Localization Of Epidermal Growth Factor Research Articles

Effects of hCG on the electrical membrane properties of human trophoblast in culture

Cytologic localization of epidermal growth factor and its receptor in developing human placenta was analyzed by avidin-biotin immunoperoxidase techniques with a polyclonal antibody to epidermal growth factor and a monoclonal antibody to its receptor. In 4- to 5-week placenta, epidermal growth factor and its receptor were found to be almost exclusively localized to cytotrophoblasts, whereas in 6- to 12-week placentas they were predominantly localized to syncytiotrophoblasts. These findings suggest that both are initially expressed in cytotrophoblasts in very early placenta before 6 weeks' gestation and thereafter expressed in syncytiotrophoblasts in 6- to 12-week placentas. Their simultaneous expression in the cytotrophoblast of 4- to 5-week placentas and in the syncytiotrophoblast of 6- to 12-week placentas implies that epidermal growth factor may act in an autocrine manner in first-trimester placentas. By contrast, in second- and third-trimester placentas, epidermal growth factor was mainly localized to cytotrophoblasts, whereas its receptor was predominantly localized to syncytiotrophoblasts. These findings imply that epidermal growth factor may act in a paracrine fashion in second- and third-trimester placentas. The dynamic change in cytologic localization of epidermal growth factor and epidermal growth factor receptor in developing human placentas may reflect the change in a possible role of epidermal growth factor in the course of fetoplacental development.

Localization of epidermal growth factor and its receptor in mandibular molars of the rat prior to and during prefunctional tooth eruption.

Immunoperoxidase localization of epidermal growth factor receptors (EGFR) and epidermal growth factor (EGF) itself was examined in rat first and second mandibular molars postnatally from day 0 to 12. The results showed that the dental follicle stained heavily for EGFR from day 0 to 8, declined in staining at day 9, and was devoid of stain from day 10 onward. Preosteoblasts and osteoblasts of alveolar bone also stained and lesser staining of ameloblasts and odontoblasts was observed. Except for staining of occasional isolated cells, the stellate reticulum did not stain. Light staining of the dental pulp of the first mandibular molar was seen from day 0 onward but the pulp of the second molar did not stain until approximately day 6. With respect to EGF, the dental follicle also stained for it until day 12. The ameloblasts stained more intensely for EGF than for EGFR. Because injections of EGF cause premature eruption of teeth and because the presence of a dental follicle is necessary for eruption, this study suggests that EGF could have its effect on the follicle as seen by the presence of EGFR receptors on the follicle. Moreover, because EGF exerts its effects early (day 0-3) to cause eruption and because the influx of monocytes into the follicle to form osteoclasts for bone resorption for eruption occurs early, the heavy staining for EGFR in the follicle early followed by the absence of staining at day 10 correlates chronologically with the key molecular and cellular events of eruption. Finally, the presence of EGF in the follicle, as well as enamel organ, could provide an endogenous source of EGF to regulate tooth eruption, either by an autocrine or a paracrine effect. Thus, the localization of EGFR and EGF in the dental follicle coupled with the chronology of localization suggests that EGF could play a physiological role in tooth eruption.

Immunohistochemical localization of epidermal growth factor and its receptor during odontogenesis in the rat.

The expression of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFr) in developing teeth has been immunohistochemically studied in rat embryos (E-16 to E-21). Both EGF and EGFr showed a similar pattern of distribution. A very weak immunostaining was observed in the dental germ cells during the bud, cap, and bell teeth stages, as well as in few ectomesenchymal cells. In developed, but not erupted teeth, a moderate immunoreactivity for EGF and EGFr was present in the odontoblasts, in the ameloblasts and in the internal epithelial cells, but it was stronger in the dentine. In addition, the presence of EGF/EGFr was also observed in the intercalated ducts of salivary glands, primarily the submaxillary gland, in the maxillary bone cells, and in the cells of the peripheral and central nervous system. These results suggest that EGF has little or no effect during the early periods of tooth differentiation, whereas it is probably involved in the production of dentine. Moreover, EGF/EGFr seem to participate in the maturation and differentiation of other embryonic tissues such as tissues of the nervous system and bone.

Immunohistochemical Localization of Epidermal Growth Factor and Acidic and Basic Fibroblast Growth Factors in Postnatal Developing and Adult Rat Lungs

Histologic preparations of lungs form 1-, 5-, 10-, 18-, and 25-day-old rats and adult rats were probed immunohistochemically with specific antibodies for the distribution of epidermal growth factor (EGF), acidic fibroblast growth factor (aFGF), and basic fibroblast growth factor (bFGF). Immunoperoxidase staining of sections of adult rat lungs with a rabbit polyclonal antibody to bovine EGF was strong in ciliated cells of airways and mast cells, nonciliated cells of bronchioles, smooth muscle, type II cells of alveoli, and interstitial and epithelial cells of alveolar septal regions. Developing postnatal lungs had moderate and somewhat diffuse immunoreactivity in all epithelial cells, and more intense staining in vascular smooth muscle. Immunoperoxidase staining of adult rat lungs with a rabbit polyclonal antibody against bovine aFGF was distributed in identical fashion to EGF, with the exception of mast cells which were not reactive. These same sights were localized using an immunoperoxidase sequence with a rabbit polyclonal antibody to the leu 60-leu 98 fragment of aFGF (aFGFfr) with less background. In postnatal developing lungs, immunoreactivity with aFGF and aFGFfr preparations was diffuse and moderately intense in epithelial cells and vascular smooth muscle. Immunoperoxidase staining of adult rat lung sections with a monoclonal antibody to bovine bFGF was strong in hyaluronidase-digested preparations. Reactivity was principally confined to alveolar and vascular basement membrane regions and external laminae of smooth muscle. In early postnatal development, immunoreactivity for bFGF was found in basement membranes beneath developing epithelium and the endothelium of vessel wall intima and in the adventitia, with reactivity increasing with advancing age.(ABSTRACT TRUNCATED AT 250 WORDS)

A novel change in cytologic localization of human chorionic gonadotropin and human placental lactogen in first-trimester placenta in the course of gestation

Cytologic localization of human chorionic gonadotropin and human placental lactogen in developing human early placenta was analyzed by avidin-biotin immunoperoxidase techniques with an affinity-purified polyclonal antibody to beta-human chorionic gonadotropin carboxyl terminal peptide and a polyclonal antibody to human placental lactogen. In 4- to 5-week placentas human chorionic gonadotropin and human placental lactogen were found to be primarily localized to cytotrophoblasts, whereas in 6- to 12-week placentas these substances were exclusively localized to syncytiotrophoblast. We previously reported that a similar change in cytologic localization of epidermal growth factor and its receptor from cytotrophoblasts to syncytiotrophoblast in first-trimester placenta appeared between 5 and 6 weeks of gestation. Because epidermal growth factor was demonstrated to stimulate human chorionic gonadotropin and human placental lactogen production by early placental tissues, their simultaneous expression, as well as epidermal growth factor and its receptor in the cytotrophoblast of 4- to 5-week placenta and in the syncytiotrophoblast of 6- to 12-week placenta, implies that human chorionic gonadotropin and human placental lactogen production by first-trimester placenta may be regulated in an autocrine manner, wherein epidermal growth factor may serve as the signal. These findings suggest that in very early placenta, before 6 weeks of gestation, no sequential expression of human chorionic gonadotropin and human placental lactogen closely linked to syncytia formation may exist and that both can be expressed in the cytotrophoblast or undifferentiated stem cell of villous trophoblast in very early placenta.

Immunoelectron microscopic localization of epidermal growth factor in the eccrine and apocrine sweat glands.

We studied the localization of the epidermal growth factor (EGF) in eccrine and apocrine sweat glands with light microscopic and electron microscopic immunohistochemistry. Anti-human EGF (anti-hEGF) polyclonal antiserum and anti-hEGF monoclonal antibody (MAb) were used for the study. Light microscopic immunohistochemistry with monoclonal and polyclonal antibodies showed that hEGF-like immunoreactivity was strongly positive in the myoepithelial cells and weakly positive in the secretory cells of eccrine sweat glands. In apocrine sweat glands, it was strongly positive in the secretory cells as well as in the myoepithelial cells. Immunoelectron microscopy with polyclonal antibody showed that hEGF-like immunoreactivity was present in secretory granules of apocrine secretory cells. These granules had mitochondrion-like internal structure. No reactivity was observed on the eccrine secretory cells by immunoelectron microscopy. Neither dark cell granules nor mitochondria in eccrine secretory cells were labeled with anti-hEGF antibody. In both eccrine and apocrine sweat glands, hEGF-like immunoreactivity was diffusely present in the cytoplasm of myoepithelial cells. However, nuclei and mitochondria of myoepithelial cells were devoid of immunoreactivity for hEGF. Our observations indicate that apocrine sweat glands may secrete more hEGF in the sweat than eccrine sweat glands.

Localization of epidermal growth factor (EGF) binding sites on antiluminal plasma membrane of rat kidney: autoradiographic study using nonfiltering perfused rat kidney.

We previously demonstrated that the specific binding of EGF to the antiluminal plasma membrane was a prerequisite step for the renal uptake of EGF. In the present study, the localization of 125I-EGF binding sites on the antiluminal plasma membrane was investigated by tissue sampling and X-ray autoradiography in the nonfiltering kidney. The binding of 125I-EGF was recognized over the whole kidney and was highest in the inner medulla followed by the cortex and outer medulla. The binding of 125I-EGF in the nonfiltering kidney was completely inhibited in the presence of 20 nM unlabeled EGF, suggesting specific binding of 125I-EGF to its receptor. Further, we used a histologic tissue staining method to confirm the location of the 125I-EGF binding sites. Binding of 125I-EGF was demonstrated on the proximal straight tubules (PST), cortical collecting ducts (CCD), inner medullary collecting ducts (IMCD), and thin limb of Henle in the inner medulla (IMTLH). We found that the binding of 125I-EGF was high in the IMTLH. In addition, we determined the grain density both on the cell surface membrane and in the intracellular space of the proximal straight tubules, where the grain density on the antiluminal plasma membrane was approximately 50% that in the intracellular space at 20 min after the start of 125I-EGF perfusion, suggesting the internalization of 125I-EGF from the antiluminal plasma membrane to the intracellular compartment. In conclusion, the binding sites of 125I-EGF, which were accessible from the antiluminal side, were broadly distributed over the whole kidney and were most dense around the IMTLH.

Cytologic localization of epidermal growth factor and its receptor in developing human placenta varies over the course of pregnancy

Cytologic localization of epidermal growth factor and its receptor in developing human placenta was analyzed by avidin-biotin immunoperoxidase techniques with a polyclonal antibody to epidermal growth factor and a monoclonal antibody to its receptor. In 4- to 5-week placenta, epidermal growth factor and its receptor were found to be almost exclusively localized to cytotrophoblasts, whereas in 6- to 12-week placentas they were predominantly localized to syncytiotrophoblasts. These findings suggest that both are initially expressed in cytotrophoblasts in very early placenta before 6 weeks' gestation and thereafter expressed in syncytiotrophoblasts in 6- to 12-week placentas. Their simultaneous expression in the cytotrophoblast of 4- to 5-week placentas and in the syncytiotrophoblast of 6- to 12-week placentas implies that epidermal growth factor may act in an autocrine manner in first-trimester placentas. By contrast, in second- and third-trimester placentas, epidermal growth factor was mainly localized to cytotrophoblasts, whereas its receptor was predominantly localized to syncytiotrophoblasts. These findings imply that epidermal growth factor may act in a paracrine fashion in second- and third-trimester placentas. The dynamic change in cytologic localization of epidermal growth factor and epidermal growth factor receptor in developing human placentas may reflect the change in a possible role of epidermal growth factor in the course of fetoplacental development.

Cytologic localization of epidermal growth factor and its receptor in developing human placenta varies over the course of pregnancy

Cytologic localization of epidermal growth factor and its receptor in developing human placenta was analyzed by avidin-biotin immunoperoxidase techniques with a polyclonal antibody to epidermal growth factor and a monoclonal antibody to its receptor. In 4- to 5-week placenta, epidermal growth factor and its receptor were found to be almost exclusively localized to cytotrophoblasts, whereas in 6- to 12-week placentas they were predominantly localized to syncytiotrophoblasts. These findings suggest that both are initially expressed in cytotrophoblasts in very early placenta before 6 weeks' gestation and thereafter expressed in syncytiotrophoblasts in 6- to 12-week placentas. Their simultaneous expression in the cytotrophoblast of 4- to 5-week placentas and in the syncytiotrophoblast of 6- to 12-week placentas implies that epidermal growth factor may act in an autocrine manner in first-trimester placentas. By contrast, in second- and third-trimester placentas, epidermal growth factor was mainly localized to cytotrophoblasts, whereas its receptor was predominantly localized to syncytiotrophoblasts. These findings imply that epidermal growth factor may act in a paracrine fashion in second- and third-trimester placentas. The dynamic change in cytologic localization of epidermal growth factor and epidermal growth factor receptor in developing human placentas may reflect the change in a possible role of epidermal growth factor in the course of fetoplacental development. Cytologic localization of epidermal growth factor and its receptor in developing human placenta was analyzed by avidin-biotin immunoperoxidase techniques with a polyclonal antibody to epidermal growth factor and a monoclonal antibody to its receptor. In 4- to 5-week placenta, epidermal growth factor and its receptor were found to be almost exclusively localized to cytotrophoblasts, whereas in 6- to 12-week placentas they were predominantly localized to syncytiotrophoblasts. These findings suggest that both are initially expressed in cytotrophoblasts in very early placenta before 6 weeks' gestation and thereafter expressed in syncytiotrophoblasts in 6- to 12-week placentas. Their simultaneous expression in the cytotrophoblast of 4- to 5-week placentas and in the syncytiotrophoblast of 6- to 12-week placentas implies that epidermal growth factor may act in an autocrine manner in first-trimester placentas. By contrast, in second- and third-trimester placentas, epidermal growth factor was mainly localized to cytotrophoblasts, whereas its receptor was predominantly localized to syncytiotrophoblasts. These findings imply that epidermal growth factor may act in a paracrine fashion in second- and third-trimester placentas. The dynamic change in cytologic localization of epidermal growth factor and epidermal growth factor receptor in developing human placentas may reflect the change in a possible role of epidermal growth factor in the course of fetoplacental development.

Immunohistochemical localization of epidermal growth factor (EGF) and EGF receptor in human oral mucosa and its malignancy

The immunohistochemical localizations of human epidermal growth factor (hEGF) and EGF receptor (EGFr) in oral tissues, including normal mucosa, leukoplakia and squamous cell carcinoma were examined by the use of monoclonal antibodies to hEGF and EGFr. In normal mucosa and leukoplakia, immunostaining of hEGF was limited to an underlying layer of connective tissue near the epithelium. The intensity of extracellular staining appeared to increase with the degree of epithelial malignancy and was eventually most striking in the stroma of invasive carcinoma. The epithelial cells in normal mucosa, leukoplakia, and squamous cell carcinoma showed negligible immunoreactivity for hEGF. Expression of EGFr appeared to be associated with the proliferative activity of cells and/or epithelial malignancy. In normal mucosa, anti-EGFr monoclonal antibody reacted only with the basal cell layer. In all sections of leukoplakia, the positive cells for EGFr were found in the prickle cell layer in addition to the basal cell layer. Most tumour cells in squamous cell carcinoma were strongly positive for EGFr. These findings indicate increased expression of hEGF and EGFr with malignancy. The characteristic localization of extracellular hEGF in the underlying connective tissue and in stroma of oral mucosal tumours suggests a possible epithelial-mesenchymal interaction in hEGF secretion.

Bidirectional transport of selenium in the perfused human placenta in vitro

Cytologic localization of human chorionic gonadotropin and human placental lactogen in developing human early placenta was analyzed by avidin-biotin immunoperoxidase techniques with an affinity-purified polyclonal antibody to β-human chorionic gonadotropin carboxyl terminal peptide and a polyclonal antibody to human placental lactogen. In 4- to 5-week placentas human chorionic gonadotropin and human placental lactogen were found to be primarily localized to cytotrophoblasts, whereas in 6- to 12-week placentas these substances were exclusively localized to syncytiotrophoblast. We previously reported that a similar change in cytologic localization of epidermal growth factor and its receptor from cytotrophoblasts to syncytiotrophoblast in first-trimester placenta appeared between 5 and 6 weeks of gestation. Because epidermal growth factor was demonstrated to stimulate human chorionic gonadotropin and human placental lactogen production by early placental tissues, their simultaneous expression, as well as epidermal growth factor and its receptor in the cytotrophoblast of 4- to 5-week placenta and in the syncytiotrophoblast of 6- to 12-week placenta, implies that human chorionic gonadotropin and human placental lactogen production by first-trimester placenta may be regulated in an autocrine manner, wherein epidermal growth factor may serve as the signal. These findings suggest that in very early placenta, before 6 weeks of gestation, no sequential expression of human chorionic gonadotropin and human placental lactogen closely linked to syncytia formation may exist and that both can be expressed in the cytotrophoblast or undifferentiated stem cell of villous trophoblast in very early placenta.

Detection of Epidermal Growth Factor Receptors and E‐Cadherins in the Basolateral Membrane of A431 Cells by Laser Scanning Fluorescence Microscopy

We have examined the localization of epidermal growth factor (EGF) receptors and E‐cadherins in cultured A431 cells, an epidermoid carcinoma cell line, using a laser scanning fluorescence microscope. The fluorescence signals generated by monoclonal antihodies against EGF receptor and E‐cadherin were localized mainly in the cell‐cell contact sites, about 3 μm in width. When these areas for cell adhesion were scanned perpendicularly, fluorescence was detected from a point 4μ from the cell base for a distance of 8 μm towards the cell surface (about 16 μm from the cell base). These data suggest the subcellular localization of EGF receptors and E‐cadherins in the basolateral membrane of A431 cells.

The correlation of epidermal growth factor with invasion and metastasis in human gastric cancer

We examined the localization of epidermal growth factor (EGF) in 185 specimens of primary human gastric cancer using the avidin-biotin peroxidase complex immunohistochemical method on formalin-fixed paraffin-embedded sections. Thirty-four per cent of the gastric cancer specimens were positive for EGF, which was mainly located in the cytoplasm of the cancer cells and occasionally in the stromal cells, but was not detected in non-cancerous gastric epithelium. Moreover, the presence of EGF in gastric cancer was correlated with gastric wall invasion and lymph node metastasis. EGF was found more often in advanced cancers than in early ones (p less than 0.01), and also more often in cancers with lymph node metastasis than in those without (p less than 0.05). The five-year survival of patients with EGF-positive tumors was worse than that of patients with EGF-negative tumors (p less than 0.05). The presence of EGF in human gastric cancer may thus represent higher malignant potential.

Immunohistochemical demonstration of epidermal growth factor in human gastric cancer xenografts of nude mice

Thirty-two surgical specimens and three cell lines of human gastric cancers were used for subcutaneous transplantation into nude mice, resulting in the establishment of eight (25%) xenografts from the surgical specimens and two (67%) from the cell lines. The localization of epidermal growth factor (EGF) in the surgical specimens and cell lines of the gastric cancers and their xenografts in nude mice was then investigated immunohistochemically. Epidermal growth factor was stained in the cytoplasm of the cancer cells, being detected in 16 (50%) of the 32 surgical specimens and in all of the cell lines. Seven (44%) of the sixteen EGF-positive surgical specimens and one (6%) of the 16 EGF-negative ones were tumorigenic in nude mice. All of the xenografts in nude mice were positive for EGF. The tumorigenicity of human gastric cancer xenografts in nude mice may, therefore, be correlated with the presence of EGF in cancer cells.

EPIDERMAL GROWTH FACTOR IN CHOLESTEATOMA

The localization of epidermal growth factor (EGF) in human cholesteatoma tissue was examined immunohistochemically, using sections of formalin-fixed, paraffin-embedded tissue with avidin-biotin peroxidase complex method. Thirty-three cases of active cholesteatoma which had abundant granulation and debris were studied. Out of 33 cases, EGF was positive in 32 cases (97%) in the epidermis, 30 cases (91%) in the fibroblast, and 21 cases (64%) in the endothelial cells. In 9 cases of inactive cholesteatoma, on the other hand, EGF was positive in 5 cases (56%) in the epidermis, no case in the fibroblast, and a case (11%) in the endothelial cells. Active cholesteatoma had higher immunoreactivity in the epidermis than inactive cholesteatoma. This suggests that the activity of cholesteatoma is indicated by the immunoreactivity of EGF. Difference of EGF immunoreactivity between active and inactive cholesteatoma was bigger in the fibroblast in the subcutaneous tissue of cholesteatoma than in the epidermis. This gives the reason that the activity of cholesteatoma exists in the subcutaneous tissue. These results suggest that EGF plays an important role in accelerating the growth of cholesteatoma.

EPIDERMAL GROWTH FACTOR IN CHOLESTEATOMA

The localization of epidermal growth factor (EGF) in human cholesteatoma tissue, normal ear drum and external auditory canal skin was examined immunohistochemically, using avidin-biotin peroxidase complex method. Bouin-fixed tissue was stained for investigation of horny layer in the epidermis, because fixation in Bouin's solution provides better preservation of the antigen. In the horny layer of cholesteatoma tissue, 19 out of 24 cases had EGF-positive immunoreactivity (79%). In 2 cases of normal external auditory canal skin, 4 cases of normal ear drum and a case of postauricular skin, no EGF-immunoreactivity was revealed in the horny layer. EGF was assayed in the debris of cholesteatoma and the horny layer of the normal bony external canal with dot blot immunoassay. EGF content of the debris was higher than that of the horny layer of normal skin. The result of the first report suggests the activity of cholesteatoma exists in the subcutaneous tissue (see the previous paper). In this report EGF content of cholesteatoma in the horny layer was found higher than that of normal external skin. This result demonstrates that EGF in the horny layer plays an important role in accelerating the growth and bony destruction in cholesteatoma. To summarize these two reports, the following conclusion was reached. In the epidermis EGF content is equal in cholesteatoma and normal skin. But in the subcutaneous tissue and the horny layer EGF content of cholesteatoma is higher than that of normal skin. EGF in situ may be strongly related to the growth and bony destruction of cholesteatoma.

Nonrandom distribution of epidermal growth factor receptors on the plasma membrane of human A431 cells

The localization of epidermal growth factor (EGF) receptors over the plasma membranes of human epidermoid carcinoma A431 cells was analyzed at the electron microscopic level using surface replica techniques and conventional thin sections, in combination with immunocytochemistry. Immunolabeling was performed using two distinct monoclonal antibodies directed against the extracellular portion of the receptor, followed by protein A-colloidal gold conjugates. Unexpectedly, with the first monoclonal antibody used, the distribution of the receptors in both unfixed and glutaraldehyde-fixed cells was clearly regionalized, showing a preferential localization of the immunolabeling at the cell periphery as well as over the areas rich in microvilli and in coated and uncoated pits. A similar pattern of distribution was observed also with the other monoclonal antibody, but only when the cells were fixed with glutaraldehyde before immunolabeling. Treatment with the phorbol ester 12- O-tetradecanoylphorbol-13-acetate modifies this distribution, inducing a more disperse pattern. Our observations suggest that a minor group of EGF receptors, which may represent the high-affinity receptors, presents a regional distribution, similar to that described for typical recycling receptors.

Progestin and estrogen receptors: characterization and localization in rat submandibular glands, with special reference to epidermal growth factor.

By using progestin (P) and estrogen (E), the localization and characterization of both steroid receptors were examined in the submandibular gland (SMG) of 6-week-old immature castrated rats, with special reference to localization of epidermal growth factor (EGF). In the castrated male and female rats, both 3H-estradiol-17 beta (3H-E2 beta) and 3H-promegestone (3H-R5020) bound to SMG cytosol with high affinity and low capacity. These values were similar to those reported for other tissues. However, E-treatment after castration inhibited the specific binding. In sucrose density gradient ultracentrifugation, it was found that P receptors in both castrated males and females had a sedimentation coefficient of 7S, whereas E receptors had sedimentation coefficients of 4S and 7S. A histochemical study of the SMG of castrated male and female rats showed that the E-peroxidase complex (EPC)- and P-peroxidase complex (PPC)-stained cells were predominantly located in the epithelium of the duct system including the excretory duct and the granular convoluted tubules. Few cells were located in the intercalated duct, and none were found in the acinus. EGF-immunoreactive cells were also located in the epithelium of the same tissue region as in PPC- and EPC-stained sections. Moreover, E-treatment after castration inhibited the intensity of staining and immunoreactivity. These results clearly suggest that rat SMG contains specific P and E receptors which are mainly located in the epithelial cells of the duct system in which EGF-containing cells are identified. We discussed the possibility that P and E might affect EGF immunoreactivity, which reflects EGF production, through their receptors in the epithelium of the duct system.

Human corpus luteum: immunocytochemical localization of epidermal growth factor

Six corpora lutea (day 17 to day 25) of the menstrual cycle and four ovarian stromal tissues from seven cycling women were examined for the presence of epidermal growth factor (EGF) by immunocytochemistry with the use of the indirect peroxidase-antiperoxidase (PAP) procedure. After tissue sections of 4 micron were mounted, endogenous peroxidases were removed with hydrogen peroxide, and the sections were incubated for 1 hour at room temperature followed by 16 hours at 4 degrees C with a highly specific antisera for mouse EGF (mEGF), nonimmunized normal rabbit serum, or antiserum preadsorbed with excess antigen. After the reaction with the second antibody (goat antirabbit IgG) for 1 hour at room temperature, the peptide was localized by use of PAP and 3.3'-diaminobenzidine as the chromogen. EGF could be localized in the luteal cells of five corpora lutea (day 17 to day 25) but not in a corpus luteum of day 22 and all ovarian stromal tissue examined. The localization of immunoreactive EGF in the human corpus luteum demonstrates directly for the first time the presence of this growth-promoting peptide in this tissue, which suggests its local production.

Immunocytochemical localization of epidermal growth factor in mouse kidney.

Epidermal growth factor (EGF) was originally isolated from mouse submandibular glands (SMG). However, SMG removal failed to lower circulating EGF, and large amounts of EGF have been found in mouse urine. In addition, the presence of pre-pro-EGF mRNA in mouse kidney has recently been reported by others. Kidneys may therefore represent an alternate source of EGF. In the present study, we investigated the immunocytochemical localization of EGF in mouse kidney. Male and female adult Swiss Webster mice were fixed by perfusion with 4% paraformaldehyde or Zamboni's fixative, the kidneys were frozen, and serial sections were obtained. Rabbit EGF antiserum was used for the primary incubation and the avidin-biotin complex immunoperoxidase procedure was utilized for immunostaining. EGF was immunolocalized in the apical portion of the cells lining the thick ascending limb of Henle (TALH) and the distal convoluted tubule (DCT). The macula densa, in contrast, lacked EGF immunoreactivity. No sex differences were observed in the distribution pattern or intensity of immunostaining. Infusion of EGF into sheep renal artery has been reported to induce changes in urine flow and ionic composition. Immunolocalization of EGF in the TALH and DCT documented here supports a regulatory role for EGF in the function of the mouse distal nephron.