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PubMed1990

Transcallosal non-pyramidal cell projections from visual cortex in the cat.

Non-pyramidal cells with transcallosal projections were identified in the area 17/18 border region of the cat by retrograde transport of horseradish peroxidase injected into border region of the opposite hemisphere. From several hundred neurons filled with a Golgi-like diaminobenzidine (DAB) reaction product, seven cells were identified by their radially oriented smooth dendrites as possible non-pyramidal cells. Following thin-sectioning and examination with the electron microscope, four of the neurons proved to be layer IV spiny stellate cells with incompletely filled dendritic spines, and two proved to be layer III pyramidal cells with an incompletely labelled apical dendrite and dendritic spines. The remaining neuron was a non-pyramidal cell whose essentially smooth dendrites were covered with synapses, and whose cell body formed both symmetric and asymmetric synapses with presynaptic terminals. To better assess how many non-pyramidal cells might be labelled, thin sections of the area 17/18 border were surveyed using material processed with tetramethylbenzidine (TMB), and another five labelled non-pyramidal cells with transcallosal projections were identified by the needle-like crystals of TMB reaction product they contained. During the study it became evident that both the DAB and TMB reaction products in the lightly labelled neurons tended to be associated with granules that are 0.5 microns or larger in diameter and that had the characteristics of lysosomes. These granules are also visible in the light microscope as dark puncta. The numbers of puncta in profiles of pyramidal and of non-pyramidal cells in layers II/III and IVa of the area 17/18 border region and in the control acallosal region of area 17 were counted and compared. These comparisons revealed that labelled transcallosally projecting non-pyramidal cells may constitute 10-32% of the non-pyramidal cell population at the area 17/18 border region. Similar values were also obtained for pyramidal cells in this region. Consequently, it is concluded that significant numbers of non-pyramidal cells have axons that project through the corpus callosum to the contralateral hemisphere.

Animals↗
PubMed1991

Morphological organization of rat hippocampal slice cultures.

Using various histological methods, we investigated the cellular and morphological organization of rat hippocampal slice cultures. Many of the typical features of the hippocampus were retained in vitro over a long period of time. The principal cell types of the hippocampus and dentate gyrus, the pyramidal cells and granule cells, were well preserved and matured in vitro. Nonpyramidal cells and gamma-aminobutyric-acid (GABA) cells were also present in slice cultures and exhibited a strikingly similar dendritic appearance at the light microscopic level. Moreover, GABA-immunoreactive cell bodies and presynaptic terminals could be identified at the electron microscopic level; they expressed typical symmetric synaptic contacts with cell bodies and dendrites. The course of the intrinsic hippocampal fiber pathways--the mossy fibers, Schaffer collaterals, and alveus--was generally retained in vitro. Additional aberrant fiber projections could be identified. Finally, three types of nonneuronal cells could be distinguished on the basis of immunocytochemical methods.

Animals↗
PubMed1993

Electron microscopic analysis of adrenalectomy-induced hippocampal granule cell degeneration in the rat: apoptosis in the adult central nervous system.

As described in the preceding paper, adrenalectomy triggers hippocampal granule cell degeneration that begins within days after adrenalectomy, continues for months, and is the only apparent cell death anywhere within the brain. At the light microscopic level, granule cell degeneration is characterized by coalescing of nuclear chromatin into numerous spherical bodies. Since the morphology at the light microscopic level resembled the nuclear morphology characteristic of "apoptosis" rather than "necrosis," we undertook this ultrastructural study to determine if adrenalectomy induces the morphological features characteristic of apoptosis. Electron microscopy revealed coalescing of nuclear chromatin, compaction of cytoplasm, and the budding-off of cytoplasmic bodies that were engulfed by glia. Mitochondria, the Golgi apparatus, and rough endoplasmic reticulum appeared relatively normal early in the process of granule cell degeneration when nuclear changes were prominent. Presynaptic terminals innervating degenerating granule cells appeared normal. Electron-dense degeneration of granule cell axon terminals in association with normal postsynaptic elements of CA3 pyramidal cells highlighted the extraordinary selectivity of adrenalectomy-induced granule cell death. Ten weeks after adrenalectomy, astrocytes were filled with abnormally abundant glial fibrils and neuronal debris. This "apoptotic" morphology produced by adrenalectomy was clearly distinct from the "necrotic" granule cell morphology produced by intrahippocampal injection of the neurotoxin volkensin. These results indicate that, in a manner possibly analogous to castration-induced prostate cell death, loss of adrenal hormone triggers a process in dentate granule cells that causes the morphological changes characteristic of "apoptosis." Thus, adrenal steroids may be obligatory growth factors for dentate granule cells and their loss may initiate a selective process in the mature brain that is unique or that may normally occur only in the developing brain.

Adrenalectomy↗
PubMed1994

Histological and ultrastructural localization of the kainate receptor subunits, KA2 and GluR6/7, in the rat nervous system using selective antipeptide antibodies.

Kainate receptors are found throughout many regions of the brain and presumably contribute to responses of neurons to glutamate and other excitatory amino acids. Two affinity-purified polyclonal antibodies that recognize the kainate binding subunits, KA2 and GluR6, were made using C-terminus peptides. A previous study demonstrated that each antibody is specific for its subunit, although antibody to GluR6 recognizes GluR7 to some extent (hence the designation GluR6/7). Vibratome sections immunostained with either antibody showed light to moderate staining in many structures in the brain as well as in cervical spinal cord, dorsal root and vestibular ganglia, and pineal and pituitary glands. Moderate levels were seen in the olfactory bulb, cerebral cortex, caudate/putamen, and hypothalamus, whereas much of the thalamus was stained lightly. In the hippocampus, CA3 pyramidal cells were stained more densely than CA1 pyramidal cells--the difference more evident with antibody to GluR6/7. In addition, neuropilar staining was densest in the stratum lucidum of the CA3 region. In the brainstem, staining was moderate to moderately dense in a number of sensory, motor, and reticular nuclei. The moderately dense staining in the reticulothalamic nucleus and pontine nuclei with antibody to GluR6/7 may represent its recognition of GluR7. In the cerebellum, staining was moderate in granular and molecular layers with antibody to KA2 and in the molecular layer with antibody to GluR6/7, whereas it was moderately dense to dense in the granular layer with the GluR6/7 antibody. Outside of the brain, densest staining was seen with antibody to KA2 in the intermediate lobe of the pituitary gland. Ultrastructural localization of immunostaining was examined in the hippocampus, cerebral cortex, and cerebellar cortex. Typically, major staining was in postsynaptic densities apposed by unstained presynaptic terminals with round or mainly round vesicles and in associated dendrites. The light microscope pattern of staining was fairly similar to that of previous [3H]kainate binding and in situ hybridization studies. In addition, comparison with previous studies on distribution of other types of glutamate receptors indicates that KA2 and GluR6/7 are found with various other subunits in many of the same cell populations throughout the nervous system.

Amino Acid Sequence↗
PubMed2006

Characterization of a mouse strain expressing Cre recombinase from the 3' untranslated region of the dopamine transporter locus.

Dopamine (DA) neurotransmission has been implicated in several neurological and psychiatric disorders. The dopamine transporter (DAT) is highly expressed in dopaminergic neurons of the ventral mesencephalon and regulates neurotransmission by transporting DA back into the presynaptic terminals. To mediate restricted DNA recombination events into DA neurons using the Cre/loxP technology, we have generated a knockin mouse expressing Cre recombinase under the transcriptional control of the endogenous DAT promoter. To minimize interference with DAT function by preservation of both DAT alleles, Cre recombinase expression was driven from the 3' untranslated region (3'UTR) of the endogenous DAT gene by means of an internal ribosomal entry sequence. Crossing this murine line with a LacZ reporter showed colocalization of DAT immunocytochemistry and beta-galactosidase staining in all regions analyzed. This knockin mouse can be used for generating tissue specific knockouts in mice carrying genes flanked by loxP sites, and will facilitate the analysis of gene function in dopaminergic neurons.

3' Untranslated Regions↗
PubMed1995

Population genetic study of the human dopamine transporter gene (DAT1).

The human dopamine transporter gene, DAT1, acts to transport released dopamine into presynaptic terminals of the brain. The possibility that the DAT1 gene plays a role in genetic diseases of the brain has led to studies of DAT1 in several psychiatric and neurological disorders. Previous sequence analysis of DAT1 revealed a 40-bp repeat in the 3' end of the gene. In order to identify all potential alleles for this VNTR marker a population database was established. One thousand seventy-four unrelated individuals were screened by PCR for the region containing the 40 bp repeat. Allele frequency differences were found between black Americans and Caucasians or Hispanics but no differences were observed between Caucasians and Hispanics. A previously unreported allele was detected in all three populations. Thus, we have shown that screening a large population identifies new alleles and generates more accurate allele frequencies.

Alleles↗
PubMed2004

Potassium attenuates zinc-induced death of cultured cortical astrocytes.

Transient global ischemia induces CA1 hippocampal neuronal death without astrocyte death, perhaps mediated in part by the toxic translocation of zinc from presynaptic terminals to postsynaptic neurons. We tested the hypothesis that cellular depolarization, which occurs in the ischemic brain due to increased extracellular potassium and energy failure, might contribute to astrocyte resistance to zinc-induced death. We previously reported that neurons in mixed cortical neuronal-astrocyte cultures were more vulnerable to a 5-15-min exposure to Zn(2+) than astrocytes in the same cultures. In the present report, we show that (1) neurons in isolation or in conjunction with astrocytes were 2-3-fold more sensitive to a 15-min nondepolarizing Zn(2+) exposure than are glia; (2) KCl-induced depolarization attenuated glial vulnerability to zinc toxicity but potentiated neuronal vulnerability to zinc toxicity; (3) Zn(2+)-induced glial death was attenuated by T-type Ca(2+) channel blockade, as well as compounds that increase NAD(+) levels; and (4) both astrocytic (65)Zn(2+) accumulation and the increase in astrocytic [Zn(2+)](i) induced by Zn(2+) exposure were also attenuated by depolarization or T-type Ca(2+) channel blockers. Zn(2+)-induced cell death in astrocytes was at least in part apoptotic, as caspase-3 was activated, and the caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone partially attenuated Zn(2+)-induced death. The levels of peak [Zn(2+)](i) achieved in astrocytes during this toxic nondepolarizing Zn(2+) exposure (250 nM) were substantially greater than those achieved in neurons (40 nM). In glia, exposure to 400 microM Zn(2+) induced a 13-mV depolarization, which can activate T-type Ca(2+) channels. This Zn(2+)-induced astrocyte death, like neuronal death, was attenuated by the addition of pyruvate or niacinamide to the exposure medium.

Animals↗
PubMed1993

Quantal and subquantal GABAergic transmissions in cultured rat hippocampal neurons.

At the neuromuscular junction, spontaneous miniature excitatory synaptic currents mediated by acetylcholine are considered elementary, "quantal" transmissions. These miniature conductances can be quantitatively dichotomized into a large-mode class whose mode is the mean of a normal, bell-shaped distribution and a small-mode class whose distribution is skewed to lower values with its mode being a fraction of the large-mode class. The large-mode class constitutes the population of synaptic signals originally utilized to formulate tenets of "quantal" transmission, which have been tacitly adopted in more recent studies of fast transmission at central synapses. Large- and small-mode conductance classes of inhibitory synaptic elementary conductances mediated by GABA have now been recorded in cultured hippocampal neurons (Vautrin J, Schaffner AE, Barker JL, 1991, Neurosci Lett 138:67). Pairs of hippocampal neurons were patch-recorded at optimal signal-to-noise and, using time course analysis, two elementary fluctuations (0.1-0.3 nS and 1-2 nS) were found within synaptic conductances evoked either by presynaptic action potentials or by presynaptic terminal stimulation. These results were interpreted with a simple model that shows how different frequencies of unitary GABA release can generate either small-mode, skew-distributed conductance (0.5-3 kHz) or large-mode, normally-distributed conductances (> or = 10 kHz). Only the latter satisfies the original tenets of the classic quantal theory.

Action Potentials↗
PubMed1988

Morphology of synapses in the autonomic nervous system.

The ultrastructure of synapses in the autonomic nervous system is reviewed. The synaptic organization of the parasympathetic ganglia is relatively simple. Preganglionic axons form synapses either on the soma or on short perikaryal processes of the ganglionic neurons. The presynaptic terminals have a cholinergic morphology and contain mainly small clear vesicles with a few large dense cored vesicles. A few neuropeptides have been localized to the large dense cored vesicles of these terminals. The postganglionic parasympathetic axons ramify within their target tissues where they form close associations, but not true synaptic contacts. Sites of release of transmitter are recognized morphologically as varicosities along the length of the axon that contain clusters of small clear vesicles with a few large dense cored vesicles. The organization of the sympathetic nervous system is somewhat more complex. In addition to acetylcholine, enkephalin also exists in these terminals, probably in the large dense cored vesicles. There are at least three types of ganglion cell neurons in the paravertebral portion of the sympathetic nervous system: those that contain norepinephrine alone, those that contain norepinephrine along with neuropeptide Y, and those that contain acetylcholine and vasoactive intestinal polypeptide. The first type provides innervation to the parenchyma of the target tissues, while the second mainly innervates blood vessels. The third type innervates the sweat glands. In the prevertebral ganglia, a fourth type of neuron exists that contains norepinephrine and somatostatin. This neuron probably innervates the gut. Preganglionic terminals of the cholinergic type form synaptic connections mainly with the dendrites of the sympathetic ganglion neurons. In addition to the types of synapses described for the paravertebral ganglia, neurons in the prevertebral ganglia receive synaptic connections from dorsal root ganglia and from the enteric nervous system. The sympathetic ganglia also contain interneurons that receive preganglionic synapses and form efferent synapses with some of the principal ganglion cells. The interneurons have been shown to contain a variety of transmitters, including norepinephrine, epinephrine, dopamine, serotonin, and a number of neuropeptides. The postganglionic sympathetic axons have a similar morphology to the parasympathetic axons. They form networks in their targets, and the axons display varicosities with concentrations of both small and large vesicles. After appropriate fixation, these vesicles are seen to possess dense cores.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2001

Microglia in neuroregeneration.

Microglia has the potential to produce and release a range of factors that directly and/or indirectly promote regeneration in the injured nervous system. The overwhelming evidence indicates, however, that this potential is generally not expressed in vivo. Activated microglia may enhance neuronal degeneration following axotomy, thereby counteracting functional recovery. Microglia does not seem to contribute significantly to axonal outgrowth after peripheral nerve injury, since this process proceeds uneventful even if perineuronal microglia is eliminated. The phagocytic phenotype of microglia is highly suppressed during Wallerian degeneration in the central nervous system. Therefore, microglia is incapable of rapid and efficient removal of myelin debris and its putative growth inhibitory components. In this way, microglia may contribute to regeneration failure in the central nervous system. Structural and temporal correlations are compatible with participation by perineuronal microglia in axotomy-induced shedding of presynaptic terminals, but direct evidence for such participation is lacking. Currently, the most promising case for a promoting effect on neural repair by activated microglia appears to be as a mediator of collateral sprouting, at least in certain brain areas. However, final proof for a critical role of microglia in these instances is still lacking. Results from in vitro studies demonstrate that microglia can develop a regeneration supportive phenotype. Altering the microglial involvement following neural injury from a typically passive or even counterproductive state and into a condition where these cells are actively supporting regeneration and plasticity is, therefore, an exciting challenge and probably a realistic goal.

Central Nervous System↗
PubMed2004

Technique to quantify local clustering of synaptic vesicles using single section data.

Synaptic vesicles are organelles that specialize in the storage of a neurotransmitter that continuously undergo an exo-endocytotic cycle. During this cycle vesicles change their positions within a presynaptic terminal and their numbers as well as spatial arrangement can provide insight into a neurotransmitter turnover. This article introduces a technique based on the nearest-neighbor formalism to quantify the proximity of vesicles to active zones and vesicle clustering in different regions of a terminal. The technique, implemented in a software package, uses the two-dimensional coordinates of features identified in digitized electron micrographs as an input. It has been validated in the analysis of asymmetric synapses of the rat hippocampal CA1 stratum radiatum affected by transient cerebral ischemia. It was shown that a 15-minute-long ischemic episode influenced the spatial arrangement of vesicles that were more distant from active zones and had larger intervesicle spacings with respect to the control. The latter effect was apparently stronger within 200 nm distance of active zones.

Animals↗
PubMed2001

Glial cell death induced by overexpression of alpha-synuclein.

alpha-Synuclein is present in intracellular protein aggregates that are hallmarks of common neurodegenerative disorders including Parkinson disease, dementia with Lewy bodies, and multiple system atrophy. alpha-Synuclein is localized in neurons and presynaptic terminals. Under pathological conditions, however, it is also found in glia. The role of alpha-synuclein in glial cells and its relevance to the molecular pathology of neurodegenerative diseases is presently unclear. To investigate the consequence of alpha-synuclein overexpression in glia, we transfected U373 astrocytoma cells with vectors encoding wild-type human alpha-synuclein or C-terminally truncated synuclein fused to red fluorescent protein. alpha-synuclein immunocytochemistry of transfected astroglial cells revealed diffuse cytoplasmic labeling associated with discrete inclusions both within cell bodies and processes. Susceptibility to oxidative stress was increased in astroglial cells overexpressing alpha-synuclein, particularly in the presence of cytoplasmic inclusions. Furthermore, overexpression of alpha-synuclein induced apoptotic death of astroglial cells as shown by TUNEL staining. Our in vitro model is the first to replicate salient features of the glial pathology associated with alpha-synucleinopathies. It provides a simple testbed to further explore the cascade of events that leads to apoptotic glial cell death in some of these disorders; it may also be useful to assess the effects of therapeutic interventions including antioxidative and antiapoptotic strategies.

Apoptosis↗
PubMed2005

Heterogeneous expression and regulation of hippocampal prostaglandin E2 receptors.

Although prostaglandin E2 (PGE2) has been shown to be critical to hippocampal synaptic signaling and neuronal survival, it is still not clear which subtypes of PGE2 receptors (EPs) are expressed and how these EPs are regulated in the hippocampus. To address these questions, the expression of the EPs was profiled in the hippocampus. Messenger RNAs and proteins of the four receptors, EP 1-4, were detected both in the hippocampus and in the neocortex. EP 2 and EP 3 appeared in greater abundance, whereas EP 1 and EP 4 were barely detectable. EP 1, EP 2 and EP 4 were mainly colocalized with synaptophysin, suggesting the presence of EP 1, EP 2, and EP 4 in presynaptic terminals. It appeared that interleukin-1 beta increased the expression of EP 2 and EP 4 mRNAs. A blockade of synaptic transmission with either tetrodotoxin or MK-801 plus 6,7-dinitroquinoxaline-2,3-dione (DNQX) for 6 hr increased EP 3 and EP 4 mRNA, whereas high K(+) (90 mM) or 4-aminopyridine enhanced EP 2 and EP 4. The EP 1 level did not change significantly under these conditions. The expressions of EP 2, EP 4, and EP 3 were further elevated or reduced in neurons treated with high K(+) for 24 hr. However, mRNA of EP 3 was down-regulated in neurons treated with tetrodotoxin or MK-801 plus DNQX for 24 hr. In addition, both EP 2 and EP 4 mRNAs were up-regulated within 4 hr after high-frequency stimulation associated with long-term potentiation induction in hippocampal slices. Our results indicate that the four EPs are heterogeneously expressed in the hippocampus, and their expression is differentially regulated by neuronal activities, suggesting that EPs may actively participate in hippocampal synaptic transmission and plasticity.

Animals↗
PubMed2005

alpha1-Adrenergic modulation of synaptic input to Purkinje neurons in rat cerebellar brain slices.

The inhibitory activity in the cerebellar network, as investigated in acute brain slices from 14-20 days old rats, is modulated by alpha1-adrenergic stimulation. The specific alpha1-adrenoceptor agonist phenylephrine (PhE; 10 microM) or the alpha-adrenoceptor agonist 6-fluoronoradrenaline (10 microM) increases the frequency and the amplitude of spontaneous postsynaptic currents (sPSC) in Purkinje neurons. The effects are sensitive to the alpha1-adrenoceptor antagonists prazosin (30 microM) and phentolamine (10 microM). The PhE-induced augmentation is suppressed when phospholipase C is blocked by preincubation with U73122 (10 microM) but is not affected by inhibition of protein kinases with H7 (10 microM) or GF109203X (10 microM). Involvement of intracellular Ca(2+) stores was shown by a reduced PhE effect after blocking of SERCA pumps with cyclopiazonic acid (30 microM) and thapsigargin (1 microM). The persistence of the PhE effect on the frequency of miniature postsynaptic currents, as recorded in presence of tetrodotoxin, indicates a presynaptic localization of the alpha1-adrenoceptors. A block of voltage-gated Ca(2+) channels with nifedipine, verapamil, or omega-conotoxin MVIIC did not suppress the PhE-induced increase of the frequency and amplitude of sPSC. The results suggest that alpha1-adrenoceptors at presynaptic terminals mediate an increase of the spontaneous synaptic inhibition of Purkinje neurons in the cerebellar cortex via release of Ca(2+) from intracellular stores.

2-Amino-5-phosphonovalerate↗
PubMed2006

Heavy chain of cytoplasmic dynein is a major component of the postsynaptic density fraction.

A protein with an apparent molecular size of 490 kDa was found in the postsynaptic density (PSD) fraction isolated from porcine cerebral cortices and rat forebrains, and this 490 kDa protein accounted for approximately 3% of the total protein of these samples. Matrix-assisted laser desorption ionization-time of flight mass spectrometric and Western blotting analyses consistently indicated that this 490 kDa protein consisted primarily of the heavy chain of cytoplasmic dynein (cDHC). Immunocytochemical analyses showed that cDHC was found in 92% and 89% of the phalloidin-positive protrusions that were themselves associated with discrete clusters of synaptophysin, a presynaptic terminal marker, and PSD-95, a postsynaptic marker, on neuronal processes, respectively. Quantitative Western blotting analyses of various subcellular fractions isolated from porcine cerebral cortices and rat forebrains further showed that not only the heavy but also the intermediate chains of dynein are enriched in the PSD fraction. Cytoplasmic dynein is a microtubule-associated motor protein complex that drives the movement of various cargos toward the minus ends of microtubules and plays many other diverse functions in the cell. Our results that cDHC is a major component of the PSD fraction, that both dynein heavy and intermediate chains are enriched in the PSD fraction and that cDHC is present in dendritic spines raise the possibilities that cytoplasmic dynein may play structural and functional roles in the postsynaptic terminal.

Animals↗
PubMed1977

Endotoxin alters spontaneous transmitter release at the frog neuromuscular junction.

The direct neurotoxic effects of E. coli endotoxin (ETX) on spontaneous transmitter release were tested at the frog sartorius muscle neuromuscular junction. Spontaneous transmitter release was monitored by intracellularly recording miniature end-plate potentials (MEPPs). Junctions were continuously exposed to standard concentrations of 10 microgram/ml of 3 ETX samples, 2 of which produced a significant elevation of MEPP frequency followed by a decline of frequency to very low rates. The third ETX sample, known to have a decreased canine lethality, was without effect on MEPP frequency. No significant changes in MEPP amplitude were evident. The rate of change in MEPP frequency, but not the peak frequency, was reduced by lowering ETX concentrations. Alterations in MEPP frequency induced by ETX were prevented by removing Ca++ and antagonized by high [K+]out. The results suggest that ETX alters ion conductance channels, particularly those for Ca++, in the presynaptic terminal membrane.

Action Potentials↗
PubMed1981

Endogenous activation of guanylate cyclase in synaptosomal soluble fraction from rat brain.

Guanylate cyclase in crude mitochrondrial (P2) soluble fraction prepared from rat brain, obtained by a hypo-osmotic treatment of P2, showed extremely higher activity than that in the same fraction from other organs. In addition the soluble fraction obtained from synaptosomes (P2 - B) contained the highest enzyme activity among other subfractions of the cerebral P2 examined. Guanylate cyclase activity in the synaptosomal soluble fraction was, however, markedly suppressed by various compounds reacting with free radicals. These results suggest that guanylate cyclase in the synaptosomal soluble fraction may be activated endogenously be a free radical and involved in the regulatory mechanisms for cyclic guanosine monophosphate (cyclic GMP) at presynaptic terminals.

Animals↗
PubMed1989

Axonal transport of a clathrin uncoating ATPase (HSC70): a role for HSC70 in the modulation of coated vesicle assembly in vivo.

Clathrin plays an important role in many cellular processes, including endocytosis, secretion, and sorting of membranous organelles. Both the neuronal cell body and presynaptic terminals contain numerous coated vesicles, but few are detectable in the axonal regions that connect these two regions of the neuron. Clathrin heavy chains, light chains, and assembly proteins have all been shown to be axonally transported as part of slow component b (SCb). However, the paucity of coated vesicles present in the axon indicates the existence of a mechanism regulating clathrin coated assembly in vivo. A clathrin uncoating ATPase has been described that binds in stoichiometric amounts to clathrin and dissociates clathrin coats from vesicles in the presence of ATP in vitro. This clathrin uncoating ATPase is a major cytosolic protein of Mr 70 kD in bovine brain, forming 1% of soluble brain protein, and appears to be homologous with a constitutively expressed 70 kd heat shock protein (HSC70). We report here that a major 70 kD protein present in the SCb rate component of axonal transport is identical with HSC70 by both biochemical and immunochemical criteria. The cotransport of HSC70 with clathrin in SCb of axonal transport is consistent with a role for HSC70 in vivo in the regulation of clathrin function during axonal transport.

Adenosine Triphosphatases↗