amounts of cell debris, 3 = loose cell layer, 4 = continuous sheet of epithelial cells. RESULTS: To eliminate epithelial cells from the surface of the amniotic membrane with the waterjet pinpoint stream nozzle (aperture: 120 µm) an application pressure of 30-50 bar was needed. The use of the narrow stream nozzle required a pressure of 70-90 bar. CONCLUSIONS: The preparation of amniotic membranes with the waterjet represents a precise option to mechanically eliminate amniotic epithelial cells from the amniotic membrane. The use of a waterjet cutter as an exclusively mechanical method without enzymatic-chemical substances may be a benefit, as cytotoxic effects on culturing limbal stem cells caused by chemical substances are not present.

The role of annexins I, II and IV in tumor development, progression and metastasis of human penile squamous cell carcinomas.

PURPOSE: The outcome of patients with penile squamous cell carcinomas (PSCC) largely depends on occurrence of metastasis. Therefore, prognostic markers indicating the risk for tumor cell spreading would be useful. Since Annexins are potential prognostic markers in a variety of tumors, we immunohistochemically examined the expression of Annexins I, II and IV (ANX AI, ANX AII and ANX AIV) in PSCC. METHODS: Samples originated from 29 patients subjected to surgical resection of invasive PSCC. Immunohistochemistry was done on paraffin-embedded sections using monoclonal antibodies against ANX AI, ANX AII and ANX AIV. Expression of ANXs was compared with clinical data. RESULTS: ANX AI expression was found in conventional PSCC and was absent in basaloid and sarcomatoid subtypes. High ANX AI score was significantly associated with higher T stages (P = 0.006). Strong expression in the invasion front of carcinomas was significantly associated with the occurrence of lymph node metastasis (P = 0.001). ANX AIV expression was weak in conventional PSCC, while it was strong in basaloid and sarcomatoid subtypes. Strong expression of Annnexin IV in the invasion front also showed a significant association with metastasis (P = 0.019). CONCLUSION: Expression of ANXs was different in histologic subtypes of penile carcinomas. Strong expression of ANX AI and ANX AIV in the invasion front seems to indicate a higher risk of lymph node metastasis.

[On the topical tolerance of aminoglycoside antibiotics--a prospective comparative cytomorphological study]. / Zur Evaluierung der lokalen Verträglichkeit von Antibiotika an der Augenoberfläche am praktischen Beispiel eines Aminoglykosides.

BACKGROUND: The local application of drugs is an efficient method for treating diseases of the external eye. However, some active substances can also cause a chronic toxic reaction at the ocular surface. No results from clinical experiments on the local tolerance of kanamycin which would meet the requirements of evidence-based medicine are as yet available. METHODS: Thus, the cytomorphological processes of the treatment with kanamycin were examined in a prospective randomised placebo-controlled double blind cross-over study. The effect of the topical application (5 times a day for 7 days) of Kanamycin POS® eye ointment on the conjunctival epithelial barrier was studied in comparison with an eye ointment base (placebo) in 25 healthy subjects. To detect cytopathological microchanges of the epithelium, we carried out a standardised impression cytology (primary compatibility parameter). Biomicroscopy, Schirmer II test and non-invasive measurement of the break-up-time (tearscope) were also examined. The subjective compatibility parameters (itching, tearing, foreign body sensation, burning sensation, mucus) were measured on a 0 - 10 visual analogue scale. RESULTS: The cytological findings of 23 subjects were included in the final evaluation of the study. The impression cytology did not reveal a different effect of kanamycin ophthalmic ointment on the conjunctival ocular surface in comparison with the placebo. Only in one subject was a marked shift found in the nuclear-cytoplasmic ratio after application of verum. No signs of increased apoptosis ("snake-like chromatin") were found at the conjunctival epithelial barrier. The mean goblet cell count was 109 goblet cells per mm (2). No significant reduction of the goblet cell density was found in any treatment group. Among the subjective compatibility parameters, only the mean for itching constantly remained above the placebo group after application of kanamycin. Statistically significant differences were not found by the Wilcoxon test. CONCLUSIONS: In contrast to the clinical experience with the systematic application of kanamycin, the standardised evaluation of this study confirmed the very good topical tolerability of the tested kanamycin eye ointment. In view of the continuing clinically relevant aspect of the availability of a wide spectrum of antibiotics to avoid the development of resistance while at the same time keeping within what is reasonable economically, Kanamycin POS® for topical application supplements the options of ophthalmological antibacterial chemotherapy.

Identification and characterization of the doublesex gene of Nasonia.

The doublesex (dsx) gene of the parasitic wasp Nasonia vitripennis is described and characterized. Differential splicing of dsx transcripts has been shown to induce somatic sexual differentiation in Diptera and Lepidoptera, but not yet in other insect orders. Two spliceforms of Nasonia dsx mRNA are differentially expressed in males and females. In addition, in a gynandromorphic line that produces haploids (normally males) with full female phenotypes, these individuals show the female spliceform, providing the first demonstration of a direct association of dsx with somatic sex differentiation in Hymenoptera. Finally, the DNA binding (DM) domain of Nasonia dsx clusters phylogenetically with dsx from other insects, and Nasonia dsx shows microsynteny with dsx of Apis, further supporting identification of the dsx orthologue in Nasonia.

Down-regulation of the metastasis suppressor protein KAI1/CD82 correlates with occurrence of metastasis, prognosis and presence of HPV DNA in human penile squamous cell carcinoma.

In penile squamous cell carcinoma (PSCC), the outcome largely depends on early detection and resection of inguinal lymph node metastases. We investigated the role of metastasis suppressor protein kang ai 1 (KAI1)/cluster of differentiation 82 (CD82), which is known to be of prognostic significance for a wide variety of cancers. Moreover, we analysed the tumours for human papillomavirus (HPV) DNA and loss of heterozygosity at the 11p11.2 locus. Tissue samples of 30 primary PSCCs were investigated immunohistochemically using an anti-KAI1/CD82 polyclonal antibody. The expression was assessed according to the degree of KAI1/CD82-positive tumour cells as positive, decreased or negative. The presence of HPV6/11, HPV16 and HPV18 DNA was analysed by polymerase chain reaction. All patients with decreased or negative expression of KAI1/CD82 in primary lesions had lymph node metastases (p = 0.0002). Patients with positive KAI1/CD82 expression showed a significant better prognosis for survival compared to the other groups (p = 0.0042). Presence of HPV DNA was associated with decreased or negative KAI1/CD82 expression. Lacking or decreased expression of metastasis suppressor gene KAI1/CD82 appears to be a prognostic parameter for the occurrence of lymph node metastases in PSCC. Our study suggests an association of decreased KAI1/CD82 expression with tumour progression, development of metastases and disease-specific death.

[Establishing a protein signature from prostate tissue biopsies]. / Erstellung eines Proteinprofils aus Gewebeproben der Prostata.

The prostate-specific antigen test (PSA) has been a major factor contributing to a better management of prostate cancer. The low specificity limits its use in diagnosis especially in early detection of prostate cancer. Multiply expressed proteins need to be identified to establish a disease-specific protein signature that distinguishes between cancerous and noncancerous tissue. The first aim of our study is to identify differentially expressed proteins in both tissues using two-dimensional gel electrophoresis and subsequent mass spectrometry. We elucidated whether prostate biopsies are useful. First results have shown a different protein expression pattern in cancerous and noncancerous tissue. PCR revealed an increasing amount of mRNA for some upregulated proteins. We conclude that biopsies are useful material to establish protein expression patterns.

Expression of proliferation marker Ki67 correlates to occurrence of metastasis and prognosis, histological subtypes and HPV DNA detection in penile carcinomas.

Clinical outcome of penile squamous cell carcinoma (PSCC) largely depends on the presence of lymph node metastasis. In search of a valuable marker predicting the risk for metastasis, the expression of Ki67 was investigated immunohistochemically in primary tumors and compared to presence of inguinal lymph node metastasis. As human papilloma virus (HPV) is thought to affect Ki67 expression, we evaluated whether occurrence of HPV DNA correlates to Ki67 score or metastatic potential. Samples originated from patients subjected to resection of invasive SCC of penis. Immunohistochemistry was done on paraffin-embedded sections using a monoclonal antibody against Ki67. After DNA isolation from paraffin embedded tissue the presence of HPV 6/11, HPV 16 and HPV 18 DNA was analyzed by PCR. Statistical analysis was done using two tail unpaired t test and Chi-square test. Four of 28 patients showed a weak Ki67 expression, without displaying lymph node metastasis. Among 17 patients showing an intermediate Ki67 index, eight exhibited metastases while in all seven patients with a strong expression of Ki67 lymph node metastases were found. The median Ki67 expression in metastastic lesions was significantly different (50.3%) from tumors without lymph node metastasis (31.8%) (p=0.024). Furthermore, a correlation between presence of HPV DNA and strong Ki67 expression was determined (p=0.009). Since our study demonstrated a strong Ki67 labeling index significantly associated to positive lymph nodes, we suggest Ki67 expression as a prognostic marker for lymph node metastasis in penile squamous carcinoma.

Expression of annexin AI in conventional renal cell carcinoma (CRCC) correlates with tumour stage, Fuhrman grade, amount of eosinophilic cells and clinical outcome.

There is increasing evidence that Annexin AI (ANX AI) expression is dysregulated in several carcinomas and tumour cell lines. In order to gain insight into the putative role of ANX AI in tumorigenesis, clinical outcome and metastatic potential of conventional renal cell carcinomas (CRCCs) we investigated the expression of ANX AI in CRCCs and metastases. Furthermore, it was elucidated whether ANX AI overexpression affects migratory potential in Caki-1 cells. ANX AI immunohistochemistry was performed on 33 samples of CRCCs and 10 metastases. ANX AI expression was assessed in 12 samples by 2-dimensional gelelectrophoresis (2-DE), subsequent mass spectrometry and RT-PCR. Immunohistochemical data were statistically correlated with pathological parameters, amount of eosinophilic cells and clinical outcome. Furthermore, a haptotactic migration assay was done on Caki-1 cells transfected with ANX AI. Immunostaining for ANX AI was found in 18 tumours and all metastases investigated. Intensity of immunohistochemical staining correlated to Fuhrman grade, amount of eosinophilic cells and clinical outcome. 2-DE and RT-PCR confirmed the presence of ANX AI in neoplastic tissue. Overexpression of ANX AI did not significantly influence cell migration. From these findings ANX AI expression seems to be related to Fuhrman grade, clinical outcome and metastatic potential of CRCCs. Thus ANX AI could serve as a prognostic marker for tumour progression.

[In vivo imaging of the conjunctival epithelium using confocal laser scanning microscopy]. / In-vivo-Darstellung des Bindehautepithels. Eine vergleichende morphologische Untersuchung mittels konfokaler Laser-Scanning-Mikroskopie und Impressionszytologie.

BACKGROUND: In various ocular diseases, cytomorphological findings of the ocular surface are an essential component of clinical diagnostics. When evaluating the conjunctival epithelium, minimally invasive acquisition of biomaterial is necessary for lab and technical processing and in vitro histological examination. To examine corneal structures in vivo, confocal laser scanning microscopy is a successful standard method. Our aim was to employ in vivo confocal laser scanning microscopy also for examining the conjunctival epithelium. MATERIAL AND METHOD: Results were analyzed and compared with cytomorphological findings of impression cytology. Accordingly, the basic features of conjunctival in vivo examination using RLSM were described and defined. In vivo images were analyzed and compared with impression cytological slide preparations (n=110) of 23 healthy test persons. Examination was standardized. Finally, the confocal laser scan images were compared to the impression cytological patterns. RESULTS: Due to the distribution of reflectors (pixel brightness), diagnostic analysis of important morphological structures (cell nucleus, cytoplasm, nucleus/plasma relation) of the conjunctiva is possible. Secretory cells of the epithelium (goblet cells) can be easily recognized by their size. Highly reflective pixels depict cell walls or wide intercellular spaces with high contrast. CONCLUSIONS: The in vivo investigation of important anatomical and morphological structures of the conjunctival epithelium is possible using RLSM. The distribution pattern of goblet cell pixel brightness may correlate with various secretion contents or suggest distinct, recognizable, functional conditions (hypo- or hypersecretion).

Localization of annexins I, II, IV and VII in whole prostate sections from radical prostatectomy patients.

Annexins (ANXs) represent a family of calcium and phospholipid binding proteins that are involved in several physiological processes e.g. signal transduction, cellular differentiation and proliferation. Since they are known to be dysregulated in a variety of cancers we investigated the immunolocalization of ANXs in whole prostate sections containing benign prostatic epithelium (BPE), benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostate cancer (PCa) in order to evaluate their possible role during tumorigenesis. Samples were obtained from 28 patients undergoing radical prostatectomy. Gross sections of whole prostates were examined immunohistochemically for the distribution of ANX I, II, IV and VII. In BPE all ANXs were localized to the cell membranes and the cytoplasm of all gland cells. In BPH the immunoreactivity of ANX I and II was restricted to the basal cells of glands and expression pattern of ANX IV and VII was similar to BPE. In PIN only basal cells expressed ANX II. In PCa ANX II immunoreactivity was absent and weak ANX I and ANX IV immunoreactivity was restricted to the cytoplasm of tumor cells. ANX VII immunoreactivity was seen in some but not all tumor cells. Since ANX IV and VII expression did not show significant changes in PCa compared to non-neoplastic tissue and PIN an essential role during prostate tumourigenesis seems unlikely. In contrast, as progression from PIN to PCa is characterized by a reduction of ANX I and II this suggests that downregulation of these proteins could represent an important event in prostate carcinogenesis.

Expression of Skp2 and p27KIP1 in naevi and malignant melanoma of the skin and its relation to clinical outcome.

Skp2 (S-phase kinase associated protein 2) controls progression from G- to S-phase by promoting the proteolysis of the cyclin dependent kinase inhibitor p27KIP1. Despite the fact that a p27KIP1 decrease has been documented in melanoma progression, the role of Skp2 in these tumours is unknown. We therefore examined by immunohistochemistry the expression of Skp2, p27KIP1 and Ki-67 in 10 naevi (Ns), 15 superficial spreading melanomas (SSMs), 10 nodular melanomas (NMs) and 14 melanoma metastases (Ms). Nuclear Skp2 expression augmented with increasing malignancy (Ns: 1.4%, SSMs: 5.6%, NMs: 17.3%, Ms: 19.1%). In all tumours nuclear Skp2 expression correlated with Ki-67 (p=0.024) and inversely with p27KIP1 (p=0.007). A cytoplasmic reaction for Skp2 was also observed in most tumours and its expression decreased from Ns (12.3%) to SSMs (7.9%) and NMs (4.5%). In contrast, Ms showed an increase of cytoplasmic Skp2 (11.9%) that correlated with its nuclear expression (p=0.016). While nuclear Skp2 expression correlated with the pT-level (p=0.023), Clark-level (p=0.023) and Breslow index (p=0.019), the cytoplasmic Skp2 expression might be of biological significance only in NMs since it correlated with tumour depth (p=0.02) and pT-level (p=0.025). Our data suggests that Skp2 could contribute to melanoma progression. This is further highlighted by the fact that vertical growth phase (VGP) melanomas show significant higher nuclear Skp2 expressions when compared with the harmless radial growth phase (RGP) (p=0.047). Also nuclear Skp2 expression correlates with a reduced survival time (p=0.025) in melanoma.

[Molecular aspects of alterations in normal normal conjunctival epithelium. Role of apoptosis-associated genes]. / Molekulare Aspekte der Alterung des normalen Bindehautepithels Zur Bedeutung apoptoseassoziierter Gene.

BACKGROUND: The aim of the present study was to evaluate the age-dependent physiological expression pattern of apoptosis-related genes in the normal conjunctiva. PATIENTS AND METHODS: Healthy probands (group A: 24 probands and group B: 24 probands) were subjected to modified impression cytology using a PTFE membrane for evaluation of age-dependent expression of apoptosis-related genes in the normal conjunctiva. RNA was isolated from the cytologic impression specimens and used for cDNA synthesis employing SuperScriptTM II reverse transcriptase. RESULTS: RT-PCR revealed the detection of apoptosis-related mRNAs as follows (group A compared to group B): Apaf-1: 4.2/4.2%, Bcl-2: 41.7/41.7%, Bim: 0/0%, Bag-1: 70.8/100%, p53: 0/8.3%, Casp-3: 4.2/66.7%, Casp-5: 70.8/37.5%, Casp-8: 62.5/20.8%, Casp-9: 25/12.5%, c-myc: 83.3/95.8%, BAX: 91.7/70.8%, and BAD: 95.8/87.5%. CONCLUSION: The results revealed an age-dependent expression of apoptosis-associated genes (proapoptotic: Casp-3, Casp-5, Casp-8, antiapoptotic Bag-1) in normal human conjunctiva. Taken together these investigations provide basic information to understand the expression of apoptosis-associated genes in various diseases of the conjunctiva.

[Corneal anomalies in murine trisomy 16]. / Minoranomalien der Hornhaut bei der murinen Trisomie 16.

BACKGROUND: The prevalence of human Down's syndrome is about 1:700. Investigations using animal models are therefore of clinical relevance for understanding its etiopathogenesis. No corneal changes have been reported with transgenic murine trisomy 16. METHODS: A total of 20 fetal mice (n=40 eyes) with experimentally induced trisomy 16 were investigated from day 18 of pregnancy in order to determine whether visible developmental disorders of the cornea occur. All specimen were investigated microscopically in serial sections. RESULTS: In addition to disturbances in systemic development, the transgenic mouse fetuses showed high rates of malformation of the eyes. Developmental and differentiation disorders of the corneal epithelial cell layers and structural disturbances of the corneal parenchyma were found. Our findings are the first demonstration of developmental disorders of the cornea in mouse fetuses with trisomy 16. These minor anomalies of the cornea could well have resulted in keratoconus if the animals had survived. CONCLUSIONS: Our findings in transgenic mouse fetuses with trisomy 16 correspond to the clinical pattern of Down's syndrome in humans. Disturbed development of lids and lenses have a high prevalence, whereas corneal hypoplasia is found less often.

Expression of annexin II in conventional renal cell carcinoma is correlated with Fuhrman grade and clinical outcome.

Conventional renal cell carcinomas (CRCCs) were investigated for the expression of annexin II (ANX II) to determine out whether this calcium-binding protein could serve as a useful prognostic marker. CRCCs and adjacent nonneoplastic tissue from 33 patients were investigated for ANX II by immunohistochemistry, RT-PCR, and western blot analysis. ANX II expression was correlated with tumor differentiation (Fuhrman grade) and to clinical outcome. Tumors were composed of ANX II positive and negative cells. In grade I tumors only a weak membranous staining was seen in immunopositive cells. In grade II and III tumors, however, ANX II was seen in the cytoplasm and at the cell membranes of tumor cells. On serial sections membranous and cytoplasmic immunoreactivity for ANX II occurred predominantly in eosinophilic cells whereas clear cells were mostly immunonegative. The ANX II expression in CRCCs was correlated with clinical outcome and Fuhrman grade. Since ANX II expression is correlated with Fuhrman grade and clinical outcome it may be a useful marker for prognosis in CRCC.

Impact of genetic polymorphisms on the smoking-related risk of periodontal disease: the population-based study SHIP.

Periodontitis is a bacterial inflammatory disease leading to attachment loss with the consequence of tooth loss. There exists a multifactorial risk pattern including bacterial challenge, smoking, age, sex, diabetes, socio-economic and genetic factors. Smoking has the highest impact on the course of the disease modulated by all the other factors. Here, we report the relationship between smoking and the polymorphisms of genetic polymorphisms inflicted in the pathogenesis.In a randomly selected population-based study, 1083 subjects were typed for the polymorphisms of the IL-1 genotype, Fcgamma RIIIb receptor gene, myeloperoxidase and N-acetyltransferase (NAT2) and related to their periodontal state. Smoking behavior was assessed including present and past quality and quantity of smoking.There is a significant dose-effect relationship between the exposure to tobacco smoke and the extent of periodontal disease assessed as attachment loss and tooth loss. Moreover, there are gene-environmental interactions as subjects bearing variant genotypes show an enhanced smoking-associated risk of the disease modulated by these genotypes. In non-smokers, the impact of these genetic polymorphisms is mostly negligible.This study provides support for the hypothesis that subjects bearing genetic variants of polymorphically expressed phenotypes are at an increased risk of periodontitis when smoking. Mostly, this may be accomplished via the influence of smoking-related impairment on defense mechanisms rather than on the pathogenic pathways.

Expression of N-acetyltransferases in periodontal granulation tissue.

Smoking is a major risk of periodontal diseases. At the site of first contact, the gingiva is exposed to aromatic amines and polycyclic hydrocarbons. These are metabolized by the N-acetyltransferases (NAT), leading to local detoxification and/or activation reactions contributing to the risk of periodontal destruction in smokers. The purpose of this study was to detect the expression of N-acetyltransferase isoenzymes NAT1 and NAT2 in periodontal granulation tissue. In 24 specimens obtained from periodontitis patients or control subjects, mRNA encoding for NAT1 and NAT2 was detected by RT-PCR, and proteins were identified by immunohistochemistry. In periodontal granulation tissues, immunoreactivity for NAT1 and NAT2 was detected in infiltrating leukocytes and fibroblasts. In normal gingiva, both enzymes were found in epithelial cells, whereas NAT1 was also detected in endothelial cells. The results suggest that these enzymes may play a role in the defense against xenobiotics and the accelerated progression of periodontal disease in smokers.

A case of a persistent left vena cava superior with atresia of the right atrial ostium of the coronary sinus.

A persistent left vena cava superior with an atretic ostium of the coronary sinus was found during the routine dissecting course in the embalmed cadaver of an 83-year-old woman who had died from cardiac infarction. The left vena cava superior was very narrow in diameter (4 mm), originated at the lateral part of the left vena brachiocephalica and ran down between the venae pulmonales sinistrae and the auricula sinistra. The vena cava opened into the sinus coronarius of the heart, which terminated as a blind sac due to an atretic ostium. The vena coronaria sinistra as well as the vena interventricularis posterior drained into the sinus coronarius. Congenital atresia of the coronary opening is a rare malformation and is usually associated with other anomalies. The congenital ostial atresia could be the cause of a persistent left vena cava superior, which then takes over the drainage of the cardiac veins.

Glycodelin mRNA is expressed in the genital tract of male and female rats (Rattus norvegicus).

In the present study we demonstrate for the first time the expression of glycodelin mRNA in the female and male genital tracts of rats using non-radioactive in situ hybridisation. Glycodelin fragment 1 (+41 to +141) shares 100% homology with the human gene sequence. In the ovary, glycodelin mRNA was restricted to granulosa cells. In the uterus, glycodelin mRNA was expressed in all epithelial cells of the endometrium. In the male reproductive tract, glycodelin mRNA was distributed in all epithelial cells of the epididymis, the prostate and the seminal vesicle. However, in the testis, glycodelin mRNA was predominantly found in spermatogonia and in spermatocytes of the seminiferous epithelium. The expression in several reproductive organs of rats offers an excellent tool to study further the physiological role of glycodelin, which is so far thought to act as an immunosuppressive factor.