Effects of early experience on regional incorporation of precursors into RNA and protein in the chick brain.
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One-day-old chicks were exposed for 60 min to either an imprinting stimulus or a dark-box. They were killed at 0, 1, 6 or 12 h after the end of this treatment, and the activities of acetylcholinesterase (AChE) and choline acetyltransferase (ChAc) assayed in the forebrain roof, forebrain base and midbrain. The activity of AChE increased by 11% in the roof of stimulus-exposed birds 1 h after exposure and in base (by 13%) and midbrain (by 8%) 6 h after exposure. At 12 h the only difference was a lowered AChE (14%) activity in midbrain of stimulus-exposed animals. By contrast only one difference was found in ChAc activity between the two types of birds: an elevation of 10% immediately after the end of treatment (0 h) in the midbrain of the stimulus-exposed birds.
The specific activity of RNA polymerase was measured in nuclei from 3 brain regions of 1-day-old chicks which had been exposed for different periods to either an imprinting stimulus or diffuse light and compared to that in birds kept in the dark. Several differences were observed, all of which were short-lived. After 15 min treatment there was a decrease in enzyme activity in the forebrain base of both stimulus- and diffuse light-exposed birds as compared to dark birds, which disappeared by 30 min treatment. After 30 min treatment there was increased activity of the enzyme in the forebrain roof of the stimulus-exposed as compared to both diffuse light-exposed and dark-maintained birds. There were no differences in activity in any region from 45 min up to 120 min of treatment. These changes in RNA polymerase activity were in general accord with the altered uracil incorporation into RNA measured in previous experiments where birds were exposed to 38 or 76 min of treatment, and also with our earlier report on RNA polymerase activity after 30 min treatment.
On the first day after hatching 106 domestic chicks were injected with (14-C)uracil and exposed to a yellow or a red rotating flashing light for 72 min. Shortly before they were killed and 115 min after injection they were given a choice between the two lights. Incorporation of the (14-C)uracil into 4 regions of the brain was measured. Differences between batches and between training conditions were eliminated as sources of variation by standardising the behavioural and biochemical measures. Correlations between the measures were obtained. Chicks that were active from the early stages of training showed lower rates of incorporation in the midbrain, forebrain base and anterior of roof of the forebrain, than less responsive chicks. When the general lowering effect of behavioural activity on incorporation was eliminated as a source of variations, a striking positive correlation emerged between the preference for the familiar object and incorporation in the anterior of forebrain roof. Since preference for the familiar is a direct measure of imprinting and since no other behavioural measure is positively correlated with biochemical changes in anterior roof, it is concluded that these biochemical changes were closely linked with the learning process.
On the first day after hatching domestic chicks were exposed to an imprinting stimulus, a horizontal yellow slit of light moving upwards in a window and presented at a rate of 4 slits/sec. Chicks were exposed for either 45 min (undertrained) or 180 min (overtrained) on the first day of hatching (60 or 240 min in the case of 1 pair). On the second day all birds were exposed for a further 63 min. Twenty birds were matched in pairs and each chick received 1.1 muCi [14C]uracil/g body weight injected into the heart region before exposure on day 2. At 150 min after the injection the chicks were decapitated and serial coronal sections of their brains cut; alternate pairs of sections were prepared for autoradiography. The optical density for a number of major anatomical regions was measured. The measurements for each region were averaged over congruent to 0.6 mm 'slabs' of brain and expressed as a percentage of the mean of all measurements for that brain. Standardized mean optical density was significantly greater in undertrained chicks than in overtrained chicks in a part of the medial region of hyperstriatum ventrale (MHV) which extended across two adjacent slabs. The slabs were slightly posterior to the mid-point between anterior and posterior poles of the brain. In the next two, adjacent, anterior slabs the variances for MHV were significantly greater in undertrained chicks than in overtrained chicks. There were no other significant differences between brain regions. Taken together with previous studies these results suggest that the intermediate and medial part of hyperstriatum ventrale is intimately linked with the imprinting process.
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The effects of placing bilateral lesions in that part of the chick brain (IMHV) which was previously been implicated in imprinting, was studied in young domestic chicks. Twenty-four dark-reared chicks were matched in pairs on the basis of their approach activity during a 30 min period of exposure to one of two visual imprinting stimuli. Both members of the chick pair were then anaesthetized and bilateral lesions were made by radio-frequency coagulation in the IMHV of one chick; the other chick served as a sham-operated control. On the following day each chick was exposed for 2.5 h to the imprinting stimulus to which it had previously been exposed. After training, the preferences of all chicks were measured by comparing their approach to the training stimulus with that to the second stimulus. Sham-operated chicks showed a strong preference for the training stimulus; lesioned chicks showed none. Subsequently the latency of each chick to approach and accurately peck a shiny rod was measured. The two groups of chicks did not differ significantly in this test of visuomotor coordination. The area of tissue damaged by the lesion was reconstructed: IMHV was severely damaged with relatively little damage to other areas of the brain.
Young chicks were placed in running wheels and exposed for a total of 3 h to an imprinting stimulus: a rotating flashing red light. Revolutions made towards the stimulus minus revolutions made away from it were totalled and are referred to as approach counts. After exposure, chicks were anaesthetized and two vertical electrode penetrations were made, one through the visual projection area, hyperstriatum accessorium (HA), and one through the intermediate and medial part of hyperstriatum ventrale (IMHV), a region which has been implicated in the imprinting process. 'Spontaneous' impulse activity was recorded from neurones in these areas. In some of the chicks, after penetrating through HA, discharges evoked by diffuse retinal illumination were monitored and the zona hyperstriatum intercalatus/hyperstriatum dorsale (HI/HD) below HA was classified as being a visually responsive lamina (VRL) if responses were obtained to greater than or equal to 20 consecutive light flashes. In HA there was a positive correlation between spontaneous activity and approach counts in those chicks with a VRL; there was no significant correlation in those chicks without a VRL. In contrast, for IMHV the correlation between spontaneous activity and approach counts was negative and appeared to be independent of the presence or absence of a VRL.
The maturation of the granule cells of the ferret olfactory bulb around the time of odour imprinting has been examined. Rapid Golgi impregnation studies revealed a temporal overshoot in the development of the spines on the external and internal dendrites of the granule cells. In contrast, the number of somatic spines decreased continuously. Electron microscopical examinations of the synaptic contacts in the external plexiform layer revealed that the time course of synapse and reciprocal synapse formation was similar to that of the formation of the spines on the external dendrites. The results were taken as evidence that both the Golgi and the electron microscopical investigations described the same developmental process of postnatal synaptic rearrangement.
The distribution and morphology of neurons containing the Ca-binding proteins parvalbumin (PV), calbindin-D28k (CaBP) and calretinin (CaR) are described in a rostral forebrain region (MNH) of the chick, known to be involved in auditory filial imprinting. PV immunoreactivity is chiefly a marker for numerous large to medium-sized neurons in the neostriatal part of MNH. They show patchy staining of their dendrites, but PV-positive spines are not visible. CaBP is represented in a different neuron population with on the average slightly smaller-sized somata, which carry long, spiny, CaBP-positive dendrites. In contrast to PV and CaBP, CaR immunoreactivity is a marker chiefly for neuropil in MNH but only for few stained neurons. They may be spiny and show the largest size variations. The density of CaR-immunoreactive neuropil is highest in the hyperstriatal part of MNH. Double immunostaining for PV and CaBP reveals that these proteins are expressed mostly in different neuron populations, with only few neurons containing both proteins. These neuron populations appear to form an interconnected network within MNH. A possible relationship between the expression of either Ca-binding protein and the presence of the inhibitory transmitter GABA is also examined. The GABA-antibody labels scattered, very small to medium-sized neurons and dense punctate neuropil. The comparison of the area histograms of somata reveals an overlap with all 3 Ca-binding protein containing cell populations, except for a large proportion of small GABA-positive neurons. The characteristics of immunostained neuron populations are compared to the previously described 3 Golgi-types of neurons in MNH, and possibilities of a functional implication of the proteins in MNH plasticity are examined.
Bilateral ablation of the telencephalon on the 1st posthatch day prevented learning from imprinting to colored stimuli in Japanese quail chicks C. coturnix japonica, but it spared their artificially selected approach preferences for particular colors and patterns. Post-imprinting decerebration on the 3rd posthatch day erased the imprinted memory, but did not revert the chicks' approach choices to the colors of their genetically distinct initial preferences. The genetically determined and the acquired preferences were both lost in the latter birds, regardless whether they were imprinted to the initially preferred or the initially nonpreferred stimuli before decerebration. Discussion focuses on implications of the data for modeling specific interactions between and the interrelated neural representation of genetic and environmental influences in early behavioral development.
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.
Vasoactive intestinal peptide (VIP) is a neuromodulator and secretagogue for neuronal survival factors. Moreover, in the suprachiasmatic nucleus (SCN) of mature animals, where the circadian clock resides, day-night cycles are paralleled by rhythmical alteration in the levels of the mRNA encoding VIP. We now show, for the first time, that diurnal oscillation in VIP mRNA occurred in animals reared in the dark and also preceded eye opening. Thus, by Northern blot hybridization elevated levels of VIP mRNA were measured at night time as compared to day time. Furthermore, diurnal oscillation in VIP mRNA were more pronounced in rats reared in the dark. These observations suggested an endogenous as well as exogenous (environmental) control of VIP gene expression in the SCN and implicated VIP as an integral part of the circadian clock.
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 anaesthetized 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.