Immunocytochemical demonstration of calmodulin in cells secreting by exocytosis

Calmodulin is a regulator of several calcium-dependent cellular processes. It has been suggested that it plays a role in the mechanism of secretion. Employing an indirect immunoperoxidase technique at the light microscope level, this study demonstrates the presence of calmodulin in several exocytotic cells (mast cells, thyroid follicular cells, neurohypophyseal neurosecretory terminals, pancreatic beta-cells and pancreatic acinus cells) in rat and man. The positive staining reaction for calmodulin was granular and at least in the case of rat mast cells it appeared to be associated with the granule membrane.     

Immunocytochemical demonstration of contractile cells in the human ovarian follicle

Summary Actin-and myosin-like immunoreactivity is found in cells located in the theca externa of the follicle wall of the human ovary, and corresponding to previously observed myoid cells. The immunocytochemical observation provides direct structural evidence that non-vascular contractile cells are also present in the follicle wall in humans. As expected, perifollicular blood vessels showed a positive immunoreaction for actin and myosin in their smooth muscle walls.This work was supported by the Swedish Medical Research Council, grant No. 14X-732/5680.     

Immunocytochemical demonstration of contractile cells in the human ovarian follicle

Actin- and myosin-like immunoreactivity is found in cells located in the theca externa of the follicle wall of the human ovary, and corresponding to previously observed myoid cells. The immunocytochemical observation provides direct structural evidence that non-vascular contractile cells are also present in the follicle wall in humans. As expected, perifollicular blood vessels showed a positive immunoreaction for actin and myosin in their smooth muscle walls.     

Possible involvement of calmodulin in apical constriction of neuroepithelial cells and elevation of neural folds in the chick

Summary Chlorpromazine and trifluoperazine HCl, antipsychotic drugs known to bind to calmodulin, reversibly inhibited elevation of neural folds by interfering with the contractile activity of apical microfilament bundles in developing chick neuroepithelial cells.We wish to thank C. Hess for her technical assistance. This study was supported in part by grants from the Busch Fund (Rutgers University), UMDNJ Foundation and Kapnek Fund.     

Effect of the calmodulin inhibitor R24571 (calmidazolium) on rat embryos cultured in vitro

Summary The possible effects of inhibition of the calcium-binding protein, calmodulin, on mammalian morphogenesis have been investigated by culturing rat embryos in vitro from 9 1/2 to 11 1/2 days of development in the presence of R24571 (calmidazolium), a specific inhibitor of calmoldulin. Embryos cultured in 10^–2 mM R24571 for 48 h show inhibited development and exhibit a range of morphogenetic abnormalities including assymetry and neural tube defects. Embryos exposed to R24571 for the first 24 h of a 48 h culture are more severely affected than embryos exposed to R24571 for the last 24 h.Acknowledgments. We wish to thank Mr C. Veltkamp and Mr B. lewis for their expert help with the scanning electron microscopy and photography.     

Enhancement of the cytogenetic efficacy of the antitumor agent bleomycin by the calcium and calmodulin antagonist fendiline

Summary The induction of chromosome aberrations (dicentric and ring chromosomes) in human lymphocytes by the antitumor agent bleomycin is synergistically enhanced when bleomycin is applied together with the calcium antagonist fendiline (Sensit^®).     

Contraction of myoepithelial cells in secreting submandibular glands of dogs

Summary In slowly secreting submandibular glands of dogs, a sympathetic effect on myoepithelial cells was demonstrated as an acceleration of the flow, follwed by retardation, on short-lasting sympathetic stimulation, particularly after -adrenoceptor-blocking drugs.Supported by grant No. 14X-00539 from the Swedish Medical Research Council     

In situ demonstration of the activity of 3 beta-hydroxysteroid dehydrogenase on steroid hormone secreting cells within the paraaortic lymph nodes of golden hamster

The in situ activity of 3 beta-hydroxysteroid dehydrogenase was detected in clustered cells which showed steroid-producing morphology, within the capsule of the paraaortic lymph node. In light and electron microscopic studies, the positive reaction products were detected on intracapsular cell clusters. This result indicates that these unique cells may have a steroid secreting function within the lymph nodes.     

Contraction of myoepithelial cells in secreting submandibular glands of dogs.

In slowly secreting submandibular glands of dogs, a sympathetic effect on myoepithelial cells was demonstrated as an acceleration of the flow, followed by retardation, on short-lasting sympathetic stimulation, particularly after beta-adrenoceptor-blocking drugs.     

Cytosolic free Ca2+ in insulin secreting cells and its regulation by isolated organelles

The role of Ca2+ in secretagogue-induced insulin release is documented not only by the measurements of 45Ca fluxes in pancreatic islets, but also, by direct monitoring of cytosolic free Ca2+, [Ca2+]i. As demonstrated, using the fluorescent indicator quin 2, glyceraldehyde, carbamylcholine and alanine raise [Ca2+]i in the insulin secreting cell line RINm5F, whereas glucose has a similar effect in pancreatic islet cells. The regulation of cellular Ca2+ homeostasis by organelles from a rat insulinoma, was investigated with a Ca2+ selective electrode. The results suggest that both the endoplasmic reticulum and the mitochondria participate in this regulation, albeit at different Ca2+ concentrations. By contrast, the secretory granules do not appear to be involved in the short-term regulation of [Ca2+]i. Evidence is presented that inositol 1,4,5-trisphosphate, which is shown to mobilize Ca2+ from the endoplasmic reticulum, is acting as an intracellular mediator in the stimulation of insulin release.     

Cytosolic free Ca2+ in insulin secreting cells and its regulation by isolated organelles

Summary The role of Ca²⁺ in secretagogue-induced insulin release is documented not only by the measurements of⁴⁵Ca fluxes in pancreatic islets, but also, by direct monitoring of cytosolic free Ca²⁺, [Ca²⁺]_i. As demonstrated, using the fluorescent indicator quin 2, glyceraldehyde, carbamylcholine and alanine raise [Ca²⁺]_i in the insulin secreting cell line RINm5F, whereas glucose has a similar effect in pancreatic islet cells. The regulation of cellular Ca²⁺ homeostasis by organelles from a rat insulinoma, was investigated with a Ca²⁺ selective electrode. The results suggest that both the endoplasmic reticulum and the mitochondria participate in this regulation, albeit at different Ca²⁺ concentrations. By contrast, the secretory granules do not appear to be involved in the short-term regulation of [Ca²⁺]_i. Evidence is presented that inositol 1,4,5-trisphosphate, which is shown to mobilize Ca²⁺ from the endoplasmic reticulum, is acting as an intracellular mediator in the stimulation of insulin release.     

Histochemical demonstration of carbonic anhydrase activity in mast cells

Zusammenfassung Carboanhydraseaktivität in Mastzellen der Ratte wurde mittels Modifikation der Methode vonHäusler ⁴ demonstriert. Kapillarendothel, Basalzellen des Zungenepithels und Muskelgewebe reagierten ebenfalls positiv.     

Metallothionein induction by cadmium and zinc in rat secretory organs

Summary Metallothionein (MT) levels were determined in four secretory organs of the rat following administration of zinc (Zn) and cadmium (Cd). The concentrations of MT in the lacrimal, parotid and adrenal glands of untreated rats were in the range of 2.2–4.9 g/g wet weight tissue while in the pancreas it was shown to be 15.2 g/g. Injection of zinc at total doses of 16, 32 and 80 mg/kg resulted in a 1.8-, 3.2- and 5.9-fold increase in lacrimal MT content, respectively, while a 10.2- and 13.1-fold elevation was observed following treatment with 4 and 8 mg/kg of Cd, respectively. Similar findings were found in the adrenal gland. The parotid MT was elevated 5.9 and 17 times following Zn treatment at doses of 16 and 80 mg/kg respectively, whereas 4 mg/kg of Cd increased MT 14.4 times in this gland. Pancreatic MT was elevated by 39- and 40-fold after injection of Zn at doses of 16 and 32 mg/kg respectively, whereas 4 and 8 mg/kg of Cd caused a 9.8- and 17.9-fold induction, respectively. These results may indicate that secretory organs participate in metabolism of heavy metals in the mammalian body.     

Loss of antibody production accompanied by chromosome loss in a cloned hybrid line secreting antibodies to sheep red blood cells

Summary Somatic cell hybrids between Sp2/O-Ag14 mouse myeloma cells and lymphocytes derived from BALB/c mice hyperimmunized with sheep red blood cells (SRBC) were produced. One hybrid producing IgG₁ antibody to SRBC was selected, cloned twice and subsequently transferred to BALB/c mice. After a number of transfers it was found that the antibody titer in ascitec fluid gradually decreased. Cytogenetic analysis revealed gradual chromosome loss in the hybrid clone, which produced progressively less antibody.     

Loss of antibody production accompanied by chromosome loss in a cloned hybrid line secreting antibodies to sheep red blood cells

Somatic cell hybrids between Sp2/O-Ag14 mouse myeloma cells and lymphocytes derived from BALB/c mice hyperimmunized with sheep red blood cells (SRBC) were produced. One hybrid producing IgG1 antibody to SRBC was selected, cloned twice and subsequently transferred to BALB/c mice. After a number of transfers it was found that the antibody titer in ascites fluid gradually decreased. Cytogenetic analysis revealed gradual chromosome loss in the hybrid clone, which produced progressively less antibody.     

Relationship between lectin-affinity granules in anuran embryos and formation of primordial germ cells

Summary Embryos of the anuran,Rana nigromaculata, contained granules with a specific affinity for Con A and GS-I. Larvae derived from embryos injected with these lectins had a noticeable reduction in both the number and size of primordial germ cells (PGCs). This observation suggests that the granules are somehow involved in the formation of PGCs.     

Identification of hemolytic granules isolated from human myocardial cells

Summary Human myocardial cells from fresh autopsy material contained granules which possessed hemolytic activity against guinea pig and rabbit erythrocytes. The hemolytic granules, which had a density of 1.02 and a diameter of 200–300 nm, were recovered as a microsome fraction from subcellular homogenates of human myocardial cells by differential centrifugation in 300 mM sucrose containing 0.1 mM PMSF and 10 mM EDTA. The membrane lesions caused by the granules were ring-like structures with an internal diameter of about 10–17 nm, analogous to that caused by perforin- and complement-induced lysis. However, the requirement for divalent cation differed from that for perforin-induced lysis, since the microsome-mediated lysis occurred in the presence of EDTA.