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[Functional differentiation of different portions of the the rat neocortex].

The effect of uni- and bilateral ablation of rostral and caudal parts of neocortex on conditioned avoidance reflex was studied on naive and pretrained albino rats. Maximal impairment of the reflex was seen after bilateral ablation of the caudal region of neocortex, minimal one--after unilateral neocortex ablation. The rats without rostral cortex showed medium impairment. After extirpation of caudal cortex the number of positive reactions to light was greater in pretrained rats than in naive ones. After total cortical extirpation the previously elaborated conditioned reflex disappeared irreversibly. The conclusion is made on functional heterogeneity of the rostral and caudal cortex.

Animals

Functional differentiation of the chick endocrine pancreas. II. The alpha cells and glucagon.

A radioimmunoassay for glucagon, together with electron microscopic observations of early embryonic alpha cells were utilized to examine developmental aspects of glucagon accumulation and release in the chick embryo. Immunoreactive glucagon was detected in both the pancreas and blood plasma from the fifth embryonic day onwards. In addition, emiocytotic events were observed in alpha cells as early as the fifth embryonic day. The early appearance of glucagon and its subsequent developmental profile correlate well with major events in carbohydrate metabolism occurring in the embryonic chick, and are discussed in relation to a functionally responding system, the developing liver. The present data show that glucagon is secreted at earlier embryonic stages than hitherto demonstrated, and suggest a developmental role for glucagon in hepatic glycogen metabolism.

Animals

Distinct cell morphotypes of Aureobasidium melanogenum ZN exhibit differential functional profiles in promoting maize growth.

Black yeast-like fungi of the genus Aureobasidium exhibit morphological plasticity, but whether distinct cellular states within the same genetic background are associated with different plant growth-promoting functions remains unclear. Here, yeast-like cells (YL), swollen cells (SC), and chlamydospores (CH) of Aureobasidium melanogenum ZN were characterized. YL was associated mainly with siderophore production and laccase activity, SC with extracellular polysaccharide accumulation, and CH with phosphate mobilization and higher ammonia and IAA production. Whole-genome and comparative genomic analyses revealed a shared repertoire related to nutrient acquisition, auxin-associated metabolism, extracellular oxidation, and carbohydrate remodeling, with expansions in nutrient- and cell-surface-related gene families. Transcriptomic and metabolomic analyses showed distinct deployment of these capacities, with CH exhibiting broad reprogramming of tryptophan-associated, nitrogen, phosphate, central-carbon, and amino-acid metabolism. In maize, CH at the optimal inoculation concentration of 105 CFU·mL-1 produced the strongest growth promotion, increasing plant height, dry biomass, root length, root surface area, and root volume by 58.6%, 365.1%, 191.0%, 194.3%, and 222.4%, respectively. Consistent with this pronounced growth phenotype, maize root transcriptomics showed coordinated CH-induced responses involving root development, nutrient transport, redox regulation, and root-interface remodeling. Root-zone tracking showed greater short-term stability and persistence of CH. These findings identify cellular state as an important functional dimension of Aureobasidium-plant interactions and provide a basis for developing fungal inoculants with defined beneficial cellular states.

Zea mays

Differential function of major histocompatibility complex antigens in T-lymphocyte activation.

We have emphasised the functional dichotomy of MHC LD and LD antigens as well as the differences in cellular responses to these antigens. Perhaps in so doing we have failed to stress adequately the similarities that exist. But while the similarities (for example skin graft rejection associated with both K and I region differences) are so very clear, the differences have best allowed our progressive understanding of MHC induced cellular responses from the perspective stressed in this article. Of greatest importance to our understanding of these transplantation antigens are the potentially differential roles for the LD and SD antigens in the complex series of events that are collectively referred to as the "allograft reaction". It has been suggested that these differences may be "merely quantitative". This possibility has been discussed repeatedly in our previous reports on the distinction of LD and SD. In fact, the great bulk of biological phenomena can be reduced to quantitative differences. It would seem to us that sufficient evidence for such differential activity exists to make the LD-SD dichotomy model an heuristically valuable one for purposes of designing future experiments. We have discussed the clinical relevance of this model elsewhere. Many authors have speculated and evidence has been gathered to suggest, that cell surface antigens associated with the MHC are important in developmental and other cell interactions. Some studies have directly addressed the question of the need for MHC compatibility to allow cell interaction to proceed optimally. It thus seems most appropriate that the genetic complex with which we are dealing has been termed the major histocompatibility complex; allowing for the literal interpretation of this term this may be the genetic region that by its influence on "tissue compatibility" may control critical cellular interactions in addition to those observed in allograft reactions. It is the simple good fortune for those whose attention was focused on this complex by transplantation problems to find themselves with a panorama of biological phenomena that require extensive experimental probing and integration, hopefully ultimately leading to an understanding of the MHC in a broader context than has to date been possible.

Animals

Functional differentiation of splanchnic A delta fibres in relation to viscerosomatic reflexes.

The splanchnic afferent signal in the myelinated fibre spectrum was analysed in cats and its relationship to viscerosomatic reflex activity was studied. In addition to Abeta and Adelta components, a further elevation was observed in the sympathetic chain neurogram. According to the conduction rate (15 plus or minus 4 m/sec), it was caused by myelinated fibres 2-3 mu in diameter. The presence of this component at thoracic dorsal root level, or in the sympathetic chain in stimulation of the postganglionic part of the nerve trunk, confirmed the afferent nature of the observed activity. The threshold stimulus for these fibres, which were termed group Adelta2, was four times higher than for Abeta fibres. Correlation of the neurogram with evoked activity in the intercostal nerves confirmed the different functional role of group Adelta2 fibres. Adelta2 fibre activity evokes the early component of the two-component viscerosomatic discharge and Adelta2 fibre activity its later component. The authors discuss the functional significance of Adelta2 fibres, which probably mediate nociceptive information from the viscera.

Animals

REgional functional differentiation in the gut of the grasscarp, Ctenopharyngodon idella (Val.).

A regional differentiation--reflecting structural differences--of the intestine of larval and juvenile grasscarps can be illustrated by studying the activity of alkaline phosphatase and the uptake of orally administered horseradish peroxidase. Pinocytosis takes place in a welldefined area of about 23% of the length of the gut (segment II). Neither the rostral +/- 68% (segment I) nor the caudal +/- 9% (segment III) shows absorption of the enzyme. Alkaline phosphatase activity, mainly localized at the microvilli of the enterocytes is high in the first segment of the gut and low in the second segment. In larvae, the activity decreases sharply at the transition from segment I to segment II. The activity is weak or absent in the caudal third segment. Quantitative histochemical data are confirmed by biochemical analyses. Alkaline phosphatase activity is found all over the mucosal folds of the first segment, with relatively weak activity at the base and at the tip of the folds. This may be related to a renewal of the epithelium. Our results suggest that active absorption of digested food takes place mainly in the rostral first segment, while the uptake of macromolecules by pinocytosis is a function of the second segment. Comparison of the results with information available in literature leads to a rejection of the hypothesis that the uptake of protein macromolecules in Cyprinids is to be attributed to the absence of a stomach and therefore to an inefficient digestion of proteins.

Aging

Abnormal neuronal differentiation (functional maturation) in mental retardation.

A review is presented of a variety of human pathologic conditions, including some forms of mental retardation, and of experimental situations involving the nervous system proved (with the Golgi method) to be associated with detectable morphologic abnormalities. The postsynaptic elements (the dendritic spines) have been found to be especially sensitive and frequently they are found to be structurally abnormal. Structural abnormalities involving one of the synaptic components could result in synaptic dysfunction which could explain some degree of mental or motor retardation or incoordination. It is hoped that this review will stimulate and encourage the use of the Golgi method in the study of abnormal conditions (clinical or experimental) affecting the nervous system.

Cell Differentiation

Studies on the persistence of differentiated functions in rat hepatocytes set into primary tissue culture. II. Production of specific exportable proteins and the effect of purine cyclic nucleotides: an immunofluorescent study.

Immunofluorescent studies showed that even after 15 days in vitro primary neonatal rat hepatocytes contained in their cytoplasm detectable amounts of different adult rat serum proteins, including fibrinogen and proalbumin. Estimation of the intensity of specific fluorescence revealed that in untreated cultures the hepatocytic content of the various exportable antigens progressively diminished between the 5th and 15th day in vitro. Treatment with cAMP (10(-5) M daily) alone increased in hepatocytic cytoplasm, with respect to parallel controls, the content of total exportable proteins and of proalbumin. Daily administration of an equimolar association (10(-5) M) of cAMP with cGMP increased the total protein, proalbumin and fibrinogen content of hepatocytes. Daily treatment with cGMP (10(-5) M) alone caused only light and transitory increases in the content of proalbumin and fibrinogen. Rocket immune electrophoresis showed that the hepatocytic secretion of specific proteins into the growth medium persisted up to the 15th day, although progressively diminishing in intensity. The secretion of total exportable proteins and of albumin, but not of fibrinogen, was stimulated by cGMP used alone or coupled with equimolar cAMP.

Cell Differentiation

Evidence for differential function of neuronal and glial cells in protein metabolism and amino acid transport.

Amino acid incorporation in neuronal and glial cells has been investigated in several laboratories employing bulk-separation techniques to obtain cell-enriched fractions. The relative rates of incorporation into the proteins of both cell types vary substantially with the method of isotope administration. Through the use of single-pulse perfusions with a duration of 30-40 sec the early time course of labeling has been studied. The difference between neuronal and glial cells with respect to indicating cell interactions will be discussed. The in vitro amino acid incorporation has been measured in neurons and glia after slice incubation. In material from animals developing experimental allergic encephalitis the rate of 3H-leucine incorporation more than doubles in the unfractionated brain. Glial cells increased their rate of incorporation by approximately 400% under the same conditions. The involvement of specific proteins in the cells and organelles has been studied by gel electrophoresis. The high uptake capacity of glial cells for certain amino acids with possible transmitter function has been further characterized. The release of these substances is measured in a superfusion system where beds of cells, preloaded with the labeled substance, are used. High potassium pulses stimulate release of, for example, GABA in both neuronal and glial cells.

Amino Acids

Differential function of the phosphoglucomutase isozymes PGM1 and PGM2.

A total of 13 metabolites thought to be possibly inhibitory were tested for their influence on PGM isozyme activities, each at several different concentrations. The analysis of statistical significance was based on enzyme activities obtained by densitometric measurements of starch gels. Five of the substances were found to inhibit PGM activity, three of which definitely and a further one probably led to a significantly stronger inhibition of the isozymes of the PGM2 locus than of PGM1 isozymes. They are (1) fructose-1,6-diphosphate, (2) adenosine triphosphate, (3) citrate, and (4) possibly 2,3-diphosphoglycerate. Thus, PGM1 isozymes proved to function better in hard or perhaps marginal metabolic conditions. Related evolutionary aspects are discussed.

Adenosine Triphosphate

Expression of differentiated functions in hepatoma cell hybrids: IX extinction and reexpression of liver-specific enzymes in rat hepatoma-Chinese hamster fibroblast hybrids.

Most of the hybrid clones derived from a cross of Chinese hamster fibroblasts (DON) with rat hepatoma cells (Faza 967) showed preferential loss of rat chromosomes. Two of the hybrid clones retained the rat chromosomes, and both showed extinction of 4 liver-specific enzymes: aldolase B, liver alcohol dehydrogenase, and the inducible enzymes tyrosine aminotransferase and alanine aminotransferase. Subcloning of 1 of these hybrids, which contained 2 sets of hepatoma chromosomes and 1 set of hamster chromosomes, permitted the isolation of some clones which reexpressed 1 or more of the liver-specific enzymes. Liver alcohol dehydrogenase was the most frequently reexpressed enzyme and aldolase B the least. Tyrosine aminotransferase inducibility was reexpressed independently of basal activity, and the enzyme produced by the reexpressing hybrid cells was precipitated by a specific antiserum. No correlation was detected between the presence or absence of the marker chromosomes (large metacentrics) of the hamster parent and the extinction and reexpression of the hepatic enzymes. The results reported confirm and extend to interspecific hybrids the observation of the stable and independent reexpression of tissue-specific enzymes.

Alanine Transaminase

Expression of differentiated functions in hepatoma cell hybrids: selection in glucose-free media of segregated hybrid cells which reexpress gluconeogenic enzymes.

Selective glucose-free media have been used to study the reexpression of liver-specific gluconeogenic enzymes in rat hepatoma X mouse lymphoblastoma somatic hybrids. The utilization for gluconeogenesis of dihydroxyacetone or oxaloacetate requires two enzymes: fructose diphosphatase as well as either triokinase for the former or phosphoenolpyruvate carboxykinase for the latter. By sequential selection with these substrates, the reexpression of the three gluconeogenic enzymes has been dissociated. The reexpression of these enzymes is correlated with the loss of mouse chromosomes. In addition, the characterization of the parental forms of aldolase B, another liver-specific enzyme, shows that reexpression corresponds to the simultaneous production of the rat and mouse enzymes. These results demonstrate the chromosomal origin of extinction and suggest that activation of mouse silent genes which accompanies reexpression can occur without loss of the parental determinations. The hypothesis that determination involves regulatory rather than structural genes is discussed.

Alcohol Oxidoreductases

Primary monolayer cultures of postnatal rat liver cells with extended differentiated functions.

Monolayers of liver cells cultured from postnatal rats were grown in two types of media. One set of cultures was grown in selective medium which contained ornithine but was deficient in arginine; the other set was grown in nonselective medium which contained arginine but no ornithine. The cultures that were grown in the nonselective medium contained primarily a mixture of two cell types found in the liver: parenchymal hepatocytes and fibroblast-like cells. The fibroblast cells tended to overgrow the hepatocytes after several days in culture. In contrast, fibroblast overgrowth was inhibited in cultures grown in the selective, arginine-deficient medium, thereby resulting in relatively pure cultures of functional parenchymal hepatocytes. Comparative studies of sulfobromophthalein (BSP) uptake showed that the cultures grown in selective medium continued to be active much longer than the cultures grown in the nonselective medium. Pyruvate kinase assays revealed that the cultures grown in selective medium contained primarily the L-isoenzyme type which is characteristic of parenchymal hepatocytes. Cultures grown in nonselective medium contained a mixture of L- and M-isoenzymes which is indicative of nonparenchymal liver cells. The reported results indicate that selective, arginine-deficient medium permits primarily the growth of parenchymal hepatocytes found in neonatal rat liver.

Animals

Morpho-functional differentiation in lobular carcinoma of the breast.

Lobular carcinoma of the breast has been studied using histochemical methods for mucosubstances; immunocytochemical methods for casein and actin; the ruthenium red electronycytochemical method for acid glycoproteins and an immunoelectroncytochemical method for casein. Mucosubstances and casein showed a similar cytoplasmic localization, but casein production was much more intense and also showed a more diffuse cytoplasmic localization. Occasionally casein assumed the form of target-like 'inclusions' as seen characteristically with the mucosubstances. The neoplastic cells were not stained by antisera against actin. Ultrastructurally, some cells showed an intracytoplasmic lumen with microvilli and/or an irregular outline at one extremity which was covered by microvilli. An electron-dense 'fuzz' and casein coated the microvilli of cells exposed respectively to ruthenium red and an anticasein serum followed by peroxidase--anti-peroxidase complexes. It is concluded that lobular carcinoma shows evidence of epithelial rather than myoepithelial differentiation with the emphasis on epithelial secretory cells engaged in intensive milk protein production. All 10 tumours tested for oestrogen receptors were positive in contradistinction to ductal carcinoma with a lower incidence of positivity. It appears that, in addition to distinctive histological and histochemical features, lobular carcinoma has an almost constant endocrine pattern in respect of its oestrogen receptor content.

Actins