Arteriolar Endothelial Cells Research Articles

Expression of matrilysin in vascular endothelial cells adjacent to matrilysin-producing tumors.

Matrilysin is believed to play important roles in tumor progression and metastasis. In the present study, we analyzed matrilysin-producing cells in various human cancer tissues by immunohistochemistry and in situ hybridization. Tumor cells in colorectal carcinomas, pancreatic carcinomas, transitional-cell carcinomas of the kidney and small-cell lung carcinomas were frequently positive for matrilysin. In addition, we found that endothelial cells of arterioles and venules adjacent to matrilysin-positive tumors expressed matrilysin mRNA and protein. The endothelial cells adjacent to matrilysin-negative tumors and those in normal tissues were negative for matrilysin. Furthermore, analyses by casein zymography, Western blotting and RT-PCR showed that matrilysin was weakly expressed by cultured human umbilical vein endothelial cells. Our results suggest that the expression of matrilysin in vascular endothelial cells and in tumor cells may be regulated by common soluble factors, and that endothelial cell-derived matrilysin may contribute to tumor angiogenesis and tumor metastasis.

Cellular distribution of 5‐lipoxygenase and leukotriene receptors in postsurgical peritoneal wound repair

Products of the eicosanoid pathways, namely prostaglandins and leukotrienes, are known to play a key role in inflammatory and immune responses, as well as other wound cellular activities of wounded tissue. The objective of this study was to determine whether surgically induced intraperitoneal and incisional wounds in the rat express 5-lipoxygenase and contain binding sites for leukotrienes and whether their levels change during the course of healing. With the use of a specific monoclonal antibody generated against 5-lipoxygenase, the enzyme was immunohistochemically localized in various wound cells during postsurgical days 2 to 35. The inflammatory cells within the wound were the major cell types containing 5-lipoxygenase immunoreactive protein, followed by fibroblasts in the incisional and peritoneal fibrous adhesions, striated muscle, and the vasculature. The greatest level of immunostaining was observed during the first 2 weeks after surgery, which decreased to near unwounded levels by day 35. Light microscope autoradiographic binding studies using (3)H-leukotrienes indicated that the peritoneal/incisional wounds and unwounded tissues contain specific (3)H-leukotriene C(4) and (3)H-leukotriene D(4) but not (3)H-leukotriene B(4) binding sites. Quantitative grain analysis (net grain density/100 microm(2)), representing specific (3)H-leukotriene C(4) and (3)H-leukotriene D(4) binding sites calculated for different cell types in the wound and unwounded regions showed that (3)H-leukotriene C(4) binding was highest over the striated muscle proximal to the injury and incisional and peritoneal granulation tissue fibroblasts. The net grain density over these cells increased by 3-, 2.5-, and 2-fold by day 14, respectively, and declined to the control values by day 21 after injury (p < 0.05). The pattern of (3)H-leukotriene D(4) binding was similar to that observed for (3)H-leukotriene C(4), but with a lower density. The grain density for (3)H-leukotriene C(4) and (3)H-leukotriene D(4) in arteriolar endothelial and smooth muscle cells remained unchanged. These data suggest that the products of the lipoxygenase pathway through the presence of their specific receptors may play an important role in peritoneal wound repair and adhesion formation.

Elevation of intracellular calcium in smooth muscle causes endothelial cell generation of NO in arterioles.

It is well known that vascular smooth muscle tone can be modulated by signals arising in the endothelium (e.g., endothelium-derived relaxing factor, endothelium-derived hyperpolarizing factor, and prostaglandins). Here we show that during vasoconstriction a signal can originate in smooth muscle cells and act on the endothelium to cause synthesis of endothelium-derived relaxing factor. We studied responses to two vasoconstrictors (phenylephrine and KCl) that act by initiating a rise in smooth muscle cell intracellular Ca2+ concentration ([Ca2+]i) and exert little or no direct effect on the endothelium. Fluo-3 was used as a Ca2+ indicator in either smooth muscle or endothelial cells of arterioles from the hamster cheek pouch. Phenylephrine and KCl caused the expected rise in smooth muscle cell [Ca2+]i that was accompanied by an elevation in endothelial cell [Ca2+]i. The rise in endothelial cell [Ca2+]i was followed by increased synthesis of NO, as evidenced by an enhancement of the vasoconstriction induced by both agents after blockade of NO synthesis. The molecule involved in signal transmission from smooth muscle to endothelium is as yet unknown. However, given that myoendothelial cell junctions are frequent in these vessels, we hypothesize that the rise in smooth muscle cell Ca2+ generates a diffusion gradient that drives Ca2+ through myoendothelial cell junctions and into the endothelial cells, thereby initiating the synthesis of NO.

High levels of human chymase expression in the pineal and pituitary glands

The brain renin-angiotensin system plays a role in both cardiovascular homeostasis and neurosecretory functions. Since the mechanisms of angiotensin (Ang) II formation in the human brain have not been clarified, the aims of the present study were to determine the presence of human chymase and angiotensin I-converting enzyme (ACE) in human and non-human brains. In the human brain, the total Ang II-forming activity was significantly higher in the pineal and pituitary glands than those in other regions. In other species (rat, bovine and porcine), the level of chymase as well as total Ang II-forming activities in pineal glands were significantly lower than those in human glands. High levels of chymase-like immunoreactivity (ir) were found in the arteriolar endothelial cells, adventitial mesenchymal cells and in parenchymal cells of the human pineal and pituitary glands while ACE-ir was mostly observed in the endothelial cells and occasionally found in parenchymal cells. Our study provides the first evidence that human chymase exists in the pineal and pituitary glands. The remarkable regional and species differences in mechanisms of Ang II formation suggest a specific role of chymase or ACE in the human brain.

Primary Pulmonary Hypertension and Human Immunodeficiency Virus Infection

This report details the case of a 42-year-old homosexual Caucasian male with infection due to human immunodeficiency virus type 1 (HIV-1) who presented with a four-month history of progressive dyspnea and was found to have clinical and hemodynamic evidence of severe pulmonary hypertension. He had had no opportunistic infections, and had a T helper lymphocyte count of 200×10(6)/L. Extensive clinical laboratory and radiological evaluations revealed no underlying cause. Microscopic examination of postmortem lung tissue revealed findings consistent with grade V pulmonary hypertension. Electron microscopic analysis and polyermase chain reaction detection of HIV-DNA from dissected pulmonary arterioles failed to provide any supportive evidence to suggest productive infection of the pulmonary arteriolar endothelial cells by HIV-1. Although HIV-1 likely plays a role in the pathogenesis of primary pulmonary hypertension, evidence for direct infection of pulmonary vessel endothelium was lacking in this case. The pathogenesis of primary pulmonary hypertension associated with HIV remains obscure.

Chronological immunohistochemical detection and localization of porcine reproductive and respiratory syndrome virus in gnotobiotic pigs.

An immunogold-silver immunohistochemical technique was used to determine the chronological distribution and localization of porcine reproductive and respiratory syndrome virus (PRRSV) in experimentally infected gnotobiotic pigs. Thirty-two pigs were randomly allocated to infected (n = 24) or control (n = 8) groups. Pigs in infected groups were inoculated at 3 days of age by nasal instillation of PRRSV isolate ATCC VR-2332 (total dose = 10(2.64) TCID50), and control pigs were exposed in the same manner to uninfected cell culture supernatant. Three infected and one control pigs were euthanatized at 12 hours and at 1, 2, 3, 5, 7, 14, and 21 days postexposure (DPE). Bronchiolar epithelial cells, arteriolar endothelial cells, monocytes, and interstitial, alveolar, and intravascular macrophages stained for PRRSV antigen at 12 hours postexposure. Staining for PRRSV antigen in endothelial cells, monocytes, and alveolar, interstitial, and intravascular macrophages was most intense and widespread in lung sections from 14 and 21 DPE. In the heart, macrophages in the interstitial and subendocardial spaces and endothelial cells in a few arterioles stained for PRRSV antigen at 14 and 21 DPE. Tonsillar macrophages and mucosal epithelium stained for PRRSV antigen at 12 hours postexposure and sporadically with less intensity in subsequent sampling periods. In the nasal turbinate, PRRSV antigen was identified in macrophages within the mucosal epithelium at 12 hours postexposure and again at 14 and 21 DPE. There was focal staining for PRRSV antigen in the choroid plexus in one pig at 14 DPE. Based on the results of this experiment, the pathogenesis of PRRSV infection in gnotobiotic pigs can be described as initial virus entry through nasal epithelial, tonsillar, and pulmonary macrophages, with viremia occurring by 12 hours postexposure followed by the development of pneumonia, myocarditis, encephalitis, rhinitis, vasculitis, and lymphoid necrosis. Although PRRSV can infect macrophages in heart, tonsil, turbinate, and choroid plexus, pulmonary macrophages are predominantly and consistently infected and are the predominantly cells for virus replication in gnotobiotic pigs.

Hypoxia activates nitric oxide synthase and stimulates nitric oxide production in porcine coronary resistance arteriolar endothelial cells

Hypoxia activates nitric oxide synthase and stimulates nitric oxide production in porcine coronary resistance arteriolar endothelial cells

Hypoxia activates nitric oxide synthase and stimulates nitric oxide production in porcine coronary resistance arteriolar endothelial cells

Hypoxia significantly alters vascular tone in coronary resistance arterioles during prolonged ischemia, potentially through the modulation of endothelial cell metabolism as well as endothelial function. The objective of this study was to test the hypothesis that constitutive nitric oxide synthase (cNOS) is sensitive to oxygen tension and that hypoxia increases the activity of cNOS and nitric oxide production in the porcine coronary microcirculation. Monocultures of porcine coronary resistance arteriolar endothelial cells (RAEC) were isolated and proven to be endothelium based upon morphology, binding of acetylated LDL, and factor VIII antigen positivity. Cells were exposed to either hypoxia (pO2 = 10 mmHg) or normoxia (pO2 = 160 mmHg) for varying periods of time. Nitric oxide production was directly measured using a chemiluminescence method, while cNOS enzyme activity was assayed using a fibroblast-report cell method. cNOS protein was quantitated by Western blot analysis using the H32 monoclonal antibody to the endothelial cell constitutive isoform of NOS. Hypoxia significantly augmented A23187-stimulated nitric oxide production [23.77 (1.73) vs 14.94 (0.66) nmol . micrograms-1 protein, hypoxia vs. normoxia respectively, n = 8, P < 0.01]. Using the fibroblast reporter cell assay, cNOS activity was increased in RAEC after exposure to hypoxia for 30, 120 and 240 min [normoxia control: 0.16 (0.04) fmol . microgram-1 protein; hypoxia: 30 min = 1.00 (0.19), 120 min = 1.08 (0.04), 240 min = 1.26 (0.07) fmol . micrograms-1 protein (n = 6, p < 0.01)]. Western blots showed a single band at 135 kDa that was increased in homogenates of cells previously exposed to hypoxia. These experiments demonstrated that the regulation of cNOS is sensitive to oxygen tension. Hypoxia significantly activated constitutive nitric oxide synthase in coronary resistance arteriolar endothelial cells, and this was translated to an increased production of nitric oxide.

Carbohydrate cytochemistry of the endothelium lining the splenic blood vessels in the rat.

In the endothelial cells lining the rat splenic blood vessels, neutral carbohydrates were studied by means of combined periodic acid-thiocarbohydrazide-silver protein (PA-TCH-SP) and alpha-amylase digestion methods. In the endothelial cells lining the central and follicular arteries of the spleen, the neutral glycoconjugate-containing surface coat of the luminal plasma membrane and related pinocytotic invaginations and vesicles in the apical cytoplasm were strikingly distinguished, as compared with those in the cells lining the splenic sinuses. In contrast, cytoplasmic glycogen particles in the sinus endothelial cells were apparently larger in amount than those in the arteriolar endothelial cells. Such cytochemical variations of neutral carbohydrates with the arteriolar and venous vessels of the rat spleen were discussed with special reference to varying cytophysiological functions of the endothelial cells with the different segments of the splenic blood vessels.

Expression of nitric oxide synthase immunoreactivity by interstitial cells of the canine proximal colon

A subpopulation of interstitial cells (ICs) are interposed between nerve terminals and smooth muscle cells in the gastrointestinal tract and may participate in neuromuscular transmission. These cells appear to be targets for NO released from enteric inhibitory nerves and respond to exogenous NO with: (i) an elevation in cGMP levels; (ii) an increase in intracellular Ca 2+; (iii) and release of a diffusible substance that has tentatively been identified as NO. For the latter to be possible, ICs must express a constitutive isoform of NOS. This study characterized the expression of NOS-like immunoreactivity (NOS-LI) in ICs of the canine colon using 3 antibodies raised against the 2 known constitutive forms of NOS (i.e., neural (nNOS) and endothelial (eNOS) isoforms). Antibodies raised against eNOS and an antibody raised against rat cerebellar nNOS labeled ICs along the submucosal surface of the circular muscle layer (IC-SM), along the surface of septa that separate the circular muscle into fiber bundles (IC-SM), and in the myenteric region between the circular and longitudinal muscle layers (IC-MY). Another antibody raised against rat cerebellar nNOS failed to label ICs. Cultured IC-SM also expressed NOS-LI, suggesting that this feature of the IC phenotype survives culture conditions. Arteriolar endothelial cells in the canine colon were labeled with the same 2 antibodies that labeled ICs, suggesting there are significant structural similarities between NO synthases in ICs and endothelial cells. The data suggest that IC-SM and IC-MY express a constitutive form of NOS. Synthesis of NO by ICs may influence electrical rhythmicity and may serve to amplify and even propagate enteric inhibitory neurotransmission.

A novel application of microsphere perfusion and scanning electron microscopy to the identification of pulmonary arterioles in guinea‐pig and rabbit lungs

In arterioles of the lung the intravascular blood pressures are lower than in comparable vessels in the systemic circulation and the arteriole walls are thinner. Therefore, it is very difficult to distinguish between arterioles and venules of the same size using scanning electron microscopy. This study describes a novel application of latex microsphere perfusion and scanning electron microscopy which distinguishes between pulmonary arterioles and venules on the basis of endothelial cell morphology. Microspheres, 90 and 45 microns in diameter, were perfused into the arterial side of the pulmonary circulation of guinea-pig and rabbit lungs. Scanning electron microscopy of the arterioles on both sides of the lodged microspheres indicated that the endothelial cells are spindle shaped. In contrast, the endothelial cells of equal diameter venules are polygonal. Furthermore, the nuclei of the arteriolar endothelial cells were significantly (P = 0.019) narrower than those of endothelial cells in venules of equal diameter. Finally, it was observed that the differences between arteriole and venule endothelial cells persisted distally to the capillaries.

Immunohistochemical Localization of Insulin-like Growth Factor (IGF-I), IGF-I Receptor, and IGF Binding Proteins 1–4 in Human Fallopian Tube at Various Reproductive Stages

Despite the existence of numerous data regarding the insulin-like growth factor (IGF) system in reproductive tissues, especially uterine and ovarian, very little information is available concerning their presence in oviductal/fallopian tube tissue. To elucidate this, the present study was undertaken to determine the presence and cellular distribution of IGF-I, IGF-I receptor (IGF-IR), and IGF binding proteins (IGFBP) 1-4 in human fallopian tubes during various reproductive stages, by means of immunohistochemical analysis with specific antibodies to IGF-I, IGF-IR and IGFBPs 1-4. The primary site of immunoreactivity for these proteins in fallopian tube tissue was in the epithelial lining of the tubes, with substantially lower intensity in the smooth muscle layer, fibroblasts of the serosal tissue, and arteriolar endothelial and smooth muscle cells. The immunostaining was associated with both ciliated and nonciliated tubal epithelial cells without substantial differences in their intensity. There were also no differences in immunoreactivity of IGF-I, IGF-IR, and IGFBPs 1-4 present in ampullary vs. isthmus region of the tubes. Specimens obtained 5-12 years post tubal ligation stained similarly to sections from unligated tubes taken during the same phase of the cycle. The intensity of immunostaining for the IGFBPs was greatest for IGFBP-1, followed by IGFBP-4, then IGFBP-2 and IGFBP-3, with 2 and 3 having similar intensities. The immunostaining intensity of IGF-I, IGF-IR, and IGFBPs in fallopian epithelial cells was cycle-dependent and considerably higher in late proliferative and early-mid secretory compared to late secretory phases, with little immunostaining in the early proliferative phase of the menstrual cycle and postmenopausal period. In summary, the immunohistochemical study reported here demonstrates the presence of IGF-I, IGF-IR, and IGFBPs 1-4 in the human fallopian tube during various reproductive stages. These data suggest an autocrine/paracrine role for the IGF-I system in fallopian tube function, and their cyclic dependency further implies ovarian steroidal regulation.

Cellular localization of endothelial alkaline phosphatase reaction product and enzyme protein in the myocardium.

Myocardial capillary endothelial cells, arteriolar endothelial cells, and the arterial adventitia show positive alkaline phosphatase (AP) enzyme reaction and immunoreactivity in both rat and human hearts. In guinea pigs, however, capillary endothelial staining is discontinuous and arterial adventitia is negative. The ultrastructural correlate of discontinuous capillary staining is a pronounced labeling of pericytes in guinea pig heart and relatively weak endothelial staining. In rat and human heart, enzyme reaction products are localized mainly on plasma membranes and cytotic vesicles of endothelial cells. Comparison of two strains of rat reveals a more dense deposition of enzyme reaction product along the luminal and particularly along the abluminal plasma membrane of Sprague-Dawley rats than of Wistar rats. Quantitative analysis of immunogold labeled anti-AP antibody density confirms the pronounced polarity of capillary endothelial cell labeling in Sprague-Dawley rats. More than 80% of total endothelial AP protein in Sprague-Dawley rats is localized over the abluminal plasma membrane and basal lamina, as compared with less than 30% in Wistar rats. Moreover, the total endothelial cell labeling is almost sixfold higher in Sprague-Dawley than in Wistar rats. Total endothelial labeling and proportion of labeling on the abluminal endothelial plasma membrane in human hearts is intermediate between the two strains of rat. The strain and species differences in enzyme distribution could provide important information concerning enzyme function.

Immunocytochemical localization of prostaglandin endoperoxide synthase in the bovine intestine.

The localization of prostaglandin (PG) endoperoxide synthase in bovine intestine was examined immunocytochemically with polyclonal antibody raised against PG endoperoxide synthase purified from bovine seminal glands. The most intense positive staining reaction for the enzyme was present in mast cells. Mast cells were found to be widely distributed in the intestinal wall, and were particularly numerous in the lamina propria. Most of the mast cells in the lamina propria of the intestinal villi were elongated and oriented with their long axis parallel to the plane of the absorptive epithelium. In whole mount preparations of jejunal villi, mast cells were seen to form a two-dimensional network in the lamina propria. In addition to mast cells, smooth muscle cells of the inner circular muscle layer and muscularis mucosae, nerve cells and fibers, endothelial cells of arterioles, and serosal epithelial cells also showed faint to moderate staining for the enzyme. These results suggested that mast cells are the major source of PGs in the bovine intestinal wall. The characteristic arrangement of mast cells in the intestinal villi may be related to their functions in this portion of the bovine intestine.

IMPLICATIONS OF DE NOVO ELAM-1 AND VCAM-1 EXPRESSION IN HUMAN CARDIAC ALLOGRAFT REJECTION

Both cytokines produced by activated monocytes and T cells and direct cell-to-cell contact with antigen-primed T cells during inflammatory reactions are known to induce the expression of several adhesion proteins on endothelial cells. In this prospective longitudinal study, we analyzed the expression of ELAM-1, VCAM-1, and ICAM-1 on myocardial allograft biopsy specimens taken from 16 cardiac allograft recipients either for routine monitoring or for the investigation of suspected rejection. Infiltrating T cells were identified using anti-CD3 antibodies. Three to six sequential biopsies taken at one-week intervals were analyzed by means of conventional histology and immunohistochemistry. Seven patients did not develop rejection during the study; their biopsies were negative for VCAM-1 and ELAM-1, although faint ICAM-1 staining was present on capillaries, reflecting constitutive expression. Three patients entered the study with clear-cut clinical and histologic signs of acute rejection. Intense VCAM-1 and ICAM-1 expression was detected on capillary and postcapillary venules, together with a heavy CD3+ T cell infiltrate; VCAM-1 was also expressed on arteriolar endothelial cells. ELAM-1 was undetectable in all three cases. Six patients developed acute rejection during the course of the study. In four, ELAM-1 and VCAM-1 were expressed on both capillary and postcapillary venules one or two weeks before the histological diagnosis of rejection (heavy CD3+ cell infiltrate). Importantly, ELAM-1 expression was short-lived and had disappeared by the time CD3+ cellular infiltrate was detected, thus extending in vivo the finding that ELAM-1 expression is usually transient in vitro. Only VCAM-1 expression was observed in the other two patients, one week prior to the histological diagnosis of rejection. These results suggest that ELAM-1 and VCAM-1 might represent early predictive markers of acute cardiac allograft rejection. ELAM-1 expression is, however, usually transient, necessitating frequent testing.

A postembedding method using urea-unmasked ultrathin sections for the electron microscopic detection of sulfated glycoconjugates.

Using urea-unmasked ultrathin sections, a postembedding staining method in combination with nitrous acid procedure has been established for the efficient visualization of sulfated glycoconjugates in a series of ultrastructures of three salivary glands of the rat. Pieces of the parotid, submandibular and sublingual glands from adult rats were subjected to glutaraldehyde-paraformaldehyde fixation, rinsed, dehydrated and embedded in LR-White resin. Ultrathin sections were prepared, grid-mounted, incubated in urea (6M) aqueous solution, and then stained with an alcian blue (AB) pH 1.0-phosphotungstic acid (PTA) sequence with or without prior treatment with nitrous acid. The results obtained have shown that sulfated glycoconjugates are involved in such ultrastructures as mast cell granules, basal lamina of arteriolar endothelial cells and intercellular matrix of arteriolar smooth muscle cells.

Intracellular recording and dye transfer in arterioles during blood flow control.

We tested for dye coupling between arteriolar smooth muscle cells (SMC) and endothelial cells (EC) and investigated the correspondence of vasomotor activity with changes in the membrane potential (Vm) of EC and SMC during blood flow control. Female golden hamsters (n = 8, 90-170 g) were anesthetized (pentobarbital sodium, 60 mg/kg ip). A cheek pouch was spread over an optical pedestal, transilluminated, and irrigated with physiological saline solution (37 degrees C, pH 7.4). Glass microelectrodes were filled with 3 M KCl or with Lucifer yellow dye (LY, mol wt 470; 106 mM). SMC or EC of arterioles (ID, 20-50 microns) containing blood flow were impaled under a stereomicroscope. Vm was similar [-48 +/- 3 and -52 +/- 4 mV (means +/- SE)] with KCl (n = 6) or LY (n = 13) microelectrodes, respectively. Acetylcholine (5 x 10(-6) M) increased Vm from -47 +/- 3 to -67 +/- 4 mV (n = 5; P less than 0.01) concomitant with vasodilation. Spontaneous slow waves in Vm (2/min, 15-30 mV) were observed in arterioles with vasomotion. In cells identified with LY microinjection, resting Vm was -52 +/- 8 and -44 +/- 2 mV for EC (n = 3) and SMC (n = 3), respectively. SMC injected with LY did not show evidence of dye transfer to other SMC or to EC. When an EC was injected, the dye spread to many contiguous EC but not to SMC.(ABSTRACT TRUNCATED AT 250 WORDS)

モルモット肺傷害に及ぼすＬｉｐｏｓｏｍｅ化 Ｒｅｃｏｍｂｉｎａｎｔ ｈｕｍａｎ ｓｕｐｅｒｏｘｉｄｅ ｄｉｓｍｕｔａｓｅの影響

We investigated the effect of liposome-r-h-SOD on the lung injury induced by Forssman antiserum in guinea pigs. Intravenous injection of Forssman antiserum produced a biphasic increase in pulmonary resistance in guinea pigs. Liposome-r-h-SOD blocked this biphasic increase in a dose-dependent manner. Exudate and hemorrhage in the alveolar spaces and connective tissues were blocked by liposome-r-h-SOD. Liposome-r-h-SOD was more effective when it was injected 30 min prior to challenge than 5 min prior to challenge. On the other hand, free r-h-SOD or a mixture of free r-h-SOD and empty liposome did not block the lung injury. SOD activity in porcine endothelial cells cultured with liposome-r-h-SOD increased in a dose-dependent and time-dependent manner, while it did not increase with free r-h-SOD. Exogenous SOD was detected by immunoperoxidase staining in endothelial cells of arterioles and capillaries and in alveolar epithelial cells of the lung of guinea pigs injected with liposome-r-h-SOD. These findings suggest that superoxide radical may take part in the lung injury induced by Forssman antiserum. Liposome-r-h-SOD, which adheres to and/or is endocytosed by endothelial cells, may protect the lung from oxygen radicals.

The Limbal Vascular Response to Corneal Injury: An Autoradiographic Study

Thermal cautery of the peripheral cornea in rats caused proliferation of the limbal vasculature and invasion of the cornea. Tritiated-thymidine was used to identify premitotic activity in a total of 53,192 limbal vascular cells in five categories, viz., arteriolar endothelial cells, venular endothelial cells, arteriolar perivascular cells, venular perivascular cells, and capillary cells. From normal values in the range of 0.29 to 1.37%, the 2 h labeling indices reached a maximum of 13 to 14% in both endothelial and perivascular cells of venules and capillaries. Of particular interest was the finding of 18% labeling in arteriolar perivascular cells, and 7% in arteriolar endothelial cells. The categories showed a staggered onset of DNA synthesis, ranging from 17 h postcautery for capillary cells to 36 h for arteriolar endothelium and both arteriolar and venular perivascular cells. The duration of increased DNA synthesis also varied. Endothelial cells of both arterioles and venules showed narrow labeling peaks (12 to 24 h), while the adjacent perivascular cells and cells of the small vessels labeled for some 60 to 70 h. These results suggest that more than one stimulus to angiogenesis may be involved, or that the various cell types respond differently to the same stimulus.

Binding of 125I-epidermal growth factor in human uterus.

Quantitative light-microscopic autoradiography was used on five human uteri at two different phases of the menstrual cycle to ascertain the cell types with binding sites for epidermal growth factor (EGF). The results revealed that stromal cells, glandular epithelium of endometrium, elongated and circular muscle cells of myometrium, smooth muscle and endothelial cells of arterioles in the basal endometrium and myometrium contained numerous silver grains following incubation with 125I-EGF. Coincubation with 100-fold excess unlabeled EGF resulted in a complete disappearance of silver grains from all cell types. Quantitative grain analysis indicated that stromal cells contained the highest number of EGF-binding sites (P less than 0.05) with no significant differences among the others (P greater than 0.05). There was no cyclic variation of EGF-binding to any of the uterine cell types. The present data demonstrate that all the cell types of human uterus, including arterioles, contain EGF-binding sites. This suggests that all the cells in human uterus subserving different functions are targets of EGF action.