Photoactivated gamma-secretase inhibitors directed to the active site covalently label presenilin 1

Cleavage of amyloid precursor protein (APP) by the beta- and gamma-secretases generates the amino and carboxy termini, respectively, of the A beta amyloidogenic peptides A beta40 and A beta42--the major constituents of the amyloid plaques in the brain parenchyma of Alzheimer's disease patients. There is evidence that the polytopic membrane-spanning proteins, presenilin 1 and 2 (PS1 and PS2), are important determinants of gamma-secretase activity: mutations in PS1 and PS2 that are associated with early-onset familial Alzheimer's disease increase the production of A beta42 (refs 4-6), the more amyloidogenic peptide; gamma-secretase activity is reduced in neuronal cultures derived from PS1-deficient mouse embryos; and directed mutagenesis of two conserved aspartates in transmembrane segments of PS1 inactivates the ability of gamma-secretase to catalyse processing of APP within its transmembrane domain. It is unknown, however, whether PS1 (which has little or no homology to any known aspartyl protease) is itself a transmembrane aspartyl protease or a gamma-secretase cofactor, or helps to colocalize gamma-secretase and APP. Here we report photoaffinity labelling of PS1 (and PS2) by potent gamma-secretase inhibitors that were designed to function as transition state analogue inhibitors directed to the active site of an aspartyl protease. This observation indicates that PS1 (and PS2) may contain the active site of gamma-secretase. Interestingly, the intact, single-chain form of wild-type PS1 is not labelled by an active-site-directed photoaffinity probe, suggesting that intact wild-type PS1 may be an aspartyl protease zymogen.     

Frequency of the various stages of the seminiferous epithelium in different strains of male mice

Summary The duration of the various development stages of the seminiferous epithelium in different strains of male mice was determined by scoring the frequency distribution in randomly selected tubule cross-sections. The results obtained show a difference in the duration of the various stages between the different strains.Acknowledgments. The authors would like to thank Dr.J. Baarli and many other members of the Health Physics Group for their help and valuable assistance during the process of this work.     

Water and electrolyte movements across the duodenal mucosa in laying hens during in vivo perfusion]

The net water movements through the duodenum in laying hens are directly dependent on the applied osmotic gradient. The passive water permeability (Lp) of the mucosa is 6 times higher during absorption (mucosa to serosa: 70 microliter/h/mOsm) than in the opposite direction (secretion: 11.5 microliter/h/mOsm) and its semi permeability is 0.49 (reflexion coefficient sigma). Net sodium and potassium movements seem to be linked to net water flux. Duodenal water absorption should not be explained by a process of passive permeability.     

Effects of X-irradiation during hibernation on tissue catecholamine contents

Zusammenfassung 20 h nach Bestrahlung mit letaler Röntgenstrahlendosis (2000 r) zeigten winterschlafendeCitellus tridecemlineatus Herabsetzung der Konzentration von Catecholaminen in Leber und Niere. Werden Veränderungen des Wassergehaltes im Gewebe mitberücksichtigt, so erscheint der Gehalt an myokardialen Catecholaminen unverändert.

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Acknowledgments. This research was aided by Grants HE-06312, USPHS; and Ns G-271-62, NASA. The authors are grateful to DoctorD. E. Smith, Division of Biological and Medical Research, Argonne National Laboratory, for providing the ground squirrels and the facilities for their hibernation and radiation.     

Role of acoustic stress on the development of polyoma tumors in immunized golden hamsters]

An acoustic stress increases the metastatic spread of polyoma virus-induced tumors in syrian Hamsters. To try to elucidate the mechanism of this action, we studied the effect of stress on the protection afforded by the virus against tumour challenge. Exposure to ultrasonics during the immunization period markedly decreased the protective effect of the virus.     

Deafness and renal tubular acidosis in mice lacking the K-Cl co-transporter Kcc4

Hearing depends on a high K(+) concentration bathing the apical membranes of sensory hair cells. K(+) that has entered hair cells through apical mechanosensitive channels is transported to the stria vascularis for re-secretion into the scala media(). K(+) probably exits outer hair cells by KCNQ4 K(+) channels(), and is then transported by means of a gap junction system connecting supporting Deiters' cells and fibrocytes() back to the stria vascularis. We show here that mice lacking the K(+)/Cl(-) (K-Cl) co-transporter Kcc4 (coded for by Slc12a7) are deaf because their hair cells degenerate rapidly after the beginning of hearing. In the mature organ of Corti, Kcc4 is restricted to supporting cells of outer and inner hair cells. Our data suggest that Kcc4 is important for K(+) recycling() by siphoning K(+) ions after their exit from outer hair cells into supporting Deiters' cells, where K(+) enters the gap junction pathway. Similar to some human genetic syndromes(), deafness in Kcc4-deficient mice is associated with renal tubular acidosis. It probably results from an impairment of Cl(-) recycling across the basolateral membrane of acid-secreting alpha-intercalated cells of the distal nephron.