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Biomedical subjects

G Horn

Publications and source records attributed to G Horn.

At least 37 records · Page 2Linked to original sources
PubMed1997

Purification of ribonucleases Sa, Sa2, and Sa3 after expression in Escherichia coli.

The genes for three small ribonucleases from different strains of Streptomyces aureofaciens have been cloned and expressed in Escherichia coli. The purification of these ribonucleases from the periplasmic space is described. The yields range from 10 to 50 mg of protein per liter of culture medium. The molar absorption coefficients, isoelectric pH values, and pH of optimum activity are reported.

Amino Acid Sequence↗
PubMed1997

Delayed induction of a filial predisposition in the chick after anaesthesia.

In domestic chicks, Gallus gallus domesticus, early filial preferences are formed as the result of at least two processes: learning by exposure to conspicuous objects (filial imprinting) and a developing predisposition to approach some stimuli rather than others. The predisposition may be measured in visually naive chicks, e.g. as a preference for a rotating stuffed jungle fowl hen rather than for a rotating red box. Non-specific experience, such as handling or being placed in a running wheel in darkness, for a short time, is sufficient to induce the predisposition. In Experiment 1, the existence of a sensitive period for the induction of the predisposition, between approximately 14 and 42 h after hatching, was confirmed. The putative effect of anaesthesia on the induction of the predisposition was investigated in Experiment 2. Soon after hatching dark-reared chicks received injections of the anaesthetic equithesin (0.12 ml, I.P.), or saline, or they did not receive injections (controls). The chicks were subsequently placed in running wheels for a total of 2 h at either 24 h or 48 h after hatching. The following day the chicks' preferences were tested. In the 24 h-chicks, the saline and control groups showed a significant mean preference for the stuffed fowl, whilst the equithesin group did not. In contrast, in the 48 h-groups of chicks, only the equithesin group showed a significant mean preference for the stuffed fowl. These results suggest that the anaesthetic equithesin affects a developing filial predisposition in the domestic chick, and does so by delaying the onset of a sensitive period for the induction of the predisposition.

Anesthesia↗
PubMed1997

Clathrin proteins and recognition memory.

Strong converging evidence indicates that the intermediate and medial part of the hyperstriatum ventrale (IMHV) of the chick forebrain is a site of recognition memory for the learning process of imprinting. Clathrin proteins have been implicated in synaptic plasticity. In the present study we demonstrate for the first time that they are involved in vertebrate learning. Chicks were trained by exposure to a conspicuous object and their preference for it versus a novel object subsequently measured as a preference score (an index of learning). Trained chicks with low preference scores were classed as "poor learners" and those with high preference scores as "good learners". An additional group of chicks was untrained ("dark-reared"). Tissue was removed from the left and right IMHV, hyperstriatum accessorium and posterior neostriatum 9.5 h or 24 h after training. Clathrin heavy chain and clathrin light chains a and b were assayed using sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting. In the IMHV, and only for clathrin heavy chain, was there a significant effect of training. The effect occurred 24 h but not 9.5 h after training, and was significant only in the left IMHV. In this region at 24 h, there was (i) significantly more clathrin heavy chain in good learners than in dark-reared chicks, and (ii) a significant positive correlation between the amount of clathrin heavy chain and preference score; the amount of protein present in the dark-reared chicks did not differ significantly from the amount predicted from the regression line for trained chicks performing at chance (preference score 50). These findings imply that for the left IMHV, visual experience per se, locomotor activity and other side effects of training did not affect the amount of clathrin heavy chain. Rather, the increase observed was a function of the amounts chick learned and, because it was delayed, is likely to be involved in long-term memory. The results for clathrin heavy chain taken together suggest that enhanced presynaptic events in the IMHV, possibly associated with an increase in synaptic vesicle release/uptake, are important in the recognition memory underlying imprinting.

Animals↗
PubMed1997

[The use of the internal tissue expansion procedure in reconstructive surgery. Preliminary study and report of 2 cases].

The Frechet extender, initially proposed and successfully used in scalp reductions, can have many other indications in reconstructive surgery. This paper describes the preliminary results obtained after using this internal tissue extender in the treatment of a limb burn scar and a congenital giant naevus of the back. After a 3 month follow-up, these results are excellent. This procedure gives good results especially when there is a bony support underneath the skin to be treated.

Adolescent↗
PubMed1996

Differential interaction of the ras family GTP-binding proteins H-Ras, Rap1A, and R-Ras with the putative effector molecules Raf kinase and Ral-guanine nucleotide exchange factor.

The interactions of H-Ras, R-Ras, and Rap1A with the Ras-binding domains (RBD) of the c-Raf kinase and of the Ral guanine nucleotide exchange factor (RGF) was studied biochemically in solution. From deletion cloning the RGF-RBD was defined as a 97-amino acid-long fragment from the C-terminal end of the human RGF, which is an independent folding domain with high stability. Interestingly, whereas H-Ras binds with high affinity (KD = 20 nM) to Raf-RBD and with low affinity (KD = 1 microM) to RGF-RBD, Rap1A shows the opposite behavior. The binding of both RBDs to R-Ras is weak and shows no specificity. The interaction between Rap1A and RGF-RBD shows similar characteristics to the Ras-Raf interaction because it is blocked by mutations in the effector region (D38A) and it inhibits the dissociation of guanine nucleotide, which is the basis for the quantitative measurements in this work. Furthermore, the binding of RGF-RBD inhibits the interaction between Rap1A and Rap-GAP. As long as the cellular localizations of the different proteins and their biological functions are not clarified, these biochemical data seem to indicate that Ral-guanine nucleotide exchange factors is an effector molecule of Rap1A rather than of H-Ras.

Base Sequence↗
PubMed1996

Ras/Rap effector specificity determined by charge reversal.

Members of the Ras subfamily of small GTP-binding proteins have been shown to be promiscuous towards a variety of putative effector molecules such as the protein kinase c-Raf and the Ral-specific guanine nucleotide exchange factor (Ral-GEF). To address the question of specificity of interactions we have introduced the mutations E30D and K31E into Rap and show biochemically, by X-ray structure analysis and by transfection in vivo that the identical core effector region of Ras and Rap (residues 32-40) is responsible for molecular recognition, but that residues outside this region are responsible for the specificity of the interaction. The major determinant for the switch in specificity is the opposite charge of residue 31--Lys in Rap, Glu in Ras--which creates a favourable complementary interface for the Ras-Raf interaction.

Amino Acid Sequence↗
PubMed1995

Molecular cloning and functional analysis of the rat mu opioid receptor gene promoter.

The rat mu opioid receptor gene promoter was cloned and characterized. It has a few features in common with the mouse gene, e.g. the lack of a classical TATA box and the fact that several transcriptional start sites are used. The overall homology between the two species is greater than 85%. Functional analysis of the promoter was performed using transient expression of rat mu opioid receptor-reporter gene constructs in the mu opioid receptor expressing cell line SH SY5Y and in non mu opioid receptor expressing cell lines. A promoter region was defined which confers both high basal and TPA and forskolin stimulated reporter gene expression in SH SY5Y cells.

Animals↗
PubMed1995

The 2.2 A crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with Rap1A and a GTP analogue.

The X-ray crystal structure of the complex between the Ras-related protein Rap1A in the GTP-analogue (GppNHp) form and the Ras-binding domain (RBD) of the Ras effector molecule c-Raf1, a Ser/Thr-specific protein kinase, has been solved to a resolution of 2.2 A. It shows that RBD has the ubiquitin superfold and that the structure of Rap1A is very similar to that of Ras. The interaction between the two proteins is mediated by an apparent central antiparallel beta-sheet formed by strands B1-B2 from RBD and strands beta 2-beta 3 from Rap1A. Complex formation is mediated by main-chain and side-chain interactions of the so-called effector residues in the switch I region of Rap1A.

Amino Acid Sequence↗
PubMed1995

Differential distribution of protein kinase C (PKC alpha beta and PKC gamma) isoenzyme immunoreactivity in the chick brain.

Protein kinase C (PKC) is involved in neural plasticity. The phosphorylation of the myristoylated alanine-rich protein kinase C substrate (MARCKS) in the left intermediate and medial hyperstriatum ventrale (IMHV) of the chick brain has been shown previously to correlate significantly with the strength of learning in filial imprinting. The distribution of PKC alpha, beta I, beta II and PKC gamma in the brain of 1-day-old dark-reared chicks was determined immunocytochemically, using the monoclonal antibodies MC5 and 36G9, raised against purified PKC alpha beta and PKC gamma, respectively. PKC gamma-stained cells were distributed widely in the telencephalon, including all hyperstriatal structures (including the IMHV), the hippocampus, neostriatum, ectostriatum and archistriatum. There were fewer stained cells in the septum and the least cellular staining was in the paleostriatum primitivum. Fluorescent double-labelling with neuron-specific enolase (NSE) and with the glial calcium-binding protein S100 suggested that PKC gamma immunoreactivity was present in neurones but not in glia. The distribution of PKC alpha beta-stained cells was more limited, with staining in the archistriatum, hippocampus and septum but not in the hyperstriatum. However, there was PKC alpha beta-staining of some fibres in the IMHV (but little elsewhere in the hyperstriatum ventrale), in the neostriatum, paleostriatal complex and the lobus parolfactorius. Double-labelling with NSE and S100 revealed PKC alpha beta/S100-positive glial cells present in the paleostriatal region only. There was some PKC alpha beta-staining of putative neurones in the hippocampus, septum and archistriatum. The differential distribution of PKC isoenzymes suggests that in the IMHV some axonal inputs contain PKC alpha beta whereas some postsynaptic cells contain the gamma form of PKC.

Animals↗
PubMed1995

Neurophysiological investigations of a recognition memory system for imprinting in the domestic chick.

The responsiveness of neurons in a region of the chick brain involved in the learning process of imprinting, the right intermediate and medial hyperstriatum ventrale (right IMHV), has been investigated in unanaesthetized, trained and untrained chicks. The results demonstrate that neuronal responsiveness in this region reflects a variety of behavioural consequences of imprinting and is markedly altered as a result of the learning process. Groups of chicks (nine in each group) were either dark-reared or trained (imprinted) by exposure to a rotating red box or a rotating blue cylinder. Recordings of single or small groups of neurons were subsequently made from 156 sites in the right IMHV while the 2-day-old chicks were free to move in a running wheel. There was a highly significant increase in the proportion of sites responsive to the stimulus used to train the birds compared to the proportion responsive to that stimulus in dark-reared birds (30 and 9% respectively). These changes were found when either the red box or the blue cylinder was used to train the bird, the changes being similar for both stimuli. There was also a significant increase in the mean magnitude of the change in neuronal activity on stimulus presentation for the training stimulus compared to the same stimulus when not used in the bird's training. No significant effects of the training experience of the chicks were found upon either the magnitude of evoked activity or the proportion of sites responsive to a rotating stuffed jungle fowl or the sound of the maternal call. The presence of the training stimulus was selectively signalled by the response at certain sites. At other sites there was response generalization across stimulus shape or colour. A comparison with results for the left IMHV demonstrates both similarities and differences in neuronal responsiveness between the two regions. In both regions imprinting selectively enhances neuronal responsiveness to the training stimulus. However, for trained birds the mean proportion of sites responding to whichever of the red box or the blue cylinder was not used in the bird's training was significantly lower in the right than the left IMHV. These results are discussed in relationship to previously reported asymmetries in the response of the right and left IMHV regions to imprinting. A model is introduced to explain the physiological findings. The effects of training on right IMHV neuronal function are consistent with a long-term role for this region in the recognition memory of imprinting.

Acoustic Stimulation↗
PubMed1995

The expression of proenkephalin and prodynorphin genes and the induction of c-fos gene by dopaminergic drugs are not altered in the straitum of MPTP-treated mice.

The expression of proenkephalin (PENK), prodynorphin (PDYN) and c-fos genes was studied in the striatum of C57B1/6 mice treated with 1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine (MPTP), which are used as a rodent model of Parkinson's disease (PD). Two weeks after systemic administration of MPTP (2 x 40 mg/kg, s.c. 18h apart), the lesion of the substantia nigra (SN) could be visualised by loss of the nigral tyrosine hydroxylase (TH) mRNA hybridization signal and by a 91% decrease in striatal dopamine levels. The levels of PENK and PDYN mRNAs were not significantly changed in the striatum of the lesioned mice, as compared to non-treated controls. The induction of the immediate early gene c-fos by the dopamine D2 receptor antagonist haloperidol was not altered, while the selective D1 receptor agonist SKF 38393 failed to induce c-fos in the striatum of MPTP-treated mice. These results are in contrast to the data concerning rats with the 6-hydroxydopamine (6-OHDA) lesion of the SN, which serve as another rodent model of PD. In the striata of 6-OHDA-lesioned rats, PENK gene is upregulated, PDYN gene is down-regulated and the induction of c-fos gene by D2 receptor antagonists is abolished, whereas selective D1 receptor agonists induce c-fos gene, which does not occur in non-lesioned rats. We presume that the lack of influence of the MPTP lesion in mice on the striatal gene expression was mainly caused by insufficient dopamine depletion in the striatum, which could not be increased in this model. The importance of the changes observed in 6-OHDA-lesioned rats has been discussed in the context of the mouse and primate MPTP models of PD.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1995

Functionally distinct memories for imprinting stimuli: behavioral and neural dissociations.

Precocial chicks exposed to a stimulus subsequently approach that stimulus in preference to other, novel stimuli. Previous investigations of the neural basis of these imprinting preferences suggest that imprinting training results in the formation of two memories. The first memory is formed rapidly and is located in the intermediate and medial part of the hyperstriatum ventrale (IMHV) of the left hemisphere; the formation of the second, in another memory system, S', takes several hours and can be prevented by a lesion placed in the right IMHV soon after training. The results of the present study suggest that the functional characteristics of these memories differ. Although memories in both left IMHV and S' supported imprinting preferences (Experiments 1a and 2a), only memories in S' influenced the acquisition of a heat-reinforced discrimination in which imprinted objects served as discriminanda (Experiments 1b and 2b).

Animals↗
PubMed1994

Learning-related changes in Fos-like immunoreactivity in the chick forebrain after imprinting.

The intermediate and medial part of the hyperstriatum ventrale (IMHV) is a part of the chick forebrain that is critical for the learning process of imprinting and may be a site of information storage. Chicks were either trained on an imprinting stimulus or dark-reared. Trained chicks were classified as good or poor learners by their preference score (a measure of the strength of imprinting). A monoclonal antibody against the immediate early gene product Fos was applied to sections through IMHV and other forebrain regions. In the IMHV, significantly more immunopositive nuclei were counted in good learners than in poor learners or dark-reared chicks. There was a positive correlation between counts of labeled nuclei and preference score that was not attributable to sensory activity per se, locomotor activity during training, or a predisposition to learn well; rather, the results indicated that the change in Fos immunoreactivity in the IMHV was related to learning. In the hyperstriatum accessorium, significantly fewer immunopositive nuclei were counted in good learners than in poor learners or in dark-reared chicks. In the dorsolateral hippocampal region, more immunopositive nuclei were counted in trained than in dark-reared chicks. No significant effects of training were found in the anterior hyperstriatum ventrale, lobus parolfactorius, neostriatum, medial hippocampal region, or ventrolateral hippocampal region, but counts in this last region were positively correlated with training approach. The results for IMHV implicate Fos or Fos-related proteins in memory processes and pave the way for the identification of the cell types that show the learning-related increase in gene expression.

Animals↗
PubMed1994

Learning-related alterations in the visual responsiveness of neurons in a memory system of the chick brain.

The intermediate and medial hyperstriatum ventrale (IMHV) of the chick brain is known to be essential for the learning process of imprinting. The activity of neurons was recorded from the left IMHV of 2-day-old unanaesthetized chicks while the chicks were free to move in a running wheel. The chicks were either raised in complete darkness or visually trained (imprinted) with a set duration of exposure to a visual image. The first group of these birds was trained by exposure for 100 min to a rotating red box and the second was trained by similar exposure to a rotating blue cylinder. A third group was left untrained. Training more than doubled the proportion of sites that responded to the stimulus used to train the bird, relative to the proportion of sites responsive to the other stimulus and to the proportion of sites responsive in the untrained birds; the learning-related increase was selective and highly significant. Behavioural monitoring indicated that the enhanced responsiveness could not be explained by overt differences in the alertness, attentiveness or movements of the birds. No significant effect of training was found on the proportion of sites responsive to a rotating stuffed jungle fowl or to the sound of a maternal call. The response at certain sites selectively signalled the presence of the training stimulus, while at others the response showed generalization across stimulus shape or colour. There was a non-specific effect of training upon the pattern of spontaneous discharges of the neurons: the numbers of spikes occurring in clusters (bursts) was significantly reduced in trained birds compared with the dark reared controls.

Acoustic Stimulation↗
PubMed1994

NADPH diaphorase (nitric oxide synthase) in a part of the chick brain involved in imprinting.

The intermediate and medial part of the hyperstriatum ventrale (IMHV) is a memory system in the chick forebrain in which certain learning-related changes occur after imprinting. We have enquired whether NADPH-diaphorase, a presumed marker for nitric oxide synthase, is present in sections through the IMHV of chicks, either trained with an imprinting stimulus or dark-reared. Very few NADPH-d-positive cells were found in the IMHV (0.37 +/- 0.07 S.E.M. cells per 0.3 mm x 0.9 mm sampling frame), in contrast to the palaeostriatum augmentatum (PA) (19.47 +/- 0.77). Some stained cells in the PA were closely associated with blood vessels. The results do not support the hypothesis that learning-related changes in the IMHV depend on nitric oxide acting as a retrograde neuronal messenger.

Amino Acid Oxidoreductases↗
PubMed1994

mRNA distribution of two isoforms of somatostatin receptor 2 (mSSTR2A and mSSTR2B) in mouse brain.

The primary gene transcript of the mouse somatostatin receptor 2 is alternatively spliced giving rise to two isoforms (mSSTR2A and mSSTR2B) which differ at the C-terminus. Using reverse transcription polymerase chain reaction (RT-PCR), both mRNAs were found in the cortex, hippocampus, hypothalamus, striatum, mesencephalon, cerebellum, medulla oblongata, pituitary and in testis, however with different ratios between mSSTR2A and mSSTR2B, implicating a tissue-specific control of transcription and splicing. Among the analyzed tissues, cortex contained the highest amounts of mSSTR2A but only little mSSTR2B, whereas the pons/medulla oblongata expressed both isoforms to an equal extent. Northern blot analysis of these tissues revealed a single mRNA of about 2.4 kb using a mSSTR2A-specific hybridization probe. No additional signal was seen using a probe which hybridizes to both mSSTR2A and mSSTR2B, suggesting that the two mRNAs may be nearly identical in length. In addition, in situ hybridization indicated that mSSTR2A is predominantly expressed in mouse brain, and mSSTR2B is never expressed independently from mSSTR2A.

Animals↗
PubMed1994

The role of stimulus comparison in perceptual learning: an investigation with the domestic chick.

In two experiments an imprinting procedure was used to familiarize chicks with two stimuli, A and B, that subsequently served as the discriminanda in a simultaneous discrimination. On the first day of each experiment, subjects either received presentations of A and B that were intermixed within a session (mixed exposure) or presentations of A in one session and of B in another (separate exposure). For half of the subjects in each of the exposure conditions, A and B differed in both colour and form; for the remainder A and B differed in form alone. On the second day of the experiments, the chicks were placed into a cool test apparatus and given training in which approaching A was rewarded by the delivery of a stream of warm air, but approaching B was not. Acquisition of this discrimination was more rapid when A and B differed in two respects than when they differed in form alone. When A and B differed in both colour and form, the heat-reinforced discrimination was acquired more rapidly after separate exposure than after mixed exposure; but when A and B differed in form alone, discrimination learning was more rapid following mixed exposure than separate exposure. The latter finding, that the opportunity to compare stimuli differing in only one dimension facilitates subsequent discrimination learning, is consistent with earlier suggestions (Gibson, 1969) regarding the conditions that promote perceptual learning.

Animals↗