The roles of autophosphorylation and phosphorylation in the life of osteopontin

Osteopotin is a secreted glycosylated phosphoprotein found in bone and other normal and malignant tissues. Osteopontin can be autophosphorylated on tyrosine residues and can also be phosphorylated on serine and threonine residues by several protein kinases. Autophosphorylation of osteopontin may generate sites for specific interactions with other proteins on the cell surface and/or within the extracelluar matrix. These interactions of osteopontin are thought to be essential for bone mineralization and function. The polyaspartic acid motif of osteopontin, in combination with neighboring sequences that include serine residues phosphorylated by protein kinases, could fold and assemble into a molecular structure that participates in the mineralization of the bone matrix.     

Quantitative autoradiographic localization of neuropeptide Y (NPY) binding sites in rat posterior pituitary lobe

1. We have localized and quantified neuropeptide Y (NPY) binding sites in the rat pituitary gland after incubation of tissue sections in the presence of 125I-Bolton-Hunter NPY followed by autoradiography, computerized microdensitometry, and comparison to 125I-standards. 2. In the rat, NPY binding sites are localized exclusively to the part of the posterior pituitary lobe closer to the pituitary stalk. No NPY binding sites could be found in the intermediate or the anterior pituitary lobes. 3. Our results suggest a role for NPY in the regulation of pituitary function and, in particular, that of the neural lobe.     

Allocation of Photosynthate to Individual Tubers of Solanum tuberosum L.: I. RELATIONSHIP BETWEEN TUBER GROWTH RATE AND ENZYME ACTIVITIES OF THE STARCH METABOLISM

Potato plants (Solanum tuberosum L.) were grown in water culturein a controlled environment. Cooling (+8°C) of individualtubers decreased their growth rates and increased the growthrates of non-cooled tubers of the same plant. The carbohydrateconcentration in non-cooled and cooled tubers did not differsignificantly, but ¹⁴C-import from labelled photosynthate waslower in cooled than in non-cooled tubers. The markedly lowerconversion rate of ethanol-soluble ¹⁴C to starch in cooled,in comparison to non-cooled tubers, was not associated withsignificant differences in the in vitro activities of starchsynthase, ADPG-pyrophosphorylase and starch phosphorylase understandard assay conditions (+30°C). However, the Q₁₀-valuesof the enzymes differed in vitro in the temperature range between30°C and 8°C, leading to a marked decrease in the activityratio of ADPG-pyrophosphorylase/starch phosphorylase in cooledtubers. In tubers differing in growth rates without manipulation, 14d after tuber initiation significant positive correlations werefound between ¹⁴C-concentration of tuber tissue and the in vitroactivities of starch synthase and ADPG-pyrophosphorylase anda significant negative correlation between ¹⁴C-concentrationand starch phosphorylase. In contrast, in tubers which wereanalysed 5 d after initiation, there were only small differencesbetween tubers in growth rate, ¹⁴C import and the activity ratioADPG-pyrophosphorylase/starch phosphorylase. From various directand indirect evidence it is concluded that the growth rate ofindividual tubers, and thus the sink strength, is at least inpart controlled by the activity of starch synthesizing enzymes. Key words: Potato tuber, cooling, starch synthesizing enzymes     

Peripheral benzodiazepine binding sites in kidney: modifications by diabetes insipidus

Using [3H] diazepam as ligand, it is possible to distinguish neuronal binding sites from those present on glial elements and in peripheral tissues (non-neuronal). The function of the "non-neuronal" binding sites is still obscure. Preliminary data showed a distribution of [3H] diazepam binding sites in kidney that could suggest a localization along the renal tubules. This is the site at which a renal peptide, arginine-vasopressin (AVP) is supposed to act. In an attempt to examine the function of these "non-neuronal" sites, we studied the [3H] diazepam binding in kidney of Brattleboro rats which lack AVP and present the symptoms of diabetes insipidus. The homozygous Brattleboro rats showed an increase in the apparent number of benzodiazepine binding sites (Bmax) compared to Long-Evans control rats. Replacement of AVP in these animals results in a reversal of the electrolyte alterations of diabetes insipidus and in an increase of the affinity of the [3H] diazepam binding. These findings may indicate a possible relationship between benzodiazepine binding sites and vasopressin action in kidney and may support receptor function of these "non-neuronal" binding sites.     

Increased atrial natriuretic peptide (6-33) binding sites in the subfornical organ of water deprived and Brattleboro rats

Binding sites for rat atrial natriuretic peptide (6-33) (ANP) were quantitated in the subfornical organ of chronically dehydrated homozygous Brattleboro rats unable to synthesize vasopressin; heterozygous Brattleboro rats, their controls, Long Evans rats and Long Evans rats after 4 days of water deprivation. Brain sections were incubated in the presence of 125I-ANP and the results analyzed by autoradiography coupled to computerized microdensitometry and comparison to 125I-standards. Brattleboro rats and water deprived Long Evans rats presented a higher number of ANP binding sites than their normally hydrated controls. Our results suggest a role of ANP binding sites in the subfornical organ in the central regulation of fluid balance and vasopressin secretion.     

Localization of atrial natriuretic peptide, ANP-(99-126) binding sites in the rat thymus and spleen with quantitative autoradiography

Binding sites for atrial natriuretic peptide, ANP-(99-126) were studied in lymphoid organs of the rat with quantitative autoradiography. Tissue sections were incubated in the presence of 0.13 nM 125I-ANP-(99-126) followed by autoradiography using [3H]-Ultrofilm, and the results were analyzed by computerized densitometry and comparison to 125I-standards. Specific ANP binding sites were localized in the medulla and the cortex of the rat thymus and in the white pulp of the rat spleen, with apparent binding sites concentrations of 93, 65, and 126 fmol/mg protein, respectively. The presence of ANP binding sites in areas related to the maturation and function of lymphocytes, and to the production of thymic hormones, suggests the possibility of a role of circulating ANP in the modulation of the immune response.     

Degeneration in the parvocellular portion of the lateral geniculate nucleus of the cebus monkey

Summary The synaptology of the Cebus lateral geniculate nucleus (LGN) was studied after varying (3–15 days) periods of survival following unilateral and bilateral eye enucleations. Part of the material was processed with the Glees and Nauta techniques for light microscopy while the rest was processed for electron microscope observation. The study revealed a variety of degenerated terminals in the parvocellular portion of the LGN and allowed the differentiation of the retinal from the extraretinal terminals. The most frequent synaptic type of retinal origin is a glomerular large central terminal (up to 20 long) which makes axodendritic and axoaxonic synaptic contacts with geniculate dendrites and peripheral small terminals. Simple axodendritic and axosomatic terminals of retinal and extraretinal origin were also found. The early changes affecting the geniculate neurons and astrocytes during the degenerative process are described.These results are discussed in relation to: 1) previous work on the LGN synaptology of cats and macaques; 2) the physiology of the LGN; 3) the phagocytic role of astrocytes; 4) the general problem of degeneration in the central nervous system. In addition, a correlation between the light and electron microscope observations is attempted.Work supported by Grants from the National Council to Combat Blindness, Inc. N.Y., U.S.A. (Fight for Sight Grant-in-Aid-G-340), the AF-AFOSR Grant No. 963/67-68, and the Consejo Nacional de Investigaciones Cientificas y Técnicas, Buenos Aires, Argentina.The authors acknowledge the continuous advice and encouragement received from Prof. E. de Robertis throughout all the phases of the project. The expert technical assistance of Miss E. di Matteo, Mr. A. Sáenz and Mr. R. Castelli is also gratefully acknowledged.     

Further studies on the understanding of Octodon degus natural resistance to morphine: a comparative study with the Wistar rat

1. Octodon degus shows higher levels of tolerance to morphine when compared with the Wistar rat. 2. In the formalin algesiometric test, this caviomorph is more resistant to pain (P less than 0.01) and to the analgesic effect of morphine (P less than 0.001). 3. CD50 and LD50 were significantly higher in Octodon degus as compared with Wistar rat. 4. Morphine caused in rat severe hypotension, while doses eight times higher in O. degus had a transient effect. 5. 3H-naloxone binding in adrenal glands of O. degus is higher than in other tissue samples assayed from the same animal or rats.