Tonic Immobility In Guinea Pigs Research Articles

Similar effect of CRF1 and CRF2 receptor in the basolateral or central nuclei of the amygdala on tonic immobility behavior

Studies have used paradigms based on animal models to understand human emotional behavior because they appear to be correlated with fear- and anxiety-related defensive patterns in non-human mammals. In this context, tonic immobility (TI) behavior is an innate response associated with extreme threat situations, such as predator attack. Some reports have demonstrated the involvement of corticotropin-releasing factor (CRF) in regulation of the endocrine system, defensive behaviors and behavioral responses to stress. Particularly, a previous study showed that the activation of CRF receptors in the basolateral (BLA) or central (CeA) nuclei of the amygdala increased TI responses, whereas treatment with a non-selective CRF antagonist, alpha-helical-CRF9-41, decreased this innate fear response. However, while CRF1 receptors have pronounced effects in stress-induced anxiety, CRF2 receptors appear be involved in the expression of both stress-induced anxiety and spontaneous anxiety behavior. In this study, we investigated the effects of specific CRF receptors, CRF1 and CRF2, in the BLA and CeA on the duration of TI in guinea pigs. The results show that blockade of CRF1 and CRF2 receptors in the BLA and CeA produces a decrease in fear and/or anxiety, as suggested by a decrease in TI duration in the guinea pigs. Additionally, the specific antagonists for CRF1 and CRF2 receptors were able to prevent the increase in TI duration induced by CRF administration at the same sites. These results suggest that the modulation of fear and anxiety by the CRF system in the BLA and CeA occurs through concomitant effects on CRF1 and CRF2 receptors.

Effects of human contact and intra-specific social learning on tonic immobility in guinea pigs, Cavia porcellus

Social learning is the capacity of animals to acquire adaptive information from others. In the case of fear responses, animals can learn fearful or non-fearful responses by observing the behavior of conspecifics. Tonic immobility (TI) is an anti-predatory behavior elicited during intense fear situations. Studies have revealed that regular contact with humans can reduce TI responses in animals. In our study, we evaluated the effect of human contact on the TI responses in guinea pigs. We also evaluated the effect of cohabitation (non-fearful animals with fearful animals) on their TI responses. To achieve this, we measured the TI responses induced by postural inversion and restraint in guinea pigs as a result of different treatments. In our first experiment, we determined the effect of human contact on TI responses by establishing 3 treatment groups: no contact, handled, and habituated. In our second experiment, we addressed the effect of social learning on TI response by testing TI response in habituated, and unhabituated animals that had cohabitated for 10days. In the first experiment, 10days of either handling or habituation did not prevent TI in guinea pigs, but habituation did increase latency [F(2,119)=14.19; p<0.0001] and handling or habituation decrease duration [F(2,119)=15.01; p<0.0001] of the TI behavior in the guinea pigs. In the second experiment, the cohabitation of unhabituated and habituated animals reduced TI duration [F(2,93)=5.058; p<0.008]. These data suggest that both forms of human interaction can reduce experimenter fear in guinea pigs. It therefore seems that unhabituated guinea pigs learn not to fear the experimenter by cohabitating with habituated guinea pigs.

Opioidergic, GABAergic and serotonergic neurotransmission in the dorsal raphe nucleus modulates tonic immobility in guinea pigs

Tonic immobility (TI) is an innate defensive behavior that can be elicited by physical restriction and postural inversion and is characterized by a profound and temporary state of akinesis. Our previous studies demonstrated that the stimulation of serotonin receptors in the dorsal raphe nucleus (DRN) appears to be biphasic during TI responses in guinea pigs (Cavia porcellus). Serotonin released by the DRN modulates behavioral responses and its release can occur through the action of different neurotransmitter systems, including the opioidergic and GABAergic systems. This study examines the role of opioidergic, GABAergic and serotonergic signaling in the DRN in TI defensive behavioral responses in guinea pigs. Microinjection of morphine (1.1nmol) or bicuculline (0.5nmol) into the DRN increased the duration of TI. The effect of morphine (1.1nmol) was antagonized by pretreatment with naloxone (0.7nmol), suggesting that the activation of μ opioid receptors in the DRN facilitates the TI response. By contrast, microinjection of muscimol (0.5nmol) into the DRN decreased the duration of TI. However, a dose of muscimol (0.26nmol) that alone did not affect TI, was sufficient to inhibit the effect of morphine (1.1nmol) on TI, indicating that GABAergic and enkephalinergic neurons interact in the DRN. Microinjection of alpha-methyl-5-HT (1.6nmol), a 5-HT2 agonist, into the DRN also increased TI. This effect was inhibited by the prior administration of naloxone (0.7nmol). Microinjection of 8-OH-DPAT (1.3nmol) also blocked the increase of TI promoted by morphine (1.1nmol). Our results indicate that the opioidergic, GABAergic and serotonergic systems in the DRN are important for modulation of defensive behavioral responses of TI. Therefore, we suggest that opioid inhibition of GABAergic neurons results in disinhibition of serotonergic neurons and this is the mechanism by which opioids could enhance TI. Conversely, a decrease in TI could occur through the activation of GABAergic interneurons.

Role of homocysteic acid in the guinea pig (Cavia porcellus) anterior cingulate cortex in tonic immobility and the influence of NMDA receptors on the dorsal PAG

Tonic immobility (TI) is an innate defensive behaviour elicited by physical restriction and postural inversion, and is characterised by a profound and temporary state of akinesis. Our previous studies demonstrated that glutamatergic stimulation of the dorsomedial/dorsolateral portion of periaqueductal gray matter (dPAG) decreases the duration of TI in guinea pigs (Cavia porcellus). Furthermore, evidence suggests that the anterior cingulate cortex (ACC) constitutes an important source of glutamate for the dPAG. Hence, in the current study, we investigated the effects of microinjection of the excitatory amino acid (EAA) agonist dl-homocysteic acid (DLH) and the N-methyl-d-aspartate (NMDA) receptor antagonist MK-801 into the ACC on the duration of TI in guinea pigs. We also assessed the effect of the NMDA receptor antagonist (MK-801) into the dorsal periaqueductal gray matter (dPAG) prior to DLH microinjection into the ACC on the TI duration in the guinea pig. Our results demonstrated that DLH microinjections into the ACC decreased the duration of TI. This effect was blocked by previous MK-801 microinjections into the ACC or into the dPAG. The MK-801 microinjections alone did not influence TI duration. These results provide the new insight that EAAs in the ACC can decrease the duration of TI. The mechanism seems to be dependent on the NMDA receptors present in the ACC and in the dPAG.

Modulation of tonic immobility in guinea pig PAG by homocysteic acid, a glutamate agonist

Tonic immobility (TI) is an innate defensive behavior elicited by physical restriction and postural inversion, and is characterized by a profound and temporary state of motor inhibition. The participation of the periaqueductal gray matter (PAG) in TI modulation has previously been described. In addition, the excitatory amino acids (EAA) are important mediators involved in the adjustment of several defensive responses produced by PAG. In the present study, we investigated the effect of microinjection of the EAA agonist dl-homocysteic acid (DLH) and the N-methyl- d-aspartate (NMDA) receptor antagonist (MK-801) into the ventrolateral and dorsal PAG over the duration of TI in guinea pigs. Microinjection of 15 nmol/0.2 µl of DLH into the ventrolateral PAG (vlPAG) and 30 nmol/0.2 µl of DLH into the dorsal PAG (dPAG) promoted an increase and decrease in TI duration, respectively. These responses were blocked by prior microinjection of the NMDA receptor antagonist, MK-801 (3.6 nmol/0.2 µl) at the same site. Microinjection of MK-801 alone into the vlPAG and dPAG did not alter the duration of TI episodes. These results suggest that NMDA receptors are involved in the modulation of TI in both the vlPAG and dPAG. In addition, PAG excitatory amino acids modulate the TI response via columnar organization of the PAG. In this manner, the vlPAG facilitates TI modulation whereas dPAG has an inhibitory role in TI.

Role of opioidergic and GABAergic neurotransmission of the nucleus raphe magnus in the modulation of tonic immobility in guinea pigs

Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. Previous results from our laboratory have demonstrated that nucleus raphe magnus (NRM) is also a structure involved in the modulation of TI behavior, as chemical stimulation through carbachol decreases the duration of TI in guinea pigs. In view of the fact that GABAergic and opioidergic circuits participate in the regulation of neuronal activity in the NRM and since these neurotransmitters are also involved in the modulation of TI, the objective of the present study was to evaluate the role of these circuits of the NRM in the modulation of the behavioral TI response. Microinjection of morphine (4.4nmol/0.2μl) or bicuculline (0.4nmol/0.2μl) into the NRM increased the duration of TI episodes while muscimol (0.5nmol/0.2μl) decreased it. The effect of morphine injection into the NRM was blocked by previous microinjection of naloxone (2.7nmol/0.2μl). Muscimol at 0.25nmol did not produce any change in TI duration; however, it blocked the increased response induced by morphine. Our results indicate a facilitatory role of opioidergic neurotransmission in the modulation of the TI response within the NRM, whereas GABAergic activity plays an inhibitory role. In addition, in the present study the modulation of TI in the NRM possibly occurred via an interaction between opioidergic and GABAergic systems, where the opioidergic effect might be due to inhibition of tonically active GABAergic interneurons.

Cholinergic modulation of tonic immobility and nociception in the NRM of guinea pig

Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. It is known that endogenous antinociceptive systems are activated during the emission of defensive behaviors including TI. The nucleus raphe magnus (NRM) is related to the modulation of nociceptive and behavioral responses. In the present study, we investigated the role of the cholinergic system of the NRM in the modulation of TI and nociception in guinea pigs. Microinjection of the cholinergic agonist carbachol (0.5 μg/0.2 μl) into the NRM promoted a reduction in the duration of TI episodes and nociception, the latter measured by the vocalization test in guinea pigs. The effect of microinjection of carbachol on TI reduction and antinociception was blocked by the previous microinjection of the cholinergic antagonist atropine (0.5 μg/0.2 μl and 1 μg/0.2 μl, respectively), demonstrating the participation of muscarinic receptors in the modulation of these responses. Microinjection of atropine per se did not interfere with the duration of TI episodes. In summary, the present results demonstrate that cholinergic stimulation of the NRM promoted analgesia and a reduction in the duration of TI in guinea pigs. These data indicate that the NRM possibly contributes to the modulation of defensive and nociceptive behavioral responses, probably by modulating the activity of neurons in the ventral and dorsal horn of the spinal cord, respectively.

Cholinergic-opioidergic interaction in the central amygdala induces antinociception in the guinea pig.

Several studies have demonstrated the involvement of the central nucleus of the amygdala (CEA) in the modulation of defensive behavior and in antinociceptive regulation. In a previous study, we demonstrated the existence of a cholinergic-opioidergic interaction in the CEA, modulating the defensive response of tonic immobility in guinea pigs. In the present study, we investigated a similar interaction in the CEA, but now involved in the regulation of the nociceptive response. Microinjection of carbachol (2.7 nmol) and morphine (2.2 nmol) into the CEA promoted antinociception up to 45 min after microinjection in guinea pigs as determined by a decrease in the vocalization index in the vocalization test. This test consists of the application of a peripheral noxious stimulus (electric shock into the subcutaneous region of the thigh) that provokes the emission of a vocalization response by the animal. Furthermore, the present results demonstrated that the antinociceptive effect of carbachol (2.7 nmol; N = 10) was blocked by previous administration of atropine (0.7 nmol; N = 7) or naloxone (1.3 nmol; N = 7) into the same site. In addition, the decrease in the vocalization index induced by the microinjection of morphine (2.2 nmol; N = 9) into the CEA was prevented by pretreatment with naloxone (1.3 nmol; N = 11). All sites of injection were confirmed by histology. These results indicate the involvement of the cholinergic and opioidergic systems of the CEA in the modulation of antinociception in guinea pigs. In addition, the present study suggests that cholinergic transmission may activate the release of endorphins/enkephalins from interneurons of the CEA, resulting in antinociception.

Serotonin 5-HT 2 and 5-HT 1A receptors in the periaqueductal gray matter differentially modulate tonic immobility in guinea pig

Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. We have previously reported that cholinergic stimulation of the dorsal periaqueductal gray matter (PAG) decreases the duration of TI episodes, while stimulation of the ventrolateral region increases it. The ventrolateral PAG modulates this behavior via a similar neural circuit proposed to be involved in the antinociceptive system. Some studies have indicated that alterations in the levels of cerebral 5-hydroxytryptamine (5-HT) mediate or modulate the analgesic effect of PAG stimulation. Thus, in this study we investigated the possibility that the serotoninergic system is involved in the modulation of TI by this neural substrate. Our results showed that the effect of serotonin into the ventrolateral and dorsal PAG seems to be biphasic and dose dependent. The microinjection of low doses (0.1 μg) of 5-HT into the PAG increased the duration of TI, while high doses (1, 3 and 6 μg) decreased this behavior. Our results also showed that microinjection of a 5-HT 1A agonist (0.003, 0.01 and 0.1 μg of 8-hydroxy-dipropylaminotretalin (8-OH-DPAT)) into the PAG increased the duration of TI episodes. However, the microinjection of 5-HT 2 agonist (0.01 and 0.1 μg of α-methyl-5-HT) into the PAG decreased the duration of TI and this effect could be reversed by pretreatment with an ineffective dose (0.01 μg) of ketanserin. In contrast, ketanserin (0.03 and 0.16 μg) increased this behavior in a dose-dependent manner. These results suggest that the PAG 5-HT 1A and 5-HT 2 receptors have different roles in the modulation of TI in guinea pigs, since the 5-HT 1A and 5-HT 2 agonists, respectively, increased and decreased the duration of TI.

The cholinergic stimulation of the central amygdala modifying the tonic immobility response and antinociception in guinea pigs depends on the ventrolateral periaqueductal gray

Tonic immobility (TI), also known as death feigning or animal hypnosis, is a reversible state of motor inhibition that is triggered by postural inversion and/or movement restraining maneuvers but also by repetitive stimulation and pressure on body parts. Our previous studies demonstrated that cholinergic stimulation of the central amygdala (CEA) decreases the duration of TI in guinea pigs. Some reports have demonstrated that electrical or chemical stimulation of the CEA promotes antinociception. Evidence suggests that the CEA performs part of its functions by means of a connection with the ventrolateral periaqueductal gray (vlPAG). In the current study, we investigated the participation of a possible functional and anatomical CEA–vlPAG connection in guinea pigs in the regulation of the TI response and antinociception. Our results showed that the functional CEA–vlPAG connection is essential for the participation of the CEA in the modulation of TI and of antinociception. The reversible exclusion of the vlPAG by means of microinjection of 2% lidocaine blocked the inhibitory effect on TI duration and the antinociceptive effect, as determined by a decrease of the vocalization index (VI) obtained with the administration of carbachol (2.7 nmol/0.2 μl) into the CEA. On the other hand, the exclusion of the CEA by lidocaine did not block the antinociception or the increase in TI induced by microinjection of CCh into the vlPAG. Finally, microinjection of the retrograde neurotracer Fast Blue into the CEA or into the vlPAG demonstrated the existence of a reciprocal anatomical connection between the CEA and vlPAG.

Central nucleus of the amygdala and the control of tonic immobility in guinea pigs

Tonic immobility (TI) is a temporary state of profound motor inhibition induced by situations that supposedly generate intense fear, with the objective to protect the animal from attacks by predators. A previous study by our group demonstrated that cholinergic stimulation of the central, basolateral, and lateral posterior nuclei of the amygdala decreases the duration of TI in guinea pigs. In the current study, we attempted to investigate the effects of cholinergic, opioidergic, and γ-aminobutyric acid (GABA)ergic stimulation of the central amygdala (CEA) on TI modulation. We observed that both cholinergic (carbachol, 2.7 nmol/0.2 μl) and opioidergic (morphine, 2.2 and 4.4 nmol/0.2 μl) stimulation of the CEA decreased the duration of TI and that these effects could be reversed by pretreatment with naloxone (1.3 and 2.7 nmol/0.2 μl). Our results also showed that microinjection of the GABAergic agonist muscimol (0.26 nmol/0.2 μl) reduced the duration of TI episodes, whereas microinjection of the GABAergic antagonist bicuculline (BIC, 1 nmol/0.2 μl) increased it. Thus, the present experiments demonstrated that cholinergic, opioidergic, and GABAergic systems of the CEA have an inhibitory action on the duration of TI in guinea pigs. Furthermore, the current study suggests an interaction of cholinergic and opioidergic mechanisms. In addition, the GABAergic circuit of the CEA has a tonic inhibitory influence on the duration of TI and is mediated by GABA A receptors.

Endogenous opiate analgesia induced by tonic immobility in guinea pigs.

A function of the endogenous analgesic system is to prevent recuperative behaviors generated by tissue damage, thus preventing the emission of species-specific defensive behaviors. Activation of intrinsic nociception is fundamental for the maintenance of the behavioral strategy adopted. Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. We studied the effect of TI behavior on nociception produced by the formalin and hot-plate tests in guinea pigs. The induction of TI produced a significant decrease in the number of flinches (18 +/- 6 and 2 +/- 1 in phases 1 and 2) and lickings (6 +/- 2 and 1 +/- 1 in phases 1 and 2) in the formalin test when compared with control (75 +/- 13 and 22 +/- 6 flinches in phases 1 and 2; 28 +/- 7 and 17 +/- 7 lickings in phases 1 and 2). In the hot-plate test our results also showed antinociceptive effects of TI, with an increase in the index of analgesia 30 and 45 min after the induction of TI (0.67 +/- 0.1 and 0.53 +/- 0.13, respectively) when compared with control (-0.10 +/- 0.08 at 30 min and -0.09 +/- 0.09 at 45 min). These effects were reversed by pretreatment with naloxone (1 mg/kg, ip), suggesting that the hypoalgesia observed after induction of TI behavior, as evaluated by the algesimetric formalin and hot-plate tests, is due to activation of endogenous analgesic mechanisms involving opioid synapses.

Role of the Amygdaloid Nuclei in the Modulation of Tonic Immobility in Guinea Pigs

LEITE–PANISSI, C. R. A., C. R. MONASSI AND L. MENESCAL-DE-OLIVEIRA. Role of the amygdaloid nuclei in the modulation of tonic immobility in guinea pigs. PHYSIOL BEHAV 67(5) 717–724, 1999.—Tonic immobility (TI) is thought to represent the terminal reaction in the chain of antipredatory responses involved in maintaining survival. TI is an inhibitory behavioral response in which the animal presents a significant decrease in body activity and responsiveness to the environment induced by some form of physical restraint. This response is induced in the laboratory by inversion of the animal and brief postural contention of its movements. In nature, the TI response may be triggered by some threatening or predatory stimulus, indicating the physical contact between response occurs when there is physical contact between prey and predator. In this case, the physical inactivity of the prey may prevent the continuation of the attack. The neural substrate of this response is not well known, and the objective of the present study was to investigate the effect of cholinergic stimulation of amygdala regions on TI modulation in guinea pigs. Microinjection of carbachol (0.5 μg/0.2 μL) into the central (CEA), basolateral (BLA), and lateral posterior (LPA) nuclei of the amygdala promoted a reduction in the duration of TI episodes. Pretreatment with atropine (0.5 μg/0.2 μL) showed that the action of carbachol is mediated by muscarinic receptors.

The lateral hypothalamus in the modulation of tonic immobility in guinea pigs

The lateral hypothalamus has been reported to be involved in the organization of aggression and predatory attack but not in behavioral inhibition responses such as tonic immobility (TI). TI may be defined as an inborn behavioral inhibition characterized by profound physical inactivity and relative lack of responsiveness to the environment, triggered by an intense sensation of fear generated during prey-predator confrontation. Our study indicates that cholinergic stimulation of anterior regions of the lateral hypothalamus of guinea pigs potentiates the duration of TI episodes, while stimulation of medial and posterior regions of this structure promotes a decrease in TI duration, suggesting that the lateral hypothalamus modulates the duration of TI episodes in a differentiated manner.

Participation of the Medial and Anterior Hypothalamus in the Modulation of Tonic Immobility in Guinea Pigs

Tonic immobility (TI) is an inhibitory behavioral response during which the animal presents profound physical inactivity and a relative lack of response to the environment. This response is induced in the laboratory by postural inversion of the animal and brief postural contention of its movements. In nature, the response occurs when there is physical contact between prey and predator. In this case, the physical inactivity of the prey may prevent the continuation of the attack. The neural substrate of this response is not well known and the objective of the present study was to investigate the effect of cholinergic stimulation of hypothalamic regions on TI modulation in guinea pigs ( Cavia porcellus). Microinjection of carbachol (1.0 μg/0.2 μL) into the anterior hypothalamus promoted an increase in the duration of TI episodes. Microinjection of carbachol into the ventro- and dorsomedial hypothalamus, however, promoted a reduced duration of TI episodes. Pretreatment with atropine (0.5 μg/0.2 μL) showed that the action of carbachol is mediated by muscarinic receptors in the anterior and ventromedial hypothalamus but not in the dorsomedial hypothalamus. The results suggest that the hypothalamic regions may play different roles in the organization of defensive behavioral responses such as TI.

Effects of conditioned aversive stimuli presented during tonic immobility in guinea pigs.

Effects of conditioned aversive stimuli presented during tonic immobility in guinea pigs.

The duration of tonic immobility in guinea pigs.

The duration of tonic immobility in guinea pigs.