Human origins and evolution: Cold Spring Harbor, deja vu.

The Cold Spring Harbor Symposia of the 1950s were key to integrating human evolutionary studies into biology. That integration provided a solid foundation for systematic and functional interpretations of an expanding base of fossil and molecular evidence during the latter half of the 20th century. Today, the paleontological record of human evolution amassed during the last 150 years illuminates the human clade on life's tree. However, the rise of Hennegian parsimony cladistics and punctuationalism during the end of the last century witnessed the partial abandonment of classificatory conventions cemented by Mayr, Simpson, Dobzhansky, and others at Cold Spring Harbor. This has led to an artificial, postmillennial amplification of apparent species diversity in the hominid clade. Work on a stratigraphically thick and temporally deep sedimentary sequence in the Middle Awash study area of Ethiopia's Afar Depression reveals an assembly order of hominid anatomies and behaviors that was impossible for Darwin to discern. Large parts of that record appear to reflect phyletic evolution, consistent with the lessons and expectations of Cold Spring Harbor in 1950. Molecular biology cannot reveal the assembly sequences or contexts of human origins and evolution without reference to adequate geological, geochronological, paleobiological, and archaeological records. Today's consilience of these disparate data sets would have impressed Charles Darwin.

Hominid cranial remains from upper Pleistocene deposits at Aduma, Middle Awash, Ethiopia.

The Upper Pleistocene localities of Aduma and Bouri have yielded hominid fossils and extensive Middle Stone Age (MSA) archaeological assemblages. The vertebrate fossils recovered include parts of four hominid crania from Aduma and a complete right parietal from Bouri. Archaeological associations and radiometric techniques suggest an Upper Pleistocene age for these hominids. The more complete cranium from Aduma (ADU-VP-1/3) comprises most of the parietals, the occipital, and part of the frontal. This cranium is compared to late Middle and Upper Pleistocene hominid crania from Africa and the Middle East. The Aduma cranium shows a mosaic of cranial features shared with "premodern" and anatomically modern Homo sapiens. However, the posterior and lateral cranial dimensions, and most of its anatomy, are centered among modern humans and resemble specimens from Omo, Skhul, and Qafzeh. As a result, the Aduma and Bouri Upper Pleistocene hominids are assigned to anatomically modern Homo sapiens.

Activation of cholinergic and adrenergic receptors increases the concentration of extracellular adenosine in the cerebral cortex of unanesthetized rat.

Adenosine is an inhibitory neuromodulator in the CNS. For extracellular adenosine to play a physiological role in the brain, it must be present at effective concentrations. Acetylcholine and noradrenaline are known to play an important role in modulating the activity of cortical neurons, and they might have a role also in the release of adenosine in the cerebral cortex in vivo. We examined whether activation of cholinergic and adrenergic receptors affects extracellular adenosine levels in the cerebral cortex of unanesthetized rats using in vivo microdialysis. All drugs were administered locally within the cortex by reverse dialysis. Both acetylcholine and the acetylcholinesterase inhibitor neostigmine increased extracellular adenosine levels, and the effect of neostigmine was blocked by the nicotinic receptor antagonist mecamylamine. Both nicotine and the nicotinic receptor agonist epibatidine increased the concentration of extracellular adenosine. Activation of muscarinic receptors using the nonselective agonist oxotremorine and a selective M1 receptor agonist also increased extracellular adenosine levels. Noradrenaline and the noradrenergic reuptake inhibitor desipramine increased extracellular adenosine levels. The alpha(1)-adrenergic receptor agonist phenylephrine and the beta-adrenergic agonist isoproterenol increased extracellular adenosine levels, whereas the alpha(2)-adrenergic receptor agonist clonidine did not have an effect. These findings indicate that activation of specific cholinergic and adrenergic receptors can increase extracellular levels of adenosine in the cortex, and suggest that cholinergic and adrenergic receptor-mediated regulation of adenosine levels may represent a mechanism for controlling the excitability of cortical neurons.

Enhanced release of adenosine in rat hind paw following spinal nerve ligation: involvement of capsaicin-sensitive sensory afferents.

Modulation of endogenous adenosine levels by inhibition of adenosine metabolism produces a peripheral antinociceptive effect in a neuropathic pain model. The present study used microdialysis to investigate the neuronal mechanisms modulating extracellular adenosine levels in the rat hind paw following tight ligation of the L5 and L6 spinal nerves. Subcutaneous injection of 50 microl saline into the nerve-injured paw induced a rapid and short-lasting increase in extracellular adenosine levels in the subcutaneous tissues of the rat hind paw ipsilateral to the nerve injury. Saline injection did not increase adenosine levels in sham-operated rats or non-treated rats. The adenosine kinase inhibitor 5'-amino-5'-deoxyadenosine and the adenosine deaminase inhibitor 2'-deoxycoformycin, at doses producing a peripheral antinociceptive effect, did not further enhance subcutaneous adenosine levels in the nerve-injured paw. Systemic pretreatment with capsaicin, a neurotoxin selective for small-diameter sensory afferents, markedly reduced the saline-evoked release of adenosine in rat hind paw following spinal nerve ligation. Systemic pretreatment with 6-hydroxydopamine, a neurotoxin selective for sympathetic afferent nerves, did not affect release. These results suggest that following nerve injury, peripheral capsaicin-sensitive primary sensory afferent nerve terminals are hypersensitive, and are able to release adenosine following a stimulus that does not normally evoke release in sham-operated or intact rats. Sympathetic postganglionic afferents do not appear to be involved in such release. The lack of effect on such release by the inhibitors of adenosine metabolism suggests an altered peripheral adenosine system following spinal nerve ligation.

Involvement of primary sensory afferents, postganglionic sympathetic nerves and mast cells in the formalin-evoked peripheral release of adenosine.

Injection of formalin into the rat hind paw produces a dose-dependent local peripheral release of adenosine. Low doses of formalin (0.5-2.5%) evoke release during the first 10 min following injection, while a high dose of formalin (5%) evokes release lasting for 60 min. The current study was designed to determine the possible origin of release produced by two doses of formalin (1.5% and 5%). Microdialysis probes were implanted into the subcutaneous tissue under the glabrous skin of the hind paw of anaesthetized rats, and adenosine was determined by high performance liquid chromatography. Pretreatment with capsaicin, a neurotoxin selective for unmyelinated small diameter primary afferent nerves, markedly reduced the adenosine released by 1.5% formalin and the early phase of release by 5% formalin. Acute injection of 1% capsaicin to the hind paw of untreated rats also induced adenosine release. Pretreatment with 6-hydroxydopamine, a neurotoxin selective for sympathetic postganglionic nerve terminals, had no effect on release evoked by 1.5% formalin, but significantly reduced adenosine release during the late phase of release induced by 5% formalin. Pretreatment with compound 48/80, which degranulates mast cells, had no effect on adenosine release evoked by either concentration of formalin. We conclude that the origin of the adenosine released peripherally by formalin depends on the formalin concentration. At the lower concentration (1.5%), release is predominantly from unmyelinated sensory afferent nerve terminals, while at the higher concentration (5%), unmyelinated afferent nerve terminals are involved in the early phase, while sympathetic postganglionic nerve terminals are involved in the later phase. Mast cells do not contribute to release of adenosine evoked by either concentration of formalin.

Prey processing in amniotes: biomechanical and behavioral patterns of food reduction.

In this paper we examine the biomechanics of prey processing behavior in the amniotes. Whether amniotes swallow prey items whole or swallow highly processed slurries or boluses of food, they share a common biomechanical system where hard surfaces (teeth or beaks) are brought together on articulated jaws by the actions of adductor muscles to grasp and process food. How have amniotes modified this basic system to increase the chewing efficiency of the system? To address this question we first examine the primitive condition for prey processing representative of many of the past and present predatory amniotes. Because herbivory is expected to be related to improved prey processing in the jaws we review patterns of food processing mechanics in past and present herbivores. Herbivory has appeared numerous times in amniotes and several solutions to the task of chewing plant matter have appeared. Birds have abandoned jaw chewing in favor of a new way to chew--with the gut--so we will detour from the jaws to examine the appearance of gut chewing in the archosaurs. We will then fill in the gaps among amniote taxa with a look at some new data on patterns of prey processing behavior and jaw mechanics in lizards. Finally, we examine evolutionary patterns of amniote feeding mechanism and how correlates of chewing relate to the need to increase the efficiency of prey processing in order to facilitate increased metabolic rate and activity.

Activation of metabotropic glutamate receptors increases extracellular adenosine in vivo.

In order to identify the mechanisms that would lead to increased levels of the inhibitory neuromodulator adenosine in the brain, we tested metabotropic glutamate receptor agonists for their ability to increase extracellular adenosine in the cortex of unanesthetized rat using in vivo microdialysis. The group I/II metabotropic glutamate receptor agonist trans-(+/-)- 1-amino-(1S,3R)-cyclopenyanedicarboxylic acid (I mM) increased extracellular adenosine as did the specific group I agonist (S)- 3,5-dihydroxyphenylglycine (DHPG; 1 mM). The evoked increase of adenosine by 1 mM DHPG was reduced by the group I antagonist (RS)- 1-aminoindan-1,5,-dicarboxylic acid. Activation of group II or III metabotropic receptors did not affect extracellular adenosine. These results suggest that activation of group I metabotropic receptors contributes to elevated extracellular adenosine levels in vivo.

Potentiation of formalin-evoked adenosine release by an adenosine kinase inhibitor and an adenosine deaminase inhibitor in the rat hind paw: a microdialysis study.

The present study examined the effects of local subcutaneous administration of formalin on adenosine release from the rat hind paw, and the effects of inhibitors of adenosine metabolism on such release. Microdialysis probes were inserted into the subcutaneous tissue of the plantar surface of rat hind paws. Samples were collected every 10 min at a perfusion rate of 2 microl/min and high performance liquid chromatography was used to measure adenosine levels. At lower concentrations of formalin (0.5-2.5%), a significant increase in adenosine levels was observed in the first 10 min after formalin injection, while at the highest concentration of formalin (5%), the increase in adenosine release was observed over 60 min. Co-administration of the adenosine kinase inhibitor 5'-amino-5'-deoxyadenosine (100 nmol) with formalin, significantly increased adenosine release evoked by 0.5-1.5% formalin, but did not produce a further enhancement of release evoked by 5% formalin. The adenosine deaminase inhibitor 2'-deoxycoformycin (100 nmol) significantly increased adenosine levels at 5% formalin but had no effect at lower concentrations of formalin. In confirmation of previous studies, subcutaneous injection of formalin (0.5-5%) produced a characteristic biphasic concentration-related expression of nociceptive behaviours and an increase in paw volume. This study directly demonstrates that formalin can evoke a concentration-dependent local release of adenosine from the rat hind paw. The ability of an adenosine kinase inhibitor and an adenosine deaminase inhibitor to modulate this release is dependent on substrate adenosine concentrations.

The HIV glycoproteins gp41 and gp120 cause rapid excitation in rat cortical slices.

Inflammation and associated excitotoxicity may play important roles in various neurodegenerative diseases including AIDS dementia. Here we show that exposure of rat parietal cortical slices to the HIV glycoproteins gp120 and gp41 triggered very rapid releases of the neurotransmitters glutamate and [(3)H]noradrenaline (NA), and of the neuromodulator adenosine. Gp41 was more effective than gp120 at releasing glutamate and [(3)H]NA, while both glycoproteins were equi-effective at releasing adenosine. The responses to gp120 and gp41 declined rapidly to basal levels following their removal. It seems possible that rapid, inappropriate excitation may occur in the immediate vicinity of HIV infections in the brain, possibly producing some of the transient neurological and psychiatric symptoms associated with AIDS dementia.

Brief communication: cutmarks on a plio-pleistocene hominid from Sterkfontein, South Africa.

Cutmarks inflicted by a stone tool were observed on the right maxilla of Stw 53, an early hominid partial skull from Sterkfontein "Member 5" (South Africa). The morphology of the marks, their anatomical placement, and the lack of random striae on the specimen all support an interpretation of this linear damage as cutmarks. The location of the marks on the lateral aspect of the zygomatic process of the maxilla is consistent with that expected from slicing through the masseter muscle, presumably to remove the mandible from the cranium. Although radioisotopic dates are not available and relative faunal dating of the deposit from which Stw 53 derives is problematic, the morphology of the hominid skull suggests a Plio-Pleistocene age for the specimen. This therefore constitutes the earliest unambiguous evidence that hominids disarticulated the remains of one another.

Jaws and teeth of Australopithecus afarensis from Maka, Middle Awash, Ethiopia.

The Maka locality in Ethiopia's Middle Awash area has yielded new craniodental remains dated to 3.4 million years (myr) in age. These remains are described and assessed functionally and systematically. The fossils are assigned to Australopithecus afarensis. Maka thus joins Hadar and Laetoli as the third major locality yielding this species. As with previous site samples, the Maka collection displays a wide range of size variation. The nearly complete and undistorted MAK-VP-1/12 adult mandible from Maka is an excellent match for Hadar and Laetoli counterparts, confirming the geographic and temporal distribution of A. afarensis. This specimen shows that this taxon is functionally and developmentally hominid in its incisor/canine/premolar complex. A postulated evolutionary trajectory through A. anamensis to A. afarensis would have involved postcanine megadontia and other adaptations to a more heavily masticated diet relative to the earlier Ardipithecus ramidus.

Morphological analysis of the mammalian postcranium: a developmental perspective.

The past two decades have greatly improved our knowledge of vertebrate skeletal morphogenesis. It is now clear that bony morphology lacks individual descriptive specification and instead results from an interplay between positional information assigned during early limb bud deployment and its "execution" by highly conserved cellular response programs of derived connective tissue cells (e.g., chondroblasts and osteoblasts). Selection must therefore act on positional information and its apportionment, rather than on more individuated aspects of presumptive adult morphology. We suggest a trait classification system that can help integrate these findings in both functional and phylogenetic examinations of fossil mammals and provide examples from the human fossil record.

Adenosine: a mediator of interleukin-1beta-induced hippocampal synaptic inhibition.

Interleukin-1 (IL-1) is a pleotrophic cytokine implicated in a variety of central activities, including fever, sleep, ischemic injury, and neuromodulatory responses, such as neuroimmune, and neuroendocrine interactions. Although accumulating evidence is available regarding the expression pattern of this cytokine, its receptors in the CNS, and its mechanistic profile under pathological levels, it is unclear whether this substance modulates central neurons under physiological concentrations. Further, in light of the functional and spatial overlap between the adenosine and IL-1 systems, it is not known whether these two systems are coupled. We report here that, in rat brain slices, brief application of sub-femtomolar IL-1beta causes a profound decrease of glutamate transmission, but not GABAergic inhibition, in hippocampal CA1 pyramidal neurons. This decrease by IL-1beta is prevented by pharmacological blockade of adenosine A1 receptors. In addition, we show that IL-1beta failed to suppress glutamate transmission at room temperature. Because the production and release of adenosine in the CNS is thought to be metabolically dependent, this observation suggests that one of the functions of IL-1beta is to increase the endogenous production of adenosine. Together, these data suggest for the first time that sub-femtomolar levels of IL-1 can effectively modulate glutamate excitation in hippocampal neurons via an adenosine-dependent mechanism.

The bacterial endotoxin lipopolysaccharide causes rapid inappropriate excitation in rat cortex.

There is mounting evidence that inflammation and associated excitotoxicity may play important roles in various neurodegenerative disorders, such as bacterial infections, Alzheimer's disease, AIDS dementia, and multiple sclerosis. The immunogen E. coli lipopolysaccharide (LPS, endotoxin) has been widely used to stimulate immune/inflammatory responses both systemically and in the CNS. Here, we show that exposure of parietal cortical slices from adult rats to LPS triggered very rapid (<2.5 min) and sustained releases of the neurotransmitters glutamate and noradrenaline, and of the neuromodulator adenosine. The responses to LPS declined rapidly following removal of the LPS and exhibited no tachyphylaxis to repeated exposures to LPS. The detoxified form of LPS had no effect. LPS-evoked release of [3H]noradrenaline, but not of glutamate or adenosine, appears to be partly due to the released glutamate acting at ionotropic receptors on the noradrenergic axons present in the cortical slices. LPS appears to release glutamate, which then acts at non-NMDA receptors to remove the voltage-sensitive Mg2+ block of NMDA receptors, thus permitting NMDA receptors to be activated and noradrenaline release to proceed. It seems possible that rapid, inappropriate excitation may occur in the immediate vicinity of gram-negative bacterial infections in the brain. If similar inappropriate excitations are also triggered by those immunogens specifically associated with Alzheimer's disease (beta-amyloid), AIDS dementia (gp120 and gp41), or multiple sclerosis (myelin basic protein), they might explain some of the acute, transient neurological and psychiatric symptoms associated with these disorders.

Co-administration of adenosine kinase and deaminase inhibitors produces supra-additive potentiation of N-methyl-D-aspartate-evoked adenosine formation in cortex.

Activation of glutamate receptors triggers the release of adenosine, which exerts important inhibitory actions in the brain. Evoked adenosine release is potentiated when either adenosine kinase or adenosine deaminase are inhibited. We studied the effects of concurrent inhibition of adenosine kinase and adenosine deaminase on N-methyl-D-aspartate (NMDA)-evoked formation of extracellular adenosine in slices of rat parietal cortex, to determine if combinations of inhibitors of adenosine kinase and adenosine deaminase can produce supra-additive potentiation of this adenosine formation. Combinations of low concentrations of the adenosine kinase inhibitors 5'-amino-5'-deoxyadenosine (0.2 microM) or 5'-iodotubercidin (0.01 microM) with a low concentration of the adenosine deaminase inhibitor 2'-deoxycoformycin (0.2 microM) produced additive potentiations of NMDA-evoked adenosine release from slices of rat parietal cortex. However, combinations of low concentrations of 5'-amino-5'-deoxyadenosine (0.2 microM) or 5'-iodotubercidin (0.01 microM) with a maximal concentration of 2'-deoxycoformycin (200 microM) produced supra-additive potentiation of NMDA-evoked adenosine release. These findings suggest that such combinations of adenosine kinase inhibitors with adenosine deaminase inhibitors may provide useful strategies for developing therapies to treat disorders associated with excessive NMDA receptor activation, such as seizures, ischemic damage and neurodegenerative diseases.

Effect of protein kinase C activation on N-methyl-D-aspartate-evoked release of adenosine and [3H]Norepinephrine from rat cortical slices.

The role of protein kinase C (PKC) in the N-methyl-D-aspartate (NMDA)-evoked release of adenosine (ADO) and [3H]norepinephrine (NE) from slices of rat parietal cortex was studied. In the absence of Mg++, the PKC activator phorbol 12-myristate 13-acetate (1 microM, PMA) did not release either ADO or [3H]NE, but it potentiated the release of ADO evoked by 20 microM NMDA and the release of -3H-NE evoked by 100 microM NMDA. These potentiating effects of PMA on the NMDA-evoked release of ADO and [3H]NE were reversed by the PKC inhibitor GF109,203X (1 microM). GF109,203X by itself had no effect on the NMDA-evoked release of either ADO or [3H]NE. In the presence of Mg++, PMA did not permit the NMDA-evoked release of [3H]NE to occur. These results indicate that PKC does not play an essential role in the NMDA-evoked release of either ADO or NE. However, activation of PKC potentiates the release of ADO and NE evoked by submaximal concentrations of NMDA. Activation of PKC will have the effect of increasing the inhibitory threshold provided by released ADO when only a few NMDA receptors are activated and will promote and accelerate excitatory responses when most of the available NMDA receptors become activated.

A comparison of (+/-)epibatidine with NMDA in releasing [3H]noradrenaline and adenosine from slices of rat hippocampus and parietal cortex.

We compared (+/-)epibatidine with N-methyl-D-aspartate (NMDA) in releasing adenosine and [3H]noradrenaline from slices of rat hippocampus and parietal cortex. (+/-)Epibatidine (0.1 microM) released [3H]noradrenaline but not adenosine from hippocampal slices incubated either with or without extracellular Mg2+. In contrast, NMDA (300 microM) released much more [3H]noradrenaline and also adenosine from hippocampal slices incubated in medium lacking Mg2+. (+/-)Epibatidine released neither adenosine nor [3H]noradrenaline from slices of rat parietal cortex, in contrast to NMDA which released both substances. These findings suggest that those behavioral responses to (+/-)epibatidine that are mediated by noradrenaline may involve the hippocampus but not the parietal cortex. Moreover, it seems unlikely that any of the behavioral effects of (+/-)epibatidine are mediated by adenosine release in either the parietal cortex or the hippocampus.