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PubMed1992

Intrathecal injection of acetylsalicylic acid, salicylic acid and indomethacin depresses C fibre-evoked activity in the rat thalamus and spinal cord.

It was aimed to assess if intrathecal (i.t.) injections of acetylsalicylic acid and salicylic acid depress C fibre-evoked activity in the sensory part of the nociceptive system. In rats under urethane anaesthesia, activity was elicited in single neurones in the dorsomedial part of the ventral nucleus (VDM) of the thalamus and in ascending axons of the spinal cord by supramaximal electrical stimulation of the sural nerve. Acetylsalicylic acid and salicylic acid injected i.t. significantly reduced the activity evoked in thalamic neurones. The maximum depression amounted to about 50% of the activity evoked in the controls and was produced by acetylsalicylic acid at a dose of 50 micrograms (0.28 mumol)/rat and by salicylic acid at a dose of 37.5 micrograms (0.27 mumol)/rat. Indomethacin injected i.t. also reduced C fibre-evoked activity in the thalamus in a dose-dependent fashion, 100 micrograms producing a 50% depression. Salicylic acid (37.5 micrograms/rat, i.e.) depressed C fibre-evoked activity in ascending axons but had no effect on A beta fibre-evoked activity. It is concluded that i.t. injection of acetylsalicylic acid selectively inhibits nociceptive impulse transmission in the spinal cord by an action of the salicylic acid moiety. It is possible that prostaglandins are involved in the central action of salicylic acid.

Afferent Pathways↗
PubMed1986

A qualitative and quantitative analysis of the distributions of cells in the spinal cord and spinomedullary junction projecting to the thalamus of the rat.

Horseradish peroxidase retrograde transport has been used to locate and make a quantitative study of neurons of the rat spinal cord which project to the thalamus. Across the grey, labelled cells were found in the head and neck of the dorsal horn (in three locations, but not the substantia gelatinosa); in the region of the internal basilar nucleus of Ramón y Cajal; in the intermediate grey zone; in the lateral cervical nucleus and funiculus; and in the ventral horn. The latter contains the largest population of neurons projecting ipsilaterally. The most significant finding concerns the craniocaudal distribution of the cells. More than 50% of the thalamically projecting neurons are confined to the upper four cervical segments, where every population is represented. Some populations are continuous with thalamically projecting populations in the lower medulla. The cervical enlargement contains less than 5% of spinothalamic cells. The lumbar enlargement contains 33%, most of which are in the region of the internal basilar nucleus. This population is also marked in the upper cervical segments. Thus, in the rat, the origin of the spinothalamic tract is distributed along the cord in a very uneven manner. This may indicate that sensory information delivered to any one segment of the spinal cord by a primary afferent is not always relayed direct to the thalamus by a local second order neuron; that different parts of the body are not represented to the same extent in the spinothalamic system; or that these two factors combine to produce the observed distribution.

Afferent Pathways↗
PubMed1991

Membrane and action potential responses evoked by excitatory amino acids acting at N-methyl-D-aspartate receptors and non-N-methyl-D-aspartate receptors in the rat thalamus in vivo.

The membrane potential responses and firing patterns of rat thalamic neurons evoked by iontophoretically applied excitatory amino acids were recorded in vivo. All excitatory amino acids, including N-methyl-D,L-aspartate, evoked a membrane depolarization and a repetitive, regular pattern of action potential firing in the thalamus. Both non-nociceptive and nociceptive thalamic neurons responded to all agonists tested. Iontophoretic application of magnesium ions selectively antagonized responses to N-methyl-D,L-aspartate but did not convert the repetitive firing pattern into a burst firing pattern. In contrast, in the hippocampus, N-methyl-D,L-aspartate evoked a burst pattern of action potential firing associated with rhythmic depolarizing membrane potential shifts, similar to those seen by other workers in the hippocampus and in other brain regions. These findings are discussed in relation to the possibility that the regular firing pattern of spikes evoked by excitatory amino acids in the thalamus is primarily determined by the intrinsic membrane properties of thalamic neurons.

Action Potentials↗
PubMed1993

gamma-Hydroxybutyric acid induced spike and wave discharges in rats: relation to high-affinity [3H]gamma-hydroxybutyric acid binding sites in the thalamus and cortex.

gamma-Hydroxybutyric acid is a naturally occurring compound which induces bilaterally synchronous spike and wave discharges in rats. The gamma-hydroxybutyric acid model of absence seizures simulates clinical absence seizures behaviorally as well as electrographically. The present study was undertaken in order to establish the role of the high-affinity gamma-hydroxybutyric acid binding sites in the generation of gamma-hydroxybutyric acid-induced spike and wave discharges. Spike and wave discharges induced by gamma-hydroxybutyric acid were recorded with the aid of bipolar depth electrodes implanted in discrete regions of thalamus, cortex and hippocampus. In the present study we found that ventroposterolateral, ventroposteromedial, medial and the reticular nuclei of the thalamus discharged synchronously with the cortical generation of spike and wave discharges. In the cortex, the superficial layers (I-IV) of frontoparietal cortex generated spike and wave discharges, whereas no spike and wave discharges were recorded from deeper layers (V-VI) of frontoparietal cortex. At the onset of spike and wave discharges induced by gamma-hydroxybutyric acid, a rapid but reversible upregulation of gamma-hydroxybutyric acid binding sites was observed. This increased [3H]gamma-hydroxybutyric acid binding was characterized by an increase in the number of gamma-hydroxybutyric acid sites with no significant change in their affinity for gamma-hydroxybutyric acid. Moreover, the change in [3H]gamma-hydroxybutyric acid binding was observed only in those thalamic structures and cortical layers which were found to be involved in the generation of spike and wave discharges induced by gamma-hydroxybutyric acid. The CA3 field or dorsal hippocampus possesses the highest density of [3H]gamma-hydroxybutyric acid binding sites of all brain regions. However, no significant change in [3H]gamma-hydroxybutyric acid binding was observed in this region nor was the CA3 field involved in the generation of spike and wave discharges during gamma-hydroxybutyric acid-induced absence-like seizures. These findings confirm that gamma-hydroxybutyric acid-induced absence-like seizures originate from thalamocortical pathways and that the onset of gamma-hydroxybutyric acid-induced spike and wave discharges is directly related to the regulation of gamma-hydroxybutyric acid binding sites in those regions which constitute the involved thalamocortical loop.

Animals↗
PubMed1995

Distinct presynaptic metabotropic receptors for L-AP4 and CCG1 on GABAergic terminals: pharmacological evidence using novel alpha-methyl derivative mGluR antagonists, MAP4 and MCCG, in the rat thalamus in vivo.

A variety of metabotropic excitatory amino acid receptors are present in the thalamus. We have investigated the possibility that some of these receptors may have presynaptic effects on GABAergic inhibitory transmission in the thalamus. Inhibitory responses in ventrobasal thalamic neurons of urethane-anaesthetized rats were evoked by either air-jet stimuli to the vibrissae or by electrical stimulation of the somatosensory cortex. Both intracellular and extracellular recording methods were used to reveal inhibitory responses, either as inhibitory postsynaptic potentials or inhibition of excitatory responses in a condition-test paradigm. The metabotropic glutamate receptor agonists (S)-2-amino-4-phosphonobutyrate (L-AP4) and (2S,3S,4S)-alpha-(carboxycyclopropyl)-glycine (CCG1), applied in the vicinity of the recording site by iontophoresis, were found to reduce the amplitudes of inhibitory postsynaptic potentials (to 76% and 63% of control amplitudes, respectively) and inhibitions revealed by the condition-test paradigm (to 33% and 28% of control inhibitions, respectively). As the inhibitory responses arise from the neurons of the nucleus reticularis thalami, some distance away from the site of recording and iontophoretic drug application, it is likely that the reduction of inhibition seen with L-AP4 and CCG1 is due to an action of these agonists on the terminals or axons of these inhibitory neurons. The novel antagonists of L-AP4 and CCG1, alpha-methyl-L-AP4 and alpha-methyl-CCG1, were found to block the disinhibitory actions of the agonists in a differential manner when applied iontophoretically. This suggests that there may be at least two types of receptor mediating the disinhibitory effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Increased messenger RNA expression of the 695 and 751 amino acid isoforms of the beta-amyloid protein precursor in the thalamus of 17-year-old cynomolgus (Macaca fascicularis) monkeys.

The levels of expression of messenger RNAs of the 695 and 751 amino acid isoforms of the beta-amyloid protein precursor in the brains of three-year-old and 17-year-old cynomolgus monkeys (Macaca fascicularis) were visualized and quantified by in situ hybridization histochemistry using 35S-labelled oligonucleotide probes. The analysis was carried out on coronal brain sections taken through the hippocampus and thalamus at the level of the geniculate nuclei. High densities of beta-amyloid protein precursor695 and beta-amyloid protein precursor751 messenger RNAs were found in the medial aspects of the mediodorsal, centromedian and parafascicular nuclei of the 17-year-old monkeys. The messenger RNA levels of the 695 and 751 isoforms were about two- and seven-fold, respectively, those found in the same nuclei of the three-year-old animals. The levels of these messenger RNA transcripts in the 17-year-old monkeys were not significantly different from those in the three-year-old animals in other brain areas e.g. the temporal cortex, entorhinal cortex and hippocampus. No Alzheimer's disease-like neuropathology in terms of diffuse or senile beta-amyloid plaques, dystrophic neurites or neurofibrillary tangles were detectable by specific innumohistochemical procedures in the above thalamic nuclei of the 17-year-old animals. In addition no reactive gliosis was seen in the thalamus of these monkeys.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1994

Fine structural alteration in target-deprived axonal terminals in the rat thalamus.

Lesioning of thalamic target neurons in the adult rat provokes a resorption of terminal axonal branches of afferent neurons from the dorsal column nuclei. There is, however, no massive neuronal loss in the dorsal column nuclei. In the adult, therefore, the thalamic post synaptic target cells influence primarily the extent of the terminal component of the afferent neurons. The subcellular changes underlying the regression of these adult terminals are unknown. To address this issue, we have looked at the electron microscopic level for the ultrastructural correlates of this retraction of the terminal compartment of target-deprived neurons in the adult rat thalamus. By analysing the fine structure of target-deprived axons and their immunoreactivity for a specific synaptic protein, synaptophysin, we have observed that all the organelles and the protein formed in the cell body, including dense-core vesicles, continue to be transported to the terminal compartment and accumulate at this level. At the terminal level, engorgement of organelles induces the formation of varicosities. An enormous increase in local degradative activity occurs in parallel to this accumulation. In contrast, organelles involved in membrane turnover and degradation (including synaptic and coated vesicles, multivesicular bodies, lysosomes) in the nerve terminals are clearly modified. There is a progressive loss of synaptic vesicles, whereas clathrin-coated vesicles and multivesicular bodies are numerous. We propose that the resorption of terminal axonal branches after thalamic target deprivation in the adult is associated with a bias of the system of membrane recycling at the axonal terminals towards degradation. In the absence of apparent changes in the pathways originating from the cell body in these conditions, it is unlikely that presynaptic neurons in the adult thalamus are dependent upon post synaptic target neurons for the delivery of organelles and proteins to the terminals.

Animals↗
PubMed1994

Growth-promoting interactions between the murine neocortex and thalamus in organotypic co-cultures.

The aim of this study was to assess whether developing cerebral cortex produces diffusible factors that can affect the growth of thalamic cells and, if so, what the role of these factors might be during the formation of thalamocortical connections. We studied interactions between cultured organotypic explants from mice maintained in defined serum-free medium. First, we cultured explants of embryonic dorsolateral thalamus in isolation from any other tissue; after culture, these explants were viewed intact and then sectioned. We estimated the numbers of healthy and pyknotic cells before and after culture, and the rates of mitosis in the explants during culture (using bromodeoxyuridine). Based on these data, we concluded that the majority of cells in the thalamic explants survived, although significant numbers of pyknotic cells did accumulate. Thalamic explants extended either very few or no neurites when cultured alone. We then cultured explants of embryonic thalamus near to explants from other tissues. A gap was always maintained between the explants, and we measured the length and density of neurite outgrowth from each thalamic explant. Slices of embryonic cortex promoted a small but significant increase in the amount of growth from thalamic explants. Postnatal cortex stimulated much more profuse neurite outgrowth; postnatal cerebellum had less of an effect, and postnatal medulla or liver had none. We showed that there was significantly more outgrowth from thalamic explants cultured in medium that had been preconditioned with cortical slices than from thalamic explants cultured in control medium, confirming that diffusible factors were produced by the cortex. The survival and mitotic rates of thalamic cells were unaffected by co-culture with the cortex. We conclude that the developing cortex releases diffusible factors that stimulate the growth of thalamic neurites and that other regions of the brain may also release the same substance(s). The lack of a specific source of thalamic growth promoting factor(s) argues against a role for these factors in guiding thalamic axons to specific targets; indeed, we were unable to demonstrate any chemotropic guidance of thalamic axons towards cortical explants in collagen gels. Since postnatal cortex has a more potent stimulatory effect than prenatal cortex, it seems possible that, in vivo, the cortical-derived factors act mainly on thalamocortical axons that have located their targets and are in the process of arborizing and refining their connections.

Animals↗
PubMed1982

Intracranial self-stimulation in the thalamus of the rat.

Rats were tested for intracranial self-stimulation (ICSS) via bipolar electrodes situated throughout the thalamus. Of 112 animals in the study, 55 met the criteria for ICSS, with scores ranging from 55 to 921 bar-presses in a 15 minute session. A map of both positive and neutral placements is presented. Positive sites for ICSS were found in all aspects of the mediodorsal nucleus, except for the central segment. The ventromedial nuclear complex was also a positive area of ICSS, with the exception of the submedial nucleus (nucleus gelatinosus). Each of the intralaminar nuclei (central medial, parafascicular, paracentral, and central lateral) supported ICSS, as did each of the midline nuclei (rhomboid, paratenial and paraventricular). No placements were found in the nucleus reuniens. Both "major" relay nuclei, the ventrobasal and ventrolateral, supported ICSS; but neither the laterodorsal nor the lateral posterior nuclei had positive caudal thalamus. As a general rule, ICSS scores appeared to be higher as the electrode placements approached the midline. Sites in which no positive placements were seen included the reticular nucleus, as well as the stria medularis, the mammillothalamic tract, and the fasciculus retroflexus.

Animals↗
PubMed1988

Differential effects on locomotor activity of injections of procaine into mediodorsal thalamus and pedunculopontine nucleus.

A comparison was made of the contributions to locomotor activity of output projections of the subpallidal region to the mediodorsal thalamus (MD) and to the pedunculopontine nucleus (PPN). Locomotor activity elicited by injections of picrotoxin into the subpallidal region was reduced by the administration of procaine to the pedunculopontine nucleus but not by the administration of procaine to mediodorsal thalamus. Since the pedunculopontine nucleus is part of the mesencephalic locomotor region (MLR) it appears that subpallido-pedunculopontine projections contribute to the locomotor component of adaptive behaviors associated with limbic integrative activities.

Animals↗
PubMedDate not supplied

Immunocytochemical study of enkephalin-like cell bodies in the thalamus of the rat.

The distribution of enkephalin-like cell bodies in the thalamus of the rat was studied by means of intratissular injections of colchicine and using an indirect immunoperoxidase technique. The densest clusters of immunoreactive perikarya were observed in the nuclei geniculatum lateralis ventralis, medialis dorsalis, centralis lateralis, centralis medialis and anterior ventralis. Whereas the nuclei praetectalis lateralis, lateralis posterior, habenularis lateralis, parataenialis (its caudal part), parafascicularis, centrum medianum, reuniens and ventralis medialis had the lowest density. In other thalamic nuclei geniculatum mediale, paraventricularis and parataenialis (its rostral part) the density of enkephalin-like cell bodies was intermediate. These results suggest that the intratissular injection of colchicine is the better way of administration of the drug in order to study the distribution of peptidergic cell bodies in the mammalian CNS. The similarities and differences found in the distribution of enkephalinergic cell populations in the thalamus of different mammals are discussed.

Animals↗
PubMed1991

Retinofugal projections to the hypothalamus, anterior thalamus and basal forebrain in hamsters.

In Part a of the study, the retinal inputs to the hypothalamus, anterior thalamus and basal forebrain of Syrian hamsters were studied using intraocular injections of horseradish peroxidase conjugated to cholera toxin (CT-HRP). In the hypothalamus, the heaviest retinal input was to the suprachiasmatic nucleus (SCN), however, many labeled fibers coursed through the SCN to reach more caudal, periventricular and lateral sites including the anterior and lateral hypothalamus, the paraventricular nucleus (PVN), the subparaventricular zone, the ventromedial nucleus and the pars compacta of the dorsomedial nucleus. Some of these fibers continued dorsally into the zona incerta (ZI). Other fibers emerged from the lateral optic chiasm and traveled either rostro-medially to end in the preoptic area (POA) or further laterally to reach the supraoptic nucleus. A subset of fibers extended laterally from the chiasm to form a well-defined tract which provided input to the pyriform cortex. The extrageniculate retinal input to the thalamus was to the anterior thalamic area (AT) via the stria terminalis. In Part b, injections of rhodamine-labeled latex microspheres were made in three brain areas that contained labeled fibers after intraocular injections of CT-HRP. Injections in the AT, PVN/ZI area and POA consistently produced a small number of labeled retinal ganglion cells, whereas control injections did not. Taken together, these results indicate that many regions of the brain involved in the control of reproductive and regulatory functions receive photic informations via direct retinal inputs. These retinal inputs may play a role in the photoperiodic modulation of physiology and behavior.

Animals↗
PubMed1994

Distribution of calretinin, calbindin-D28k, and parvalbumin in the rat thalamus.

The localization of three calcium-binding proteins, calretinin, calbindin-D28k, and parvalbumin, in the rat thalamus was immunohistochemically examined. a) Some thalamic regions revealed cells almost exclusively containing one of the calcium-binding proteins. For example, almost only calretinin-stained cells were found in the central medial and paraventricular nuclei. Calbindin-D28k-stained cells were mostly found in the centrolateral, interanteromedial, anteromedial, and posterior nuclei. Only parvalbumin-positive cells were found in the central part of the reticular nucleus. b) Other regions expressed overlap between the distributions of two cell components composed of different calcium-binding proteins. For example, both calretinin-stained cells and calbindin-D28k-labeled cells were found in the lateroposterior, intermediodorsal, rhomboid, and reuniens nuclei. c) Other regions showed no cells stained for any of the calcium-binding proteins. For example, generally no calcium-binding protein was detected in neurons of the anterodorsal, anteroventral, ventrolateral, ventral posterolateral, ventral posteromedial, or gelatinosus nuclei, or of the central part of the mediodorsal nucleus. These three proteins serve as useful marker for localizing subpopulations of neurons within the thalamus.

Animals↗
PubMed1994

Afferent connections of the anterior thalamus in rabbits.

This study was designed to determine whether axons of cholinergic dorsal tegmental neurons terminate on cells in the anterior thalamus in rabbits as in other species, and to localize projecting tegmental cells for future studies of their contributions to anterior thalamic learning-relevant neuronal activity. The distribution of retrogradely labeled neurons was examined following injections of wheat germ agglutinin horseradish peroxidase (WGA-HRP) centered in the anterior ventral (AV) thalamic nucleus. The results confirm past findings in rabbits indicating projections to anterior thalamus from the mammillary nuclei, the posterior cingulate cortex, presubiculum and postsubiculum. Demonstrated for the first time in rabbits were projections from the lateral dorsal and the pedunculopontine tegmental nuclei, locus coeruleus, dorsal raphe nucleus, Gudden's dorsal tegmental nucleus, pretectum and reticular thalamic nucleus.

Afferent Pathways↗
PubMed1993

Migration pathways, differentiation and survival of macroglial cells from a xenograft implanted into the thalamus of newborn mice.

Embryonic rabbit corpus callosum transplants were grafted into thalamus of newborn shiverer mice in order to compare the fates of oligodendroglial and astroglial cells derived from the transplants. Our model allowed the identification of the two populations of macroglial cells. The thalamus was chosen as site of implantation because of its situation at a crossroad of numerous neuronal fascicles. Previous studies, where the dorsal striatum was used as site of implantation, had shown that corpus callosum was one of the favorite routes of migration for both populations of macroglial cells. In the present study special attention was given to the comparison of the migration pathways and areas of settlement of implanted astroglia and oligodendroglia. The internal capsule, the medial lemniscus, the crus cerebri and the thalamic radiations were used by both populations of transplant derived macroglial cells for their migrations through the host parenchyma. They integrated into the host tissue on these routes or further away in areas such as the putamen, the mesencephalon or the colliculi. Signs of degeneration of the implanted astroglia were often observed after 1 month post-implantation.

Animals↗
PubMedDate not supplied

Ultrasonographic findings in thalamus and basal ganglia in term asphyxiated infants.

Three severely asphyxiated term neonates demonstrated bilateral hyperechogenicity in the thalamus and basal ganglia. During evolution, areas of attenuated echogenicity appeared in these structures at the same time as periventricular cysts were evident in 2 patients with coexistent periventricular leukomalacia. All 3 patients developed ventricular dilatation; in the 2 patients with periventricular leukomalacia, the ventricular border was irregular in the outer (dorsal) margin, and interhemispheric fissures were widened as a manifestation of cerebral atrophy. Furthermore, the thalamic inner (ventral) margins of the lateral ventricles were irregular in all 3 patients. This previously unrecognized finding points to a particular form of cerebral atrophy localized in the gangliothalamic region that contributes to the development of ventriculomegaly. The reported sonographic sequence implies profound damage in the thalamus and basal ganglia in asphyxiated infants which undoubtedly has contributed to the poor outcomes of our patients.

Asphyxia Neonatorum↗
PubMed1994

New pattern of hyperechogenicity in thalamus and basal ganglia studied by color Doppler flow imaging.

Thirty-seven infants whose cerebral real-time B-mode ultrasound (CUS) documented hyperechogenic areas in the thalamus and basal ganglia (HTBG) either of linear or fine punctate pattern, were studied prospectively by color Doppler imaging (CDI). This study aimed to establish a relationship between these areas and the regional vasculature, to analyze associated disorders to establish pathogenesis, and to determine clinical significance. HTBG were diagnosed in the first 4 days of life in all but 7 infants. Different patterns of HTBG were observed: punctate in 11 infants, linear in 12, and mixed in 14. The basal ganglia were affected in all patients, 9 also had involvement of the thalamus. CDI confirmed that HTBG were allocated along the gangliothalamic vessels. Blood flow velocity waves were obtained at this level in all patients. Real-time spectral analyses were performed in 35 patients and compared with a control group of 20 healthy neonates. Differences were not significant. Computed tomography and magnetic resonance imaging failed to indicate this abnormality. Necropsy revealed basophilic deposits in the walls of involved arteries. Congenital infections manifested in 5 patients, chromosomal abnormality in 1, dysmorphic syndromes in 9 (3 unidentified), isolated congenital defects in 5, and diverse congenital disorders in 3. In the remaining 14, no congenital disorders nor infections were found. This CDI study demonstrates the vascular location of these HTBG. Supported by early CUS diagnosis, it is speculated that vascular injury in that region has a prenatal origin. This abnormality does not appear to alter regional blood flow. HTBG are associated with very heterogeneous disorders and in most patients the etiology and pathogenesis remain unclear.

Arteries↗
PubMedDate not supplied

The relationship of calbindin-containing neurons with substance P, Leu-enkephalin and cholecystokinin fibres: an immunohistochemical study in the rat thalamus.

In the rat thalamus, immunoreactivity for the calcium binding protein calbindin (Cb) is mostly confined to neuronal cell bodies, sometimes revealing proximal dendrites, of the midline, intralaminar and posterior regions. Substance P (SP)-, cholecystokinin (CCK)- and Leu-enkephalin (L-ENK)-immunoreactive (ir) elements in the thalamus are fibre-like structures, intermingled with punctate elements probably representing axonal arborizations and their synaptic boutons. These peptidergic fibres are unevenly distributed in several thalamic domains, including the areas that contain Cb-ir neurons. The relationship between Cb-ir cell bodies and these three different peptidergic systems of thalamic innervation was studied with immunohistochemistry. Single-labelling experiments on adjacent sections and double immunostaining on the same section were performed. A considerable overlap between Cb-ir perikarya and SP-ir fibres was found in most thalamic nuclei. In particular, in the intralaminar nuclei and posterior complex, SP-ir punctate elements were frequently observed in close proximity to Cb-ir cell bodies and dendrites. On the other hand, no consistent topographical correspondence between Cb-ir perikarya and CCK- or L-ENK-ir fibres was evident. Altogether, the present data suggest a selective anatomical and, possibly, functional relationship between SP and Cb in at least a subpopulation of rat thalamic neurons.

Animals↗