Acute intrahippocampal infusion of BDNF induces lasting potentiation of synaptic transmission in the rat dentate gyrus.

The effect of acute intrahippocampal infusion of brain-derived neurotrophic factor (BDNF) on synaptic transmission in the dentate gyrus was investigated in urethan-anesthetized rats. Medial perforant path-evoked field potentials were recorded in the dentate hilus and BDNF-containing buffer was infused (4 microl, 25 min) immediately above the dentate molecular layer. BDNF led to a slowly developing increase of the field excitatory postsynaptic potential (fEPSP) slope and population spike amplitude. The potentiation either reached a plateau level at approximately 2 h after BDNF infusion or continued to increase for the duration of experiment; the longest time point recorded was 10 h. Mean increases at 4 h after BDNF infusion were 62.2 and 224% for the fEPSP slope and population spike, respectively. No changes in responses were observed in controls receiving buffer medium only or buffer containing cytochrome C. BDNF-induced potentiation developed in the absence of epileptiform activity in the hippocampal electroencephalogram or changes in recurrent inhibition on granule cells as assessed by paired-pulse inhibition of the population spike. We conclude that exogenous BDNF induces a lasting potentiation of synaptic efficacy in the dentate gyrus of anesthetized adult rats.

Heterosynaptic LTD and depotentiation in the medial perforant path of the dentate gyrus in the freely moving rat.

We examined the characteristics of heterosynaptic long-term depression (LTD) and depotentiation of previously established long-term potentiation (LTP) in the medial and lateral entorhinal afferents to the dentate gyrus in the awake rat. Rats were prepared for chronic recording of dentate gyrus evoked potentials to activation of the medial and lateral perforant paths. This study in awake rats confirms that heterosynaptic LTD can be induced at inactive medial perforant path synapses in conjunction with the induction of LTP produced by high-frequency stimulation of the lateral perforant path. This form of LTD was long lasting and reversible by tetanic stimulation delivered to the depressed pathway. In contrast, tetanic stimulation of the medial perforant path had only a small heterosynaptic effect on the lateral pathway, suggesting that the two input pathways to the dentate gyrus are not symmetrical in their ability to induce heterosynaptic LTD. We also examined the ability of high-frequency stimulation of one pathway to produce depotentiation of the other pathway. We found that when LTP was first induced in the medial perforant path, depotentiation was induced heterosynaptically by tetanization of the lateral pathway. Both newly established LTP (30 min) and LTP induced and saturated by repeated tetanic stimulation over several days can be depotentiated heterosynaptically. Moreover, depotentiation of the medial perforant path synapses was found to be linearly correlated with the magnitude of LTP induced in the lateral perforant path synapses, and subsequent tetanic stimulation of the depotentiated medial perforant path restored LTP to an extent that counterbalanced depotentiation. The saturation and repotentiation experiments provide clear support for the conclusion that the rapid reversal of LTP reflects true depotentiation of the medial input. Again, as with heterosynaptic LTD, tetanization of the medial perforant path had little effect on previously induced LTP in the lateral path. These results provide evidence that medial perforant path synapses can be depressed and depotentiated heterosynaptically. They suggest that in the intact rat synaptic changes in the afferents to the dentate gyrus from the lateral entorhinal cortex exert powerful control over ongoing or recent synaptic plasticity in the medial entorhinal afferents.

Delta opioid receptor activation is required to induce LTP of synaptic transmission in the lateral perforant path in vivo.

The role of opioid receptors in long-term potentiation (LTP) of the medial (MPP) and lateral (LPP) divisions of the perforant path-granule cell projection was investigated in urethane anesthetized rats. A stimulating electrode was positioned in the dorsomedial or ventrolateral aspect of the angular bundle for selective activation of the MPP and LPP, respectively. A push-pull cannula served to focally perfuse artificial cerebrospinal fluid (ACSF) across the perforant path terminal zone, while perforant path evoked potentials were monitored in the dentate hilus. Robust LTP of the excitatory postsynaptic potential (EPSP) initial slope and population spike height was induced by high frequency stimulation (400 Hz, 8 bursts of 8 pulses) applied to the medial or lateral perforant path in rats perfused with standard medium. In the lateral perforant path, a putative proenkephalin system, LTP of the EPSP and population spike was blocked when ACSF containing 100 microM of the opioid receptor antagonist naloxone was present during the tetanus, while perfusion with 0.1 microM naloxone prevented EPSP potentiation but only reduced the magnitude of the population spike increase. Naloxone had no effect on LTP induction in the MPP. Importantly, 0.1 microM ICI 174,864, a selective antagonist of delta opioid receptors, blocked LTP of synaptic transmission in the LPP while leaving the population spike increase intact. The results indicate that LTP of synaptic transmission in the LPP requires activation of delta opioid receptors, while 'non-delta' opioid receptors may be involved in augmenting granule cell output. This opioid receptor-dependent LTP illustrates peptidergic regulation of synaptic plasticity in the hippocampus.

Synaptic plasticity in the hippocampus is modulated by behavioral state.

The possible influence of the sleep-waking cycle on evoked neurotransmission and on the induction of long-term potentiation (LTP) and depression (LTD) was studied in the perforant path-granule cell system. Freely moving rats received a high-frequency stimulus train (8 bursts at 400 Hz) during slow-wave sleep (SWS), rapid eye movement (REM) sleep, and a still-alert (SAL) behavioral state. Trains applied during SAL and REM reliably elicited LTP of the excitatory postsynaptic potential (EPSP) slope, population spike height, and spike onset latency. Granule cell excitability was also enhanced, as indicated by a leftward shift of the EPSP-population spike (E-S) relation. In contrast, tetanization in SWS rarely produced 'classical' LTP and often failed to elicit any lasting change in field potentials. Furthermore, the following types of E-S change occurred almost exclusively after tetanization in SWS: (1) LTP of the EPSP accompanied by depression of the population spike, and (2) E-S potentiation without a change in EPSP. When LTP occurred, however, its magnitude was independent of the animal's behavioral state at the time of the train. In agreement with previous reports, the efficacy of low-frequency neurotransmission varied with behavioral state. A modulation index (MI) was introduced to quantify the difference between field potentials evoked in SAL and SWS. Interestingly, both the occurrence and magnitude of LTP were related to the strength of the MI, as determined in each rat before the train. After trains, the state-dependent modulation of transmission was maintained and was superimposed on LTP and LTD. The results suggest that synaptic plasticity is dynamically modulated during the sleep-wakefulness cycle.

Induction of long-term depression and potentiation by low- and high-frequency stimulation in the dentate area of the anesthetized rat: magnitude, time course and EEG.

We investigated the possible importance of stimulus train frequency for the induction and magnitude of long-term synaptic plasticity in the perforant path-granule cell pathway. Under the same experimental conditions, low- (15 Hz) or high-frequency (400 Hz) stimulation could elicit a profound long-term depression (LTD), or typical long-term potentiation (LTP), of the population spike amplitude, excitatory postsynaptic potential (EPSP) amplitude and spike onset latency. In addition, changes in the relationship between the EPSP and population spike amplitude indicated that granule cell excitability was enhanced during LTP and reduced during LTD. LTD occurred primarily after low-frequency stimulation (5 of 6 cases), and was always accompanied by striking changes in the EEG, most notably a biphasic slow potential. While the EEG changes were confined to the first 5 min after the tetanus, LTD lasted from 1 to 4 h. The nature of the EEG events is still unclear, it is suggested that they may represent a spreading depression-like episode. Finally, we found that LTP evoked by high-frequency stimulation was larger and generally reached peak magnitude faster than when it followed low-frequency stimulation. A possible mechanism and role for hippocampal LTD is proposed.

Effect of 5-HT depletion of the hippocampus on neuronal transmission from perforant path through dentate gyrus.

Stimulation of the perforant pathway (pp) elicits a characteristic evoked action potential (EAP) in the granule cell layer of the dentate gyrus. The EAP was recorded in rats depleted of hippocampal serotonin (5-HT) by prior injection of p-chloroamphetamine (PCA) or 5,7-dihydroxy-tryptamine (5,7-DHT) as well as in untreated animals during two behavioral states, slow-wave sleep (SWS) and still-alert behavior (SAL). As reported previously, in untreated rats the amplitude of the EAP response was significantly greater during SWS than SAL. Stimulation of the median raphe nucleus (MR) prior to stimulating the pp (prestimulation) augmented the EAP response, but only during SWS. In contrast, in animals injected with PCA or 5,7-DHT there was no difference of the amplitude of the EAP during SWS and SAL. However, the augmentation of the EAP during SWS produced by prestimulation of the median raphe was still present. It is concluded that 5-HT innervation of the dentate gyrus may be involved in the behavioral modulation of the EAP response. Modulation of the EAP following prestimulation of the MR appears to be effected by a non-serotonergic input to the dentate gyrus originating in, or coursing through, the median raphe.

Suprachiasmatic lesions disrupt the daily rhythmicity in the sexual behaviour of normal male rats and of male rats treated neonatally with antioestrogen.

Male rats were treated daily with oil or 100 micrograms of the antioestrogen, ethamoxytriphetol (MER-25), for the first 10 days of life and, when adult, lesions were made in the suprachiasmatic nuclei (SCN) of the hypothalamus or control lesions were made above the SCN and the rats were tested for sexual behaviour. Treatment with MER-25 enhanced the daily rhythmicity in both mounting and lordosis behaviour and SCN lesions disrupted these behavioural rhythms and the rhythm in the mounting behaviour of oil-treated rats. Rats treated with MER-25 and with SCN lesions showed high levels of mounting and lordosis behaviour throughout the light : darkness cycle. These results support the hypothesis that sexual differentiation by perinatal androgen stimulation uncouples the central rhythm generator from the neural substrates of sexual behaviour in rats.

Effects of lateral and medial septal lesions on exploratory behavior in the albino rat.

The effects of selective medial and lateral septal lesions on exploratory behavior were studied in the rat. Three types of open-field test and two tests of object exploration were used. In the first experiment, medial but not lateral septal lesions abolished exploration of an open-field accessible from the home-cage. No effects of either of the lesions were found in a standard open-field (Experiment 2). In the third experiment, rats with medial septal lesions were less active and showed a preference for the smaller compartments of the testing apparatus. These results suggest that a change in exploratory behavior following medial septal lesions may be primarily due to their effect on emotionality (increased fear). Two tests of object exploration (Experiments 4 and 5) showed that both types of selective septal lesions enhanced approaches to the novel object placed in the home-cage, but they did not affect object exploration and object preference in other conditions (Experiment 5). The same experiments revealed an increased level of activity (rearing and ambulation) in the rats with lateral septal lesions. It is concluded that the effect of medial septal lesions on exploration is due to increased fear of novel places (but not objects). Lateral septal lesions, on the other hand, increase activity in the rats but there is no evidence that this activity represents enhanced exploration.

A sexually dimorphic rhythm in oestradiol-activated lordosis behaviour in the rat.

Ovariectomized rats exposed to constant plasma levels of oestradiol showed a daily rhythm in lordosis behaviour, with high levels of lordosis occurring during the dark portion of the daily light: darkness cycle and low levels during the light period. Similarly treated male rats failed to show a rhythm in lordosis behaviour. However, neonatal castration permitted the expression of the lordosis rhythm in male rats; conversely, an injection of 1.25 mg testosterone propionate on day 4 of life abolished the rhythm in female rats. Pinealectomy, adrenalectomy or depletion of brain 5-hydroxytryptamine levels did not affect the periodicity in lordosis behaviour but lesions in the suprachiasmatic nuclei of the hypothalamus disrupted the rhythm. It is suggested that the daily rhythm in lordosis behaviour participates in the control of the termination of heat in the female rat and that the perinatal hormone milieu may exert permanent effects on periodic functions.

A daily rhythm in the behavioural sensitivity of the female rat to oestradiol.

Ovariectomized rats were implanted with oestradiol-filled elastomer capsules which were removed at various times after implantation. Sexual receptivity was tested after implantation of a progesterone-filled elastomer capsule 42 h after the onset of treatment with oestradiol. Exposure to oestradiol for about 32 h was required for induction of receptivity. Intact rats with regular 4 day oestrous cycles, and exposed to oestradiol-filled elastomer capsules for 6 h, showed sexual receptivity 24 h after the onset of oestradiol treatment. The behavioural effects of oestradiol in both ovariectomized and intact rats depended on when during the light-darkness (LD) cycle stimulation with oestradiol occurred; maximum effects were seen only if the oestradiol capsules were implanted at 16.00 h (4 h after lights off in the 12 h L : 12 h D cycle). The behavioural effect of progesterone implants, however, did not depend on the phase of the LD cycle. The LD-dependent rhythm in oestradiol sensitivity was eliminated by lesions in the suprachiasmatic nuclei of the hypothalamus.

Some retrohippocampal afferents to the entorhinal cortex. Cells of origin as studied by the HRP method in the rat and mouse.

Small HRP injections were made in the medial (MEC) and lateral entorhinal cortex (LEC) of rat and mouse. Injections in the MEC produced bilateral labeling of cells in layers II and III of the presubiculum as well as in the ipsi- and contralateral parasubiculum. There was no detectable transport of HRP to cells in pre- and parasubiculum following similar injections in the LEC. Injections in the deep layers (IV-VI) of MEC and LEC caused labelling of cells in the ipsilateral subiculum.

Some enzyme histochemical characteristics of the human hippocampus.

The histochemical distribution of acetylcholinesterase (AChE) and alpha-glycerophosphate dehydrogenase (alpha-GPDH) was studied in the area dentata and hippocampus proper of the human brain. Although differences did exist, there were many features in common with the distribution of these two enzymes in the rat and guinea pig. The laminar chemoarchitectonic picture was not as distinct in the human brain as in the rat and guinea pig. Most of the AChE reaction products were confined to the neuropil, with the strongest staining intensity in supra- and infrapyramidal zones. The layer of mossy fibres (stratum lucidum), was characteristically pale. On the other hand, AChE-positive cell bodies were observed in the hilus of the area dentata and a few scattered cells in the hippocampus proper. The AChE reaction products were sparse in the pyramidal cells, conforming similar observations in the rat and guinea pig. Based on our previous description of the AChE-positive cell bodies and fibres in the human septum and a considerable body of experimental material obtained in the rat, it is suggested that most of the AChE in the dentate area and the hippocampus proper is confined to terminals of cholinergic septal efferents in both man and other species. alpha-GPDH was particularly reactive in the cell layers, hilus fasciae dentatae and the layer of mossy fibers. These observations are similar to those described for the rat and guinea pig. They indicate, furthermore, a particular metabolic property common to the archicortex of man and other species.

Activity, avoidance learning and regional 5-hydroxytryptamine following intra-brain stem 5,7-dihydroxytryptamine and electrolytic midbrain raphe lesions in the rat.

Rats underwent one of the following treatments: (1) electrocoagulation of both the dorsal and median midbrain raphe nuclei; (2) 5,7-dihydroxytryptamine creatinine sulfate (5,7-DHT) injection (10 mug, as the salt, in 5 mul vehicle) into the vicinity of each midbrain raphe nucleus; (3) intra-brain stem vehicle (5 mul of 0.2% ascorbic acid in isotonic saline) injections; or, (4) a control operation. Open field activity and one-way avoidance conditioning were examined on postoperative days 16-23. Regional central 5-hydroxytryptamine (5-HT) and catecholamine (CA) concentrations were determined 25-27 days postoperatively. Regional 5-HT levels were greatly reduced following 5,7-DHT administration and electrolytic raphe lesions. The 5,7-DHT rats also showed a reduction in spinal 5-HT content. Central CA concentrations were not affected. Variation in the pattern of regional 5-HT changes after 5,7-DHT treatment was observed but appeared to be related to the adequacy of the dorsal raphe (B7) injection. Only the electrolytic raphe lesion animals, however, showed increased locomotor activity and retarded acquisition and forced-extinction of the one-way avoidance response. In contrast, no significant differences were observed in the open field and avoidance behavior of the 5,7-DHT, vehicle, and control groups. The hyperactivity and impaired one-way avoidance performance observed after electrolytic midbrain raphe lesions are not related simply to reductions in regional forebrain 5-HT and may well be due to damage of non-serotonergic neural systems. Clearly, the behavioral effects of central 5-HT depletion depend on the method employed. The role of 5-HT in regulating activity level and mediating avoidance behavior, furthermore, remains to be determined.

5-Hydroxytryptamine depletion and avoidance acquisition in the rat. Antagonism of the long-term effects of p-chloroamphetamine with a selective inhibitor of 5-hydroxytryptamine uptake.

Twenty four male Sprague-Dawley albino rats in three equal (n = 8) groups were injected on two consecutive days with either p-chloroamphetamine hydrochloride (PCA) (10 mg/kg), H 102/09, an inhibitor of 5-hydroxytryptamine (5-HT) uptake (20 mg/kg) half an hour before PCA, or saline (1 ml/kg). Seven days later the acquisition of a two-way active avoidance response was studied in 8 daily sessions of 20 trials. PCA injections reduced 5-HT concentration of the whole brain 64% at the end of testing (15th day) whereas pretreatment with H 102/09 completely blocked this effect. PCA-treated rats were severely retarded in learning the avoidance response but there was no change in the acquisition of the escape. In contrast, rats pretreated with H 102/09 acquired the avoidance response as rapidly as the saline control group. These results suggest that the impaired learning of the two-way avoidance response may be related to the long-term effects of PCA on the 5-HT neurons in the rat brain. It is concluded that 5-HT neurons may play a role in acquisition of behaviors motivated by aversive events.

Behavioral effects of selective midbrain raphe lesions in the rat.

Lesions were produced in the median (n = 8), dorsal (n = 7) or both (n = 7) midbrain raphe nuclei and their effects on behavior (days 16-54 postoperatively) compared to that of controls (n = 9). In addition, forebrain 5-hydroxytryptamine (5-HT) concentration were determined. Only the median and combined lesion groups showed increased running wheel and open field activity, as well as enhanced reactivity to novel stimuli and environmental change. None of the lesion groups, however, showed changes in home cage activity on postoperative day 21. Although all lesion groups were deficient in the acquisition and retention of one-way avoidance, the deficits were of a greater magnitude in the median and combined lesion groups. The latter two groups, furthermore, were impaired in forced extinction of the one-way avoidance response, but only the combined lesion group evidenced facilitation of two-way avoidance acquistion. Thus, in contrast to the effects of median or combined raphe lesions, lesions in the dorsal raphe nucleus affected few of the behavioral parameters studied. These results suggest that the dorsal raphe nucleus plays a different behavioral role than the median raphe nucleus. The median nucleus appears to be involved in the regulation of activity level, the reaction to novelty and environmental change, and the response to aversive stimuli. Possible mechanisms for the observed behavioral changes are discussed, as well as their apparent similarity to the effects of other mesencephalic and limbic lesions. Lastly, the median, dorsal and combined raphe lesions lowered forebraine 5-HT but 26, 65, and 77%, respectively, versus controls. These reductions differed significantly from each other, and with previously reported data indicate that the dorsal raphe nucleus in the principal origin of forebrain 5-HT. It is suggested, furthermore, that the behavioral effects of midbrain raphe lesions are not due primarily to their associated reduction in forebrain 5-HT.