The current study describes, using immunohistochemical methods, the nuclear organization of the cholinergic, catecholaminergic and serotonergic systems within the brains of five microchiropteran species. For the vast majority of nuclei observed, direct homologies are evident in other mammalian species; however, there were several distinctions in the presence or absence of specific nuclei that provide important clues regarding the use of the brain in the analysis of chiropteran phylogenetic affinities. Within the five species studied, three specific differences (presence of a parabigeminal nucleus, dorsal caudal nucleus of the ventral tegmental area and the absence of the substantia nigra ventral) found in two species from two different families (Cardioderma cor; Megadermatidae, and Coleura afra; Emballonuridae), illustrates the diversity of microchiropteran phylogeny and the usefulness of brain characters in phylogenetic reconstruction. A number of distinct differences separate the microchiropterans from the megachiropterans, supporting the diphyletic hypothesis of chiropteran phylogenetic origins. These differences phylogenetically align the microchiropterans with the heterogenous grouping of insectivores, in contrast to the alignment of megachiropterans with primates. The consistency of the changes and stasis of neural characters with mammalian phylogeny indicate that the investigation of the microchiropterans as a sister group to one of the five orders of insectivores to be a potentially fruitful area of future research.

Studies detailing the anatomy of the brain of the golden moles are few. A recent study indicated that in the Hottentot golden mole (a member of the Amblysominae clade), there was a broad, atypical, distribution of cholinergic interneurons in the olfactory bulb, cerebral cortex, hippocampus and amygdala. To determine whether this broad distribution of cholinergic neurons is shared by other species of golden mole, we here examine the brain of the Cape golden mole (a member of the Chrysochlorinae clade, representing the second major clade within the family Chrysochloridae). Our analyses indicates the presence of a similar widespread distribution of cholinergic interneurons in the Cape golden mole. Thus, we conclude that these features are derived morphological traits in the brains of golden moles. In addition, we describe the nuclei generally considered to be part of the typical cholinergic system in mammals. Whereas the vast majority of these generally reported cholinergic nuclei were the same as recorded in other Eutherian mammals, it was noted that the cholinergic nuclei involved in oculomotion were substantially reduced in size, or absent in the case of the abducens nucleus. In addition, there was an absence of the cholinergic medial septal nucleus, but the presence of a cholinergic lateral septal nucleus. The laterodorsal and pedunculopontine tegmental nuclei evince regions where the cholinergic neurons are densely packed. These are atypical features of the mammalian cholinergic system, which when combined with the widespread atypical distribution of cholinergic interneurons, reveals a family-specific complement of cholinergic nuclei in the Chrysochloridae.

The nuclear organization of the cholinergic, catecholaminergic, serotonergic and orexinergic neurons in the brains of two species of carnivore, the banded mongoose (Mungos mungo) and domestic ferret (Mustela putorius furo), is presented. The banded mongoose belongs to the feliform suborder and the domestic ferret to the caniform suborder, having last shared a common ancestor approximately 53 million years ago; however, they have a very similar overall morphology and life history, presenting an interesting opportunity to examine the extent of evolutionary plasticity in these systems. The brains of the two carnivore species were coronally sectioned and immunohistochemically stained with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. The overall organization and complement of the nuclei of these systems was identical between the two species, although minor differences were noted. Moreover, this overall organization is identical to other studies undertaken in the domestic cat and dog. While for the most part the nuclei forming these systems are similar to those observed in other mammals, two species differences, which appear to be carnivore-specific, were noted. First, cholinergic neurons were observed in the lateral septal nucleus of both species, an apparently carnivore specific feature not recorded previously in other mammals. Second, the serotonergic neurons of the peripheral division of the dorsal raphe complex exhibited a significant caudad expansion, intermingling with the cholinergic and catecholaminergic nuclei of the pons, a carnivore specific feature. These carnivore specific features likely have functional consequences related to coping with stress and the expression of sleep.

Here we report the unusual presence of thalamic reticular neurons immunoreactive for tyrosine hydroxylase in equids. The diencephalons of one adult male of four equid species, domestic donkey (Equus africanus asinus), domestic horse (Equus caballus), Cape mountain zebra (Equus zebra zebra) and plains zebra (Equus quagga), were sectioned in a coronal plane with series of sections stained for Nissl substance, myelin, or immunostained for tyrosine hydroxylase, and the calcium-binding proteins parvalbumin, calbindin and calretinin. In all equid species studied the thalamic reticular nucleus was observed as a sheet of neurons surrounding the rostral, lateral and ventral portions of the nuclear mass of the dorsal thalamus. In addition, these thalamic reticular neurons were immunopositive for parvalbumin, but immunonegative for calbindin and calretinin. Moreover, the thalamic reticular neurons in the equids studied were also immunopositive for tyrosine hydroxylase. Throughout the grey matter of the dorsal thalamus a terminal network also immunoreactive for tyrosine hydroxylase was present. Thus, the equid thalamic reticular neurons appear to provide a direct and novel potentially catecholaminergic innervation of the thalamic relay neurons. This finding is discussed in relation to the function of the thalamic reticular nucleus and the possible effect of a potentially novel catecholaminergic pathway on the neural activity of the thalamocortical loop.

Children with Fetal Alcohol Spectrum Disorder (FASD) have impaired sensory processing skills as a result of neurodevelopmental anomalies. The somatosensory barrel field of rodent brain is a readily accessible model for studying the effects of alcohol exposure. Within the barrel field, the posterior medial barrel subfield (PMBSF) receives sensory inputs from the large vibrissae on the contralateral face. This study reports on the consequence of prenatal exposure to alcohol on the somatosensory cortices of mice later in life. Two control groups, a sucrose and a non-treated control, were also examined. At postnatal day (PND) 56 the cerebral hemisphere of mice from each group were processed for cytochrome oxidase reactivity. In contrast to previous studies, there were no significant differences in the mean areas of: (I) the PMBSF enclosure, (II) the PMBSF barrels, (III) the individual PMBSF barrels and (IV) the septal portion of the PMBSF in the alcohol group compared to the controls. However barrel sizes in rows D and E in the alcohol group were significantly reduced, indicating an alcohol-induced damage on the barrel development and which may reduce the amount of the cortex devoted to processing somatosensory input- a common defect seen in children with FASD.

The present study describes the distribution of Orexin-A immunoreactive cell bodies and terminal networks in the brains of five microchiropteran species. Given the specialized flight and echolocation abilities of the microchiropterans it was of interest to examine if any specific differences in a generally phylogenetically homogenous neural system could be found. The orexinergic neurons have been found within the hypothalamus of all species studied, and were represented by a large cluster that spanned the anterior, dorsomedial, perifornical and lateral hypothalamic regions, with a smaller cluster extending into the region of the medial zona incerta. Evidence for orexinergic neurons in the ventrolateral hypothalamus adjacent to the optic tract was not observed in any microchiropteran species. The terminal networks of the orexinergic neurons conformed to that previously reported in a range of mammalian species, with dense terminal networks being found in the hypothalamus, cholinergic pedunculopontine and laterodorsal tegemental nuclei, the noradrenergic locus coeruleus complex, all serotonergic nuclei, the paraventricular nuclei of the epithalamus and adjacent to the habenular nuclei. Thus, apart from the lack of neurons in the ventrolateral hypothalamus, the orexinergic system of the microchiropterans appears typically mammalian.

This study investigated the nuclear organization of four immunohistochemically identifiable neural systems (cholinergic, catecholaminergic, serotonergic and orexinergic) within the brains of three male Tasmanian devils (Sarcophilus harrisii), which had a mean brain mass of 11.6g. We found that the nuclei generally observed for these systems in other mammalian brains were present in the brain of the Tasmanian devil. Despite this, specific differences in the nuclear organization of the cholinergic, catecholaminergic and serotonergic systems appear to carry a phylogenetic signal. In the cholinergic system, only the dorsal hypothalamic cholinergic nucleus could be observed, while an extra dorsal subdivision of the laterodorsal tegmental nucleus and cholinergic neurons within the gelatinous layer of the caudal spinal trigeminal nucleus were observed. Within the catecholaminergic system the A4 nucleus of the locus coeruleus complex was absent, as was the caudal ventrolateral serotonergic group of the serotonergic system. The organization of the orexinergic system was similar to that seen in many mammals previously studied. Overall, while showing strong similarities to the organization of these systems in other mammals, the specific differences observed in the Tasmanian devil reveal either order specific, or class specific, features of these systems. Further studies will reveal the extent of change in the nuclear organization of these systems in marsupials and how these potential changes may affect functionality.

The present study describes the distribution of orexin-A immunoreactive neurons and their terminal networks in the brains of two species of megachiropterans. In general the organization of the orexinergic system in the mammalian brain is conserved across species, but as one of two groups of mammals that fly and have a high metabolic rate, it was of interest to determine whether there were any specific differences in the organization of this system in the megachiropterans. Orexinergic neurons were limited in distribution to the hypothalamus, and formed three distinct clusters, or nuclei, a main cluster with a perifornical location, a zona incerta cluster in the dorsolateral hypothalamus and an optic tract cluster in the ventrolateral hypothalamus. The nuclear parcellation of the orexinergic system in the megachiropterans is similar to that seen in many mammals, but differs from the microchiropterans where the optic tract cluster is absent. The terminal networks of the orexinergic neurons in the megachiropterans was similar to that seen in a range of mammalian species, with significant terminal networks being found in the hypothalamus, cholinergic pedunculopontine and laterodorsal tegemental nuclei, the noradrenergic locus coeruleus complex, all serotonergic nuclei, the paraventricular nuclei of the epithalamus and adjacent to the habenular nuclei. While the megachiropteran orexinergic system is typically mammalian in form, it does differ from that reported for microchiropterans, and thus provides an additional neural character arguing for independent evolution of these two chiropteran suborders.

The present study describes the organization of the orexinergic (hypocretinergic) neurons in the hypothalamus of the giraffe and harbour porpoise--two members of the mammalian Order Cetartiodactyla which is comprised of the even-toed ungulates and the cetaceans as they share a monophyletic ancestry. Diencephalons from two sub-adult male giraffes and two adult male harbour porpoises were coronally sectioned and immunohistochemically stained for orexin-A. The staining revealed that the orexinergic neurons could be readily divided into two distinct neuronal types based on somal volume, area and length, these being the parvocellular and magnocellular orexin-A immunopositive (OxA+) groups. The magnocellular group could be further subdivided, on topological grounds, into three distinct clusters--a main cluster in the perifornical and lateral hypothalamus, a cluster associated with the zona incerta and a cluster associated with the optic tract. The parvocellular neurons were found in the medial hypothalamus, but could not be subdivided, rather they form a topologically amorphous cluster. The parvocellular cluster appears to be unique to the Cetartiodactyla as these neurons have not been described in other mammals to date, while the magnocellular nuclei appear to be homologous to similar nuclei described in other mammals. The overall size of both the parvocellular and magnocellular neurons (based on somal volume, area and length) were larger in the giraffe than the harbour porpoise, but the harbour porpoise had a higher number of both parvocellular and magnocellular orexinergic neurons than the giraffe despite both having a similar brain mass. The higher number of both parvocellular and magnocellular orexinergic neurons in the harbour porpoise may relate to the unusual sleep mechanisms in the cetaceans.

The nuclear organization of the cholinergic, putative catecholaminergic and serotonergic systems within the brains of the megachiropteran straw-coloured fruit bat (Eidolon helvum) and Wahlberg's epauletted fruit bat (Epomophorus wahlbergi) were identified following immunohistochemistry for cholineacetyltransferase, tyrosine hydroxylase and serotonin. The aim of the present study was to investigate possible differences in the nuclear complement of the neuromodulatory systems of these species in comparison to previous studies on megachiropterans, microchiropterans and other mammals. The nuclear organization of these systems is identical to that described previously for megachiropterans and shows many similarities to other mammalian species, especially primates; for example, the putative catecholaminergic system in both species presented a very compact nucleus within the locus coeruleus (A6c) which is found only in megachiropterans and primates. A cladistic analysis of 38 mammalian species and 82 characters from these systems show that megachiropterans form a sister group with primates to the exclusion of other mammals, including microchiropterans. Moreover, the results indicate that megachiropterans and microchiropterans have no clear phylogenetic relationship to each other, as the microchiropteran systems are most closely associated with insectivores. Thus a diphyletic origin of Chiroptera is supported by the present neural findings.