M. Schachter

Effect of nerve stimulation, denervation, and duct ligation, on kallikrein content and duct cell granules of the cat's submandibular gland

Various procedures which reduce or deplete the kallikrein content of the cats submandibular gland correspondingly reduce the number of apical granules in the striated duct cells. The kallikrein content is greatly reduced after chronic parasympathetic but not after sympathetic nerve section which suggests that the parasympathetic innervation is required for synthesis or storage of this enzyme.

Purification and Partial Characterization of Cat Pancreatic and Urinary Kallikreins — Comparison with Other Cat Tissue Kallikreins and Related Proteases

Kallikreins have been purified from cat pancreas and urine by methods similar to those described previously for cat colon and submandibular gland kallikreins. The pancreatic kallikrein (M.W. 41,200, pI 4.75) was similar to the urinary kallikrein (M.W. 34,300 pI 4.35-4.70) in pH optimum, substrate specificity and inhibition profile. Both enzymes were potent kininogenases and immunologically similar. These enzymes closely resembled the kallikreins from cat colon and submandibular glands. A trypsin (M.W. 18,800) was isolated from cat pancreas and shown to be distinct from the group of kallikreins in all parameters tested. We attempted purification of cat renal kallikrein, but were unable to isolate any such enzyme. The major acidic esterase of cat kidney cortex (M.W. 59,000, pI 4.91) was purified and was distinct from both the cat tissue kallikreins and trypsin. The origin of cat urinary kallikrein remains unclear, but in the light of our findings, it may result from renal filtration of blood-borne tissue kallikreins rather than from intrarenal synthesis.

The Varied Localization and Functional Significance of Kallikrein-Like Enzymes in Salivary Glands, Pancreas, Colon, Sex Glands and Spermatozoa, Including Evidence for the Presence of Nerve Growth Factor (NGF) in Bull Sperm Acrosome

There has been considerable recent progress in the cellular and subcellular localization of kallikreins (or kininogenases) and related serine proteases.1–8 In this report, we review briefly our localization studies - some published and some new observations. These include studies on the salivary glands and colon of various mammals, the sex glands of the guinea pig, and the spermatozoa of boar, bull and man. In addition, we report our observations on the presence of Nerve Growth Factor (NGF) in bull sperm.

Purification of kallikrein from cat submaxillary gland

Cat submaxillary gland has been used in many investigations on kallikrein-kinin system. Though it has not yet been established, the physiological functions of kallikreins in various glands, such as the functional vasodilatation, have been studied mainly in cat submaxillary gland (1, 2, 3). Furthermore, the secretory cells or subcellular distribution has been studied in this gland (4). Meanwhile the isolation of submaxillary kallikrein has not yet been achieved. In a series of investigation on the proteases in the rat submaxillary gland, Ekfors, et al. obtained a kallikrein-like peptidase (5), but the identity of the enzyme with kallikrein was not clear because of the lack of study on the biological activity of the preparation. Fiedler, et al. described that hog submaxillary kallikrein was separated into a number of active components with isoelectric points in the range pH 3.3–4.4 (6), but the specific activity of their preparation was less than a half of that of purified hog pancreatic kallikrein. We attempted the isolation of kallikrein from the cat submaxillary gland and obtained a highly purified preparation which was homogeneous in disc electrophoresis.

Gastric secretion during anaesthesia and its inhibition by metiamide and other drugs.

the histamine H2-receptor antagonist, metiamide, inhibits the acid gastric secretion produced by chloralose-urethane anaesthesia in dogs carrying gastric cannulae chronically. This secretion is also prevented by atropine and hexamethonium. ‘Spontaneous’ gastric secretion of vagal origin in conscious dogs is also blocked by metiamide.

Purification and Partial Characterization of Cat Colon and Submandibular Gland Kallikreins

Kallikreins from cat colon and submandibular gland have been purified by acetone fractionation of tissue extracts, DEAE-Sephacel ion-exchange chromatography, rho-aminobenzamidine Sepharose 4B affinity chromatography and gel filtration on Sephadex G-75. They were of similar M.W., approximately 40,000, and each comprised five forms by isoelectric-focusing (pI 4.1-4.8). Both enzymes were potent kininogenases and exhibited similar specificities with synthetic ester and amide substrates. They were susceptible to a range of protease inhibitors. Surprisingly, neither was sensitive to aprotinin yet both were partially inhibited by soya-bean trypsin inhibitor. They were indistinguishable in our immunological tests. An acidic esterase (pI 2.2-3.5) of M.W. 120,000 was isolated from cat stomach by the same procedure. While it exhibited weak immunologic similarity to cat submandibular gland kallikrein, it had negligible kininogenase activity and different substrate and inhibitor specificities to the two kallikreins. It is concluded that similar tissue kallikreins are present in the colon and submandibular gland of the cat but are distinct from this cat stomach esterase.

Kininogenase in kidney after ligation of the ureter and after experimental aortic stenosis

Nachweis, dass Abbinden des Harnleiters von Ratten in 1–3 Wochen eine fortschreitende und ausgesprochene Verminderung der Kiniogenase in der Niere verursacht. Experimentelle Verengerung der Aorta kingegen hat entweder keine Wirkung auf die Nieren-Kininogenase oder erhöht sie nur geringfügig.

Sympathetic vasodilation in the submaxillary gland and its enhancement after chronic parasympathetic denervation

Die «Nacherweiterung» der Gefässe, welche auf Reizung des sympathischen Nervs der Submaxillaris-Drüse in der Katze folgt, wird nicht durch Kallikrein oder Kinin ausgelöst, da sie nicht durch den hochwirksamen synthetischen Bradykinin-potenzierenden Faktors (BPF) beeinflusst wird. Eine neue Beobachtung zeigte, dass diese sympathische Gefässerweiterung bedeutend verstärkt wird, nachdem der parasympathische Nerv durchschnitten wurde und degeneriert ist.

Recent studies on kallikrein in the submandibular gland of the cat

Kallikrein By a combination of methodologies, studies have been made to collect precise information about kallikrein in order to establish its physiological role. By correlating effects of nerve stimulation, degenerative nerve section, duct obstruction and other procedures with enzyme content of the glands and with ultrastructural changes using electron and immunofluorescent light microscopy, we have come to the following conclusions for the cats submandibular gland: (1) Kallikrein is located luminally, in the apical region of the cells of the striated ducts. It is absent from acinar, demilune and other cells, and is secreted into the duct system and not into the intelstitium of the gland. The correlation of enzyme content and microscopic appearance suggests that the enzyme is contained in small apical secretory granules of the striated ducts [1-3]. These results suggest a specific role for these ducts and for kallikrein in the salivary glands. (2) Kallikrein largely disappears from the cats submandibular gland 1-2 weeks after parasympathetic but not after sympathetic denervation. This is correlated with corresponding changes in kailikrein content of the gland and with the disappearance of apical granules of the striated ducts. Our results suggest that the synthesis or storage of kallikrein in these cells depends on an intact parasympathetic nerve supply. (3) The very specific location of kallikrein in striated duct cells suggests a specific role for this cell and for this enzyme. (4) The presence in the submandibular gland of known active macromolecules like kallikrein, renin, nerve growth factor, and of a new one like sialotonin [4] suggests that the physiological significance of this gland may extend beyond those relatively simple digestive functions generally assigned to it.