Left IMHV Research Articles

Light Color Changes The Density and Height of Synapses in an Area of the Brain Essential For Learning

The effects of pre and post-passive avoidance training, and exposure to different wavelengths of light on numerical synapse density and the mean projected synapse height in the intermediate and medial part of the hyperstriatum ventrale (IMHV) of the chick brain were investigated. Synaptic counts and the mean projected synapse height measurements were done using a 3-dimensional stereological probe, the disector, which requires no assumptions about size, shape, and orientation of synapses. It was found that there was a significant increase in the density of synapses in the left IMHV of chicks in white, yellow and red light illuminations respectively. However, the mean projected height of synapses in these chicks was significantly increased in the right IMHV under white, yellow, green but not red light illumination conditions. These results suggest that numerical and structural modification of synapses may depend on wavelengths of white, yellow, red and green lights.

Amino acid neurotransmitter release and learning: a study of visual imprinting

The intermediate and medial part of the hyperstriatum ventrale (IMHV) is an area of the domestic chick forebrain that stores information acquired through the learning process of imprinting. The effects of visual imprinting on the release of the amino acids aspartate, arginine, citrulline, γ-aminobutyric acid (GABA), glutamate, glycine and taurine from the left and right IMHVs in vitro were measured at 3.5, 10 and 24 h after training. Chicks were exposed to an imprinting stimulus for 1 h, their preferences measured 10 min afterward and a preference score calculated as a measure of the strength of learning. Potassium stimulation was used to evoke amino acid release from the IMHVs of trained and untrained chicks in the presence and absence of extracellular Ca 2+. Ca 2+-dependent, K +-evoked release of glutamate was significantly (34.4%) higher in trained than in untrained chicks. This effect was not influenced by time after training or by side (left or right IMHV). Training influenced the evoked release of GABA and taurine from the left IMHV at both 3.5 and 10 h. The training effects at the two times were statistically homogeneous so data (≤10 h group) were combined for each amino acid respectively. For this ≤10 h group, evoked release increased significantly with preference score. In contrast, for the 24 h group, evoked release of GABA and taurine was not significantly correlated with preference score. There were no significant correlations between preference score and GABA or taurine release in the right IMHV at any time, nor in the absence of extracellular calcium. No significant effects of training condition, time or side were observed for any other amino acid in the study. The present findings suggest that soon after chicks have been exposed to an imprinting stimulus glutamatergic excitatory transmission in IMHV is enhanced, and remains enhanced for at least 24 h. In contrast, the learning-related elevations in taurine and GABA release are not sustained over this period. The change in GABA release may reflect a transient increase in inhibitory transmission in the left IMHV.

Analysis of differential gene expression supports a role for amyloid precursor protein and a protein kinase C substrate (MARCKS) in long-term memory.

Previous work has identified the intermediate and medial part of the hyperstriatum ventrale (IMHV) as a region of the chick brain storing information acquired through the learning process of imprinting. We have examined in this brain region changes in expression of candidate genes involved in memory. Chicks were exposed to a rotating red box and the strength of their preference for it, a measure of learning, determined. Brain samples were removed approximately 24 h after training. Candidate genes whose expressions were different in IMHV samples derived from strongly imprinted chicks relative to those from chicks showing little or no learning were identified using subtractive hybridization. The translation products of two candidate genes were investigated further in samples from the left and right IMHV and from two other brain regions not previously implicated in imprinting, the left and right posterior neostriatum. One of the proteins was the amyloid precursor protein (APP), the other was myristoylated alanine rich C kinase substrate (MARCKS). In the left IMHV the levels of the two proteins increased with the strength of learning. The effects in the right IMHV were not significantly different from those in the left. There were no effects of learning in the posterior neostriatum. This is the first study to relate changes in the amounts of MARCKS and APP proteins to the strength of learning in a brain region known to be a memory store and demonstrates that the systematic identification of protein molecules involved in memory formation is possible.

Short communication: hippocampal neuronal activity and imprinting in the behaving domestic chick.

The hippocampus of the chick projects to the intermediate and medial part of the hyperstriatum ventrale (IMHV) which stores information acquired through the learning process of imprinting. We have investigated whether the response properties of hippocampal neurons are similar to those of IMHV neurons. Chicks were imprinted by exposure, one group (n = 7) to a rotating red box (RB), the other (n = 5) to a rotating blue cylinder (BC). Four chicks were untrained. The following day, when the chicks were approximately 48 h old, neuronal activity was recorded in the left hippocampus. The proportion of neurons responding to the RB and that to the BC in untrained chicks were compared with the proportions in trained birds. (i) In RB-trained chicks both the proportion responding to the RB and that to the BC were significantly increased. (ii) In BC-trained chicks no significant effect on these proportions was found. Of the responsive neurons some were colour (red or blue) sensitive and others were shape (box or cylinder) sensitive; the proportions so responsive were not influenced by training condition. Certain neurons responded significantly differently when a stimulus was 0.5 m or 2 m from the chick (35%; d-sensitive); very few neurons were equivalently responsive to a stimulus at both distances (3%; d-invariant). These proportions were not significantly affected by training condition. Hippocampal responses are compared with those in the left IMHV. It is concluded that IMHV responses do not passively reflect those of hippocampal neurons.

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.

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).

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.

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).

Effects of light hatching on synapse number and size in the intermediate and medial part of the hyperstriatum ventrale of the domestic chick

The effects of pre-hatch light exposure on synaptic development in the intermediate and medial part of the hyperstriatum ventrale (IMHV) of the chick brain were investigated. Quantitative electron microscopical techniques were used to determine the size and numerical density of synapses and it was seen that in light hatched chicks there was a significant increase in the density of synapses in the left IMHV but that the size of synapses in these birds was decreased. These results provide a link between synaptic development and plasticity.

Neuronal activity related to memory in the intermediate and medial part of the hyperstriatum ventrale of the chick brain

The intermediate and medial part of the hyperstriatum ventrale (IMHV) in the forebrain of the domestic chick Gallus gallus domesticus has been shown in previous studies to be critically involved in the learning process of imprinting. In the present study, 1-day-old chicks were imprinted by exposing them to one of two artificial stimuli. 24 h later each chick was given a preference test in which the two stimuli were presented in sequence. A preference score, an index of the strength of imprinting (i.e., of learning), was calculated from approach activity during the preference test. The chicks were divided into groups with low, medium and high preference scores (corresponding to weak, medium and strong learning respectively) and coded so that all subsequent procedures were performed blind. Each chick was then aneesthetized and spontaneous action potentials recorded extracellularly from groups of neurones in the left IMHV. The mean neuronal firing rate in chicks with medium and high preference scores was significantly greater than that in chicks with low preference scores. This relation between neuronal activity and preference score was not attributable to the chicks' locomotor activity per se. The results demonstrate an association between spontaneous electrical activity in the left IMHV and a measure of learning 1 day after the learning occurred.

Learning selectively increases protein kinase C substrate phosphorylation in specific regions of the chick brain.

The effect of imprinting, an early form of exposure learning, on the phosphorylation state of the protein kinase C substrates myristoylated alanine-rich C-kinase substrate (MARCKS) and protein F1/43-kDa growth-associated protein (F1/GAP-43) was studied in two regions of the chick forebrain. One region, the intermediate and medial part of the hyperstriatum ventrale (IMHV), is probably a site of long-term memory; the other, the wulst, contains somatic sensory and visual projection areas. After imprinting, a significant increase in MARCKS protein phosphorylation was observed in the left IMHV but not the right IMHV. No significant alteration in F1/GAP-43 was observed in IMHV. MARCKS was resolved into two acidic components of pI approximately 5.0 and approximately 4.0. Phosphorylation of the pI approximately 5.0 MARCKS but not the pI approximately 4.0 MARCKS was significantly altered by imprinting. The partial correlation between preference score (an index of learning) and phosphorylation, holding constant the effect of approach activity during training, was significant only for the pI approximately 5.0 MARCKS in the left IMHV. A significant negative partial correlation between preference score and F1/GAP-43 phosphorylation in the right wulst was observed. Because the imprinting-induced alteration in MARCKS is selective with respect to phosphoprotein moiety, hemispheric location, and brain region, we propose that these alterations may be central to the learning process.

Learning-dependent Changes in the Responses to Visual Stimuli of Neurons in a Recognition Memory System.

When young chicks are trained by exposing them to a conspicuous object they learn its characteristics. The learning process is known as imprinting. In the present study neuronal activity in a region crucial for imprinting was shown to be affected by training and by the object on which the chicks had been trained. The region is the intermediate and medial part of the left hyperstriatum ventrale (left IMHV). No such effects were found in a visual projection area, the left hyperstriatum accessorium. Domestic chicks were imprinted on either a rotating red box (n=7 chicks) or a rotating blue box (n=8). When the chicks were approximately 48 h old they were anaesthetized and multiple-unit activity was recorded in simultaneous, single penetrations through each of the two regions. Records were also made from eight dark-reared chicks. Whilst recording, the red or blue box, placed in front of the contralateral eye, was switched on to give a total of 20 rotations, the interval between each rotation being 10 s. The alternative stimulus was then presented 20 times. Unit activity in the 3 s before and after stimulus onset was compared and the data for each of the 20 presentations were combined. In the left IMHV 18 out of a total of 115 recording sites (16%) responded significantly to the stimuli; in the left hyperstriatum accessorium 39 out of 126 recording sites (26%) did so. Measures of unit activity at each recording site were combined for a given penetration to provide a 'mean penetration response'. The response to the red box differed from the response to the blue box in the left IMHV of dark-reared chicks. After training with the blue box the response to both boxes was similar to the response to the blue box in dark-reared birds. After training with the red box the response to both boxes was similar to the response to the red box in dark-reared birds. No significant effects were found in the left hyperstriatum accessorium. The two training boxes were virtually identical apart from the differences in colour and brightness. Training appeared to stabilize the response of the visually naive left IMHV to the training stimulus whilst changing its response to the alternative, but similar stimulus. That is, one consequence of training is that the two stimuli are placed in the same category, and this neural change may provide a basis for stimulus generalization. The underlying neural system is modelled and a mechanism that allows such stimuli to be discriminated is proposed.

Impairment of learning by localized injection of an N-methyl-D-aspartate receptor antagonist into the hyperstriatum ventrale of the domestic chick.

A restricted part of the domestic chick forebrain is critically involved in the learning process of imprinting. This region is the intermediate and medial part of the hyperstriatum ventrale (IMHV). The effect on imprinting of local injection of the N-methyl-D-aspartate (NMDA) receptor blocker D-amino-5-phosphonopentanoic acid (D-AP5) into the left IMHV was studied in chicks in which the right IMHV had been lesioned. The left IMHV is essential for imprinting when chicks have been lesioned in this way. Injection of approximately 0.7 nmol D-AP5 into the left IMHV significantly impaired imprinting. Injection of approximately 0.2 nmol D-AP5 into the left IMHV, or of approximately 0.7 nmol D-AP5 into the left hyperstriatum accessorium, was without significant effect on imprinting. These results suggest that NMDA receptors in the left IMHV may play an important part in this learning process.

The development of hemispheric asymmetries in neuronal activity in the domestic chick after visual experience

Previous experiments have shown that a restricted part of the forebrain, the intermediate and medial part of the hyperstriatum ventrale (IMHV), is involved in the learning process of imprinting. Through this process chicks selectively approach, and so recognise, a visually conspicuous object to which they have been exposed. Previous work has also demonstrated that the left and right IMHV regions play different roles in the recognition memory of imprinting, and that exposure to an object results in changes in synaptic organisation in the left, but not in the right IMHV. We have enquired whether training has a differential effect on spontaneous multiple-unit impulse activity in the two regions. Thirty-two chicks were reared in darkness. Sixteen were trained by exposing them to a flashing, rotating box; the remaining chicks serving as dark-reared controls. The trained chicks were anaesthetised either immediately after training (0-h trained chicks) or returned to the dark incubator and anaesthetised 6 h after the end of training (6-h trained chicks). A single microelectrode penetration was made through the left IMHV simultaneously with a single penetration through the right IMHV; a single penetration was also made through a visual projection area, the left hyperstriatum accessorium. Recordings were made from between 3 and 7 sites in each penetration, and the mean multiple-unit firing rate for these sites was calculated. Spontaneous activity in the right IMHV changed in respect of activity in the left IMHV with time after training; no such effects were found in the left hyperstriatum accessorium.(ABSTRACT TRUNCATED AT 250 WORDS)

Synaptic transmission and recognition memory: Time course of changes in N-methyl-D-aspartate receptors after imprinting.

Previous studies have shown that a restricted region of the chick forebrain (the intermediate and medial parts of the hyperstriatum ventrale, IMHV) is crucial for the learning process of imprinting. A learning-related increase in N-methyl-D-aspartate (NMDA) receptor binding has previously been shown in the left IMHV 8.5 hr after training with an imprinting stimulus. The present experiments showed that this increase in binding in the left IMHV does not occur until 6-8.5 hr after training: There was no significant change in binding in either the left or right IMHV less than 0.5, 3, or 6 hr after training. No significant change in binding occurred in the right IMHV 8.5 hr after training. It is probable that the increase in binding in the left IMHV is associated with the recognition memory of imprinting.

Memory systems in the chick: Dissociations and neuronal analysis

Young domestic chicks learn to recognize the visual characteristics of an object to which they are exposed. A restricted part of the forebrain, the intermediate and medial part of the hyperstriatum ventrale (IMHV) is implicated in this process. This form of exposure learning can be dissociated from (i) the ability to learn certain procedures or skills, and (ii) a predisposition to attend to particular types of naturalistic objects. The first of these dissociations is reminiscent of that found in certain human organic amnesias, whilst the second may have its counterpart in the processes involved in face recognition by infants. The right and left IMHV play different roles in the memory that underlies imprinting. The cellular and molecular processes involved in this form of memory are discussed.

Learning and memory: regional changes in N-methyl-D-aspartate receptors in the chick brain after imprinting.

An extensive series of experiments has implicated a restricted region of the chick forebrain in the learning process of imprinting. The region is the intermediate and medial part of the hyperstriatum ventrale (IMHV). Previous studies have shown that training is associated with an increase in the area of the postsynaptic density of axospinous synapses in the left but not the right IMHV. The postsynaptic density is a site of high receptor density, and at least some axospinous synapses are excitatory. We found that imprinting is associated with a 59% increase in N-methyl-D-aspartate-sensitive binding of the excitatory amino acid L-[3H]glutamic acid in the left IMHV. The increase is probably due to an increased number of binding sites. The profile of sensitivity of the sites to a series of amino-, phosphono-substituted carboxylic acids (2-amino-3-phosphonopropionate to 2-amino-8-phosphonooctanoate) is characteristic of N-methyl-D-aspartate-type receptors. There were no significant effects of training on binding in the right IMHV. The effect of training on left IMHV binding could not be attributed to light exposure, arousal, or motor activity per se but was a function of how much the chicks learned. The changes in the left IMHV could increase the effectiveness of synaptic transmission in a region crucial for information storage and so form a neural basis for recognition memory.

The effects of protein synthesis inhibition on structural changes associated with learning in the chick

The effect of the protein synthesis inhibitor anisomycin on the structural changes associated with passive avoidance learning in the chick was investigated. Chicks were trained when they were 24 h old by allowing them to peck at a shiny bead coated with either water or the aversive-tasting substance methylanthranilate (MeA). Chicks which peck the MeA-coated bead will on subsequent testing avoid pecking a similar, but water-coated bead. Behavioural testing was carried out 12 h after training and immediately afterwards the chicks were killed and their brains prepared for electron microscopy. A specific region of the forebrain, the intermediate and medial part of the hyperstriatum ventrale (IMHV) was investigated. When the IMHV of the MeA trained chicks was compared with that of water-trained controls structural changes of the synapse were detected. These changes involved a significant increase in the mean length of the postsynaptic density ( L PSD ) of symmetrical synapses in the left IMHV. Chicks injected with 0.8 mg of anisomycin 30 min before training with a MeA-coated bead showed aversion for the shiny bead when tested 12 h later. Electron microscopic analysis of the IMHV from these amnestic chicks showed no evidence for the change in L PSD demonstrated in the water-injected controls. These results are discussed in relation to the nature of the memory trace induced by training on a passive avoidance task.

Imprinting, learning, and memory.

If a visually naive chick is exposed to one of a wide range of conspicuous objects, the chick may learn its characteristics. A series of biochemical studies has implicated a restricted part of the forebrain in this process of imprinting; a specific region (IMHV) has been identified which may be a site of information storage. Changes in the morphology of synapses occur in this region as a consequence of training. The left and right IMHV regions play different roles in the imprinting process. Exposure to a simple artificial object, a rotating red box, has different neural consequences from those associated with exposure to a complex object, a rotating stuffed jungle fowl, which resembles a conspecific. These differences may be related to the differences in complexity of the two training objects. Another possibility is that two neural systems are implicated in imprinting: a system that underlies a predisposition to approach objects resembling conspecifics and a learning system, of which IMHV is a crucial component, that is engaged by particular objects and that in "natural" circumstances also allows the chick to learn the characteristics of its mother.

Changes in the structure of synapses associated with learning

Two experiments were conducted to investigate the effects of training on a region of the chick brain known to be critically involved in imprinting, the intermediate and medial extent of the hyperstriatum ventrale (IMHV). In the first experiment, three groups of chicks were used: (i) dark-reared (n = 9), (ii) trained for 20 min (n = 17), and (iii) trained for 140 min (n = 7). Chicks were trained by exposing them when they were approximately 21 hr old to a flashing red light. Chicks were killed when they were approximately 30 hr old and blocks of tissue were removed from the right and left IMHV. Stereological techniques were used to measure from electron micrographs the numerical density of dendritic spine and shaft synapses and the length of the postsynaptic density of these synaptic junctions. There was a significant effect of training only in the left IMHV and on only one measure, the overall mean length of the postsynaptic density of spine synapses, SPL. This measure was significantly greater by 17.2% in chicks trained for 140 min than in dark-reared controls. There was no significant effect of training for 20 min. In the second experiment one group of chicks (n = 15) was exposed to a rotating red box for a total of 3 hr. Another group of chicks was dark-reared (n = 15). The chicks were killed when they were approximately 46 hr old. Samples from the hyperstriatum accessorium and IMHV of the right and left sides were analyzed. Training was associated with a significant change, an increase, only of SPL in the left IMHV.(ABSTRACT TRUNCATED AT 250 WORDS)