Martin Bohlen - Profile on Academia.edu (original) (raw)
Papers by Martin Bohlen
Database for the design of tailored light delivery for optogenetics in non-human primates
This study analyzes the light emission pattern of tapered optical fibers (TFs) across different n... more This study analyzes the light emission pattern of tapered optical fibers (TFs) across different numerical apertures (NA), core diameters, laser input powers, cone angles, and light injection angles. The results will be used to tailor optical fibers for optimal light delivery to photosensitive neuronal actuators in the non-human primate (NHP) brain.
Superior colliculus projections to target populations in the supraoculomotor area of the macaque monkey
Visual Neuroscience, 2021
A projection by the superior colliculus to the supraoculomotor area (SOA) located dorsal to the o... more A projection by the superior colliculus to the supraoculomotor area (SOA) located dorsal to the oculomotor complex was first described in 1978. This projection’s targets have yet to be identified, although the initial study suggested that vertical gaze motoneuron dendrites might receive this input. Defining the tectal targets is complicated by the fact the SOA contains a number of different cell populations. In the present study, we used anterograde tracers to characterize collicular axonal arbors and retrograde tracers to label prospective SOA target populations in macaque monkeys. Close associations were not found with either superior or medial rectus motoneurons whose axons supply singly innervated muscle fibers. S-group motoneurons, which supply superior rectus multiply innervated muscle fibers, appeared to receive a very minor input, but C-group motoneurons, which supply medial rectus multiply innervated muscle fibers, received no input. A number of labeled boutons were observe...
Cerebellar projections to the macaque midbrain tegmentum: Possible near response connections
Visual Neuroscience, 2021
Since most gaze shifts are to targets that lie at a different distance from the viewer than the c... more Since most gaze shifts are to targets that lie at a different distance from the viewer than the current target, gaze changes commonly require a change in the angle between the eyes. As part of this response, lens curvature must also be adjusted with respect to target distance by the ciliary muscle. It has been suggested that projections by the cerebellar fastigial and posterior interposed nuclei to the supraoculomotor area (SOA), which lies immediately dorsal to the oculomotor nucleus and contains near response neurons, support this behavior. However, the SOA also contains motoneurons that supply multiply innervated muscle fibers (MIFs) and the dendrites of levator palpebrae superioris motoneurons. To better determine the targets of the fastigial nucleus in the SOA, we placed an anterograde tracer into this cerebellar nucleus in Macaca fascicularis monkeys and a retrograde tracer into their contralateral medial rectus, superior rectus, and levator palpebrae muscles. We only observed...
Adeno-Associated Virus Capsid-Promoter Interactions in the Brain Translate from Rat to the Nonhuman Primate
Human Gene Therapy, 2020
Recently, we established an AAV9 capsid-promoter interaction that directly determined cell-specif... more Recently, we established an AAV9 capsid-promoter interaction that directly determined cell-specific gene expression across two synthetic promoters, Cbh and CBA, in the rat striatum. The present studies not only expand this capsid-promoter interaction to include another promoter in the rat striatum, but also establishes AAV capsid-promoter interactions in the non-human primate brain. When AAV9 vectors were injected into the rat striatum, the minimal synthetic promoter JetI drove GFP gene expression predominantly in oligodendrocytes. However, similar to our previous findings, the insertion of 6 alanines into VP1/VP2 of the AAV9 capsid (AAV9AU) significantly shifted JetI-driven GFP gene expression to neurons. In addition, previous retrograde tracing studies in the non-human primate brain also revealed the existence of a capsid-promoter interaction. When rAAV2-Retro vectors were infused into the frontal eye field (FEF) of rhesus macaques, local gene expression was prominent using either the hybrid chicken beta actin (CAG) or human synapsin (hSyn) promoters. However, only the CAG promoter, not the hSyn promoter, led to gene expression in the ipsilateral claustrum and contralateral FEF. Conversely, infusion of rAAV2-retro-hSyn vectors, but not rAAV2-retro-CAG, into the macaque superior colliculus led to differential and selective retrograde gene expression in cerebellotectal afferent cells. Clearly, this differential promoter/capsid expression profile could not be attributed to promoter inactivation from retrograde transport of the rAAV2-Retro vector. In summary, we document the potential for AAV capsid/promoter interactions to impact cell specific gene expression across species, experimental manipulations and engineered capsids, independent of capsid permissivity.
Neuron, 2020
Highlights d Openly shared, non-human primate optogenetics data resource d Over 500 entries of un... more Highlights d Openly shared, non-human primate optogenetics data resource d Over 500 entries of unpublished data d Efficiency analysis of experimental parameters d Advice from leading NHP optogenetics labs
Frontiers in Neuroanatomy, 2019
Reliable viral vector-mediated transgene expression in primate motoneurons would improve our abil... more Reliable viral vector-mediated transgene expression in primate motoneurons would improve our ability to anatomically and physiologically interrogate motor systems. We therefore investigated the efficacy of replication defective, early region 1-deleted canine adenovirus type-2 (CAV-2) vectors for mediating transgene expression of fluorescent proteins into brainstem motoneurons following craniofacial intramuscular injections in four rhesus monkeys (Macaca mulatta). Vector injections were placed into surgically identified and isolated craniofacial muscles. After a 1-to 2-month survival time, animals were sacrificed and transgene expression was assessed with immunohistochemistry in the corresponding motoneuronal populations. We found that injections of CAV-2 into individual craniofacial muscles at doses in the range of ∼10 10 to 10 11 physical particles/muscle resulted in robust motoneuronal transduction and expression of immunohistochemically identified fluorescent proteins across multiple animals. By using different titers in separate muscles, with the resulting transduction patterns tracked via fluorophore expression and labeled motoneuron location, we established qualitative dose-response relationships in two animals. In one animal that received an atypically high titer (5.7 × 10 11 total CAV-2 physical particles) distributed across numerous injection sites, no transduction was detected, likely due to a retaliatory immune response. We conclude that CAV-2 vectors show promise for genetic modification of primate motoneurons following craniofacial intramuscular injections. Our findings warrant focused attention toward the use of CAV-2 vectors to deliver opsins, DREADDs, and other molecular probes to improve genetics-based methods for primate research. Further work is required to optimize CAV-2 transduction parameters. CAV-2 vectors encoding proteins could provide a new, reliable route for modifying activity in targeted neuronal populations of the primate central nervous system.
Journal of Comparative Psychology, 2019
Decisions often involve the consideration of multiple cues, each of which may inform selection on... more Decisions often involve the consideration of multiple cues, each of which may inform selection on the basis of learned probabilities. Our ability to use probabilistic inference for decisions is bounded by uncertainty and constraints such as time pressure. Previous work showed that when humans choose between visual objects in a multiple-cue, probabilistic task, they cope with time pressure by discounting the least informative cues, an example of satisficing or "good enough" decision-making. We tested two rhesus macaques (Macaca mulatta) on a similar task to assess their capacity for probabilistic inference and satisficing in comparison with humans. On each trial, a monkey viewed two compound stimuli consisting of four cue dimensions. Each dimension (e.g., color) had two possible states (e.g., red or blue) with different probabilistic weights. Selecting the stimulus with highest total weight yielded higher odds of receiving reward. Both monkeys learned the assigned weights at high accuracy. Under time pressure, both monkeys were less accurate as a result of decreased use of cue information. One monkey adopted the same satisficing strategy used by humans, ignoring the least informative cue dimension. Both monkeys, however, exhibited a strategy not reported for humans, a "group-the-best" strategy in which the top two cues were used similarly despite their different assigned weights. The results validate macaques as an animal model of probabilistic decision-making, establishing their capacity to discriminate between objects using at least four visual dimensions simultaneously. The time pressure data suggest caution, however, in using macaques as models of human satisficing.
The Anatomical Record, 2019
The organization of extraocular muscles (EOMs) and their motor nuclei was investigated in the mou... more The organization of extraocular muscles (EOMs) and their motor nuclei was investigated in the mouse due to the increased importance of this model for oculomotor research. Mice showed a standard EOM organization pattern, although their eyes are set at the side of the head. They do have more prominent oblique muscles, whose insertion points differ from those of frontal-eyed species. Retrograde tracers revealed that the motoneuron layout aligns with the general vertebrate plan with respect to nuclei and laterality. The mouse departed in some significant respects from previously studied species. First, more overlap between the distributions of muscle-specific motoneuronal pools was present in the oculomotor nucleus (III). Furthermore, motoneuron dendrites for each pool filled the entire III and extended beyond the edge of the abducens nucleus (VI). This suggests mouse extraocular motoneuron afferents must target specific pools based on features other than dendritic distribution and nuclear borders. Second, abducens internuclear neurons are located outside the VI. We concluded this because no unlabeled abducens internuclear neurons were observed following lateral rectus muscle injections and because retrograde tracer injections into the III labeled cells immediately ventral and ventrolateral to the VI, not within it. This may provide an anatomical substrate for differential input to motoneurons and internuclear neurons that This article includes AR WOW Video. Video can be viewed at https://players.brightcove.net/656326989001/default_default/ index.html?videoId=6032629801001 Abbreviations: 4V = fourth ventricle; aVI = accessory abducens nucleus; BC = brachium conjunctivum; CA = cerebral aqueduct; CG = central gray; DR = dorsal raphe; EWcp = centrally projecting Edinger-Westphal nucleus; III = oculomotor nucleus; IO = inferior oblique muscle; IV = trochlear nucleus; LP = levator palpebrae superioris muscle; LR = lateral rectus muscle; MLF = medial longitudinal fasciculus; MR = medial rectus muscle; MRF = mesencephalic reticular formation; ON = optic nerve;
Molecular and Cellular Endocrinology, 2017
Three forms of familial primary aldosteronism have been recognized. Familial Hyperaldosteronism t... more Three forms of familial primary aldosteronism have been recognized. Familial Hyperaldosteronism type 1 (FH1) or dexamethasone suppressible hyperaldosteronism, FH2, the most common form of as yet unknown cause(s), and FH3. FH3 is due to activating mutations of the potassium channel gene KCNJ5 that increase constitutive and angiotensin II-induced aldosterone synthesis. In this study we examined the cellular distribution of CYP11B2, CYP11B1, CYP17A1 and KCNJ5 in adrenals from two FH3 siblings using immunohistochemistry and immunofluorescence and obtained unexpected results. The adrenals were markedly enlarged with loss of zonation. CYP11B2 was expressed sporadically throughout the adrenal cortex. CYP11B2 was most often expressed by itself, relatively frequently with CYP17A1, and less frequently with CYP11B1. KCNJ5 was co-expressed with CYP11B2 and in some cells with CYP11B1. This aberrant co-expression of enzymes likely explains the abnormally high secretion rate of the hybrid steroid, 18-oxocortisol.
The Journal of comparative neurology, Jan 2, 2016
In this study, two points related to the pattern of innervation of the extraocular muscles were e... more In this study, two points related to the pattern of innervation of the extraocular muscles were explored. 1. Species differences exist in the location of the motoneurons supplying multiply innervated fibers (MIFs) and singly innervated fibers (SIFs) in eye muscles. MIF motoneurons are located outside the extraocular nuclei in primates, but are intermixed with SIF motoneurons within rat extraocular nuclei. To test whether this difference is related to visual capacity and frontal placement of eyes, we injected retrograde tracers into the medial rectus muscle of the cat, a highly visual, non-primate with frontally placed eyes. Distal injections labeled smaller MIF motoneurons located ventrolaterally and rostrally within the oculomotor nucleus (III). More central injections also labeled a separate population of larger cells, located dorsally in III. Thus, the cat shares the feature of having MIF motoneurons located within the bounds of III with the nocturnal rat. On the other hand, like...
Journal of AAPOS : the official publication of the American Association for Pediatric Ophthalmology and Strabismus / American Association for Pediatric Ophthalmology and Strabismus, 2016
Extraocular proprioception has been shown to participate in spatial perception and binocular alig... more Extraocular proprioception has been shown to participate in spatial perception and binocular alignment. Yet the physiological approaches used to study this sensory signal are limited because proprioceptive signaling takes place at the same time as visuomotor signaling. It is critical to dissociate this sensory signal from other visuomotor events that accompany eye movements. We present a novel noninvasive and quantifiable method for probing extraocular proprioception independent of other visuomotor processing by attaching a rare-earth magnet to a real-time model eye and placing an electromagnet <20 mm from the eye. An electromagnet can increase or decrease angular displacements and velocities of the model eye. Electromagnetic activation rapidly affected (<2 ms) the rotation kinematics of the eye, which were correlated linearly with both the current supply and the distance of the electromagnet relative to the eye. This method circumvented the constraints of conventional physiol...
Physiology & behavior, Jan 11, 2015
Exposure to unpredictable chronic mild stress (CUS) is a commonly used protocol in rats that is r... more Exposure to unpredictable chronic mild stress (CUS) is a commonly used protocol in rats that is reported to evoke antidepressant-reversible behaviors such as loss of preference for sweetened water solution which is taken as an analog of the anhedonia seen in major depression. However, the induction of anhedonic-like behavior by chronic mild stress, gauged by an animal's preference for sucrose solution, is not fully reproducible and consistent across laboratories. In this study, we compared a widely used behavioral marker of anhedonia - the sucrose preference test, with another phenotypic marker of emotional valence, social interaction-associated ultrasonic vocalizations as well as a marker of an anxiety-like phenotype, novelty-suppressed feeding, and cognitive performance in the eight arm radial maze task in adult male Sprague-Dawley rats. Chronic four-week exposure to unpredictable mild stressors resulted in 1) attenuation of social interaction-associated ultrasonic vocalizatio...
Using rAAV2-retro in rhesus macaques: Promise and caveats for circuit manipulation
Journal of Neuroscience Methods, 2020
BACKGROUND Recent genetic technologies such as opto- and chemogenetics allow for the manipulation... more BACKGROUND Recent genetic technologies such as opto- and chemogenetics allow for the manipulation of brain circuits with unprecedented precision. Most studies employing these techniques have been undertaken in rodents, but a more human-homologous model for studying the brain is the nonhuman primate (NHP). Optimizing viral delivery of transgenes encoding actuator proteins could revolutionize the way we study neuronal circuits in NHPs. NEW METHOD: rAAV2-retro, a popular new capsid variant, produces robust retrograde labeling in rodents. Whether rAAV2-retro's highly efficient retrograde transport would translate to NHPs was unknown. Here, we characterized the anatomical distribution of labeling following injections of rAAV2-retro encoding opsins or DREADDs in the cortico-basal ganglia and oculomotor circuits of rhesus macaques. RESULTS rAAV2-retro injections in striatum, frontal eye field, and superior colliculus produced local labeling at injection sites and robust retrograde labeling in many afferent regions. In every case, however, a few brain regions with well-established projections to the injected structure lacked retrogradely labeled cells. We also observed robust terminal field labeling in downstream structures. COMPARISON WITH EXISTING METHOD(S) Patterns of labeling were similar to those obtained with traditional tract-tracers, except for some afferent labeling that was noticeably absent. CONCLUSIONS rAAV2-retro promises to be useful for circuit manipulation via retrograde transduction in NHPs, but caveats were revealed by our findings. Some afferently connected regions lacked retrogradely labeled cells, showed robust axon terminal labeling, or both. This highlights the importance of anatomically characterizing rAAV2-retro's expression in target circuits in NHPs before moving to manipulation studies.
Behav Brain Res., 2014
h i g h l i g h t s
Journal of Neuroscience Methods, 2009
The latency of mice and rats to fall from the accelerating rotarod can differ markedly between la... more The latency of mice and rats to fall from the accelerating rotarod can differ markedly between laboratories using the same brand of rod as well as between studies using different kinds of rods. These discrepancies can arise from different rod diameters, surface textures, test protocols, or laboratory environmental factors beyond the test itself, but it is also possible that the actual acceleration rates of the different rods do not correspond to the nominal rates set on the devices. This paper describes a simple method to measure acceleration rate of the rotarod and to set the rate to a desired value for any brand of rod.
Genes, Brain and Behavior, 2012
It is known that four common inbred mouse strains show defects of the forebrain commissures. The ... more It is known that four common inbred mouse strains show defects of the forebrain commissures. The BALB/cJ strain has a low frequency of abnormally small corpus callosum, whereas the 129 strains have many animals with deficient corpus callosum. The I/LnJ and BTBR T+ tf/J strains never have a corpus callosum, whereas half of I/LnJ and almost all BTBR show severely reduced size of the hippocampal commissure. Certain F1 hybrid crosses among these strains are known to be less severely abnormal than the inbred parents, suggesting that the parent strains have different genetic causes of commissure defects. In this study, all hybrid crosses among the four strains were investigated. The BTBR × I/Ln hybrid expressed almost no defects of the hippocampal commissure, unlike its inbred parent strains. Numerous three-way crosses among the four strains yielded many mice with no corpus callosum and severely reduced hippocampal commissure, which shows that the phenotypic defect can result from several different combinations of genetic alleles. The F2 and F3 hybrid crosses of BTBR and I/LnJ had almost 100% absence of the corpus callosum but about 50% frequency of deficient hippocampal commissure. The four-way hybrid cross among all four abnormal strains involved highly fertile parents and yielded a very wide phenotypic range of defects from almost no hippocampal commissure to totally normal forebrain commissures. The F2 and F3 crosses as well as the four-way cross provide excellent material for studies of genetic linkage and behavioral consequences of commissure defects.
A central mesencephalic reticular formation projection to the supraoculomotor area in macaque monkeys
Brain structure & function, Jan 10, 2015
The central mesencephalic reticular formation is physiologically implicated in oculomotor functio... more The central mesencephalic reticular formation is physiologically implicated in oculomotor function and anatomically interwoven with many parts of the oculomotor system's premotor circuitry. This study in Macaca fascicularis monkeys investigates the pattern of central mesencephalic reticular formation projections to the area in and around the extraocular motor nuclei, with special emphasis on the supraoculomotor area. It also examines the location of the cells responsible for this projection. Injections of biotinylated dextran amine were stereotaxically placed within the central mesencephalic reticular formation to anterogradely label axons and terminals. These revealed bilateral terminal fields in the supraoculomotor area. In addition, dense terminations were found in both the preganglionic Edinger-Westphal nuclei. The dense terminations just dorsal to the oculomotor nucleus overlap with the location of the C-group medial rectus motoneurons projecting to multiply innervated musc...
Behav Brain Res., 2014
h i g h l i g h t s
Journal of Neuroscience Methods, 2009
The latency of mice and rats to fall from the accelerating rotarod can differ markedly between la... more The latency of mice and rats to fall from the accelerating rotarod can differ markedly between laboratories using the same brand of rod as well as between studies using different kinds of rods. These discrepancies can arise from different rod diameters, surface textures, test protocols, or laboratory environmental factors beyond the test itself, but it is also possible that the actual acceleration rates of the different rods do not correspond to the nominal rates set on the devices. This paper describes a simple method to measure acceleration rate of the rotarod and to set the rate to a desired value for any brand of rod.
Journal of Neuroscience Methods, 2010
- The precision of video and photocell tracking systems and the elimination of tracking error... more 2010) The precision of video and photocell tracking systems and the elimination of tracking errors with infrared backlighting. Journal of Neuroscience Methods, Made available courtesy of Elsevier: http://www.elsevier.com
Database for the design of tailored light delivery for optogenetics in non-human primates
This study analyzes the light emission pattern of tapered optical fibers (TFs) across different n... more This study analyzes the light emission pattern of tapered optical fibers (TFs) across different numerical apertures (NA), core diameters, laser input powers, cone angles, and light injection angles. The results will be used to tailor optical fibers for optimal light delivery to photosensitive neuronal actuators in the non-human primate (NHP) brain.
Superior colliculus projections to target populations in the supraoculomotor area of the macaque monkey
Visual Neuroscience, 2021
A projection by the superior colliculus to the supraoculomotor area (SOA) located dorsal to the o... more A projection by the superior colliculus to the supraoculomotor area (SOA) located dorsal to the oculomotor complex was first described in 1978. This projection’s targets have yet to be identified, although the initial study suggested that vertical gaze motoneuron dendrites might receive this input. Defining the tectal targets is complicated by the fact the SOA contains a number of different cell populations. In the present study, we used anterograde tracers to characterize collicular axonal arbors and retrograde tracers to label prospective SOA target populations in macaque monkeys. Close associations were not found with either superior or medial rectus motoneurons whose axons supply singly innervated muscle fibers. S-group motoneurons, which supply superior rectus multiply innervated muscle fibers, appeared to receive a very minor input, but C-group motoneurons, which supply medial rectus multiply innervated muscle fibers, received no input. A number of labeled boutons were observe...
Cerebellar projections to the macaque midbrain tegmentum: Possible near response connections
Visual Neuroscience, 2021
Since most gaze shifts are to targets that lie at a different distance from the viewer than the c... more Since most gaze shifts are to targets that lie at a different distance from the viewer than the current target, gaze changes commonly require a change in the angle between the eyes. As part of this response, lens curvature must also be adjusted with respect to target distance by the ciliary muscle. It has been suggested that projections by the cerebellar fastigial and posterior interposed nuclei to the supraoculomotor area (SOA), which lies immediately dorsal to the oculomotor nucleus and contains near response neurons, support this behavior. However, the SOA also contains motoneurons that supply multiply innervated muscle fibers (MIFs) and the dendrites of levator palpebrae superioris motoneurons. To better determine the targets of the fastigial nucleus in the SOA, we placed an anterograde tracer into this cerebellar nucleus in Macaca fascicularis monkeys and a retrograde tracer into their contralateral medial rectus, superior rectus, and levator palpebrae muscles. We only observed...
Adeno-Associated Virus Capsid-Promoter Interactions in the Brain Translate from Rat to the Nonhuman Primate
Human Gene Therapy, 2020
Recently, we established an AAV9 capsid-promoter interaction that directly determined cell-specif... more Recently, we established an AAV9 capsid-promoter interaction that directly determined cell-specific gene expression across two synthetic promoters, Cbh and CBA, in the rat striatum. The present studies not only expand this capsid-promoter interaction to include another promoter in the rat striatum, but also establishes AAV capsid-promoter interactions in the non-human primate brain. When AAV9 vectors were injected into the rat striatum, the minimal synthetic promoter JetI drove GFP gene expression predominantly in oligodendrocytes. However, similar to our previous findings, the insertion of 6 alanines into VP1/VP2 of the AAV9 capsid (AAV9AU) significantly shifted JetI-driven GFP gene expression to neurons. In addition, previous retrograde tracing studies in the non-human primate brain also revealed the existence of a capsid-promoter interaction. When rAAV2-Retro vectors were infused into the frontal eye field (FEF) of rhesus macaques, local gene expression was prominent using either the hybrid chicken beta actin (CAG) or human synapsin (hSyn) promoters. However, only the CAG promoter, not the hSyn promoter, led to gene expression in the ipsilateral claustrum and contralateral FEF. Conversely, infusion of rAAV2-retro-hSyn vectors, but not rAAV2-retro-CAG, into the macaque superior colliculus led to differential and selective retrograde gene expression in cerebellotectal afferent cells. Clearly, this differential promoter/capsid expression profile could not be attributed to promoter inactivation from retrograde transport of the rAAV2-Retro vector. In summary, we document the potential for AAV capsid/promoter interactions to impact cell specific gene expression across species, experimental manipulations and engineered capsids, independent of capsid permissivity.
Neuron, 2020
Highlights d Openly shared, non-human primate optogenetics data resource d Over 500 entries of un... more Highlights d Openly shared, non-human primate optogenetics data resource d Over 500 entries of unpublished data d Efficiency analysis of experimental parameters d Advice from leading NHP optogenetics labs
Frontiers in Neuroanatomy, 2019
Reliable viral vector-mediated transgene expression in primate motoneurons would improve our abil... more Reliable viral vector-mediated transgene expression in primate motoneurons would improve our ability to anatomically and physiologically interrogate motor systems. We therefore investigated the efficacy of replication defective, early region 1-deleted canine adenovirus type-2 (CAV-2) vectors for mediating transgene expression of fluorescent proteins into brainstem motoneurons following craniofacial intramuscular injections in four rhesus monkeys (Macaca mulatta). Vector injections were placed into surgically identified and isolated craniofacial muscles. After a 1-to 2-month survival time, animals were sacrificed and transgene expression was assessed with immunohistochemistry in the corresponding motoneuronal populations. We found that injections of CAV-2 into individual craniofacial muscles at doses in the range of ∼10 10 to 10 11 physical particles/muscle resulted in robust motoneuronal transduction and expression of immunohistochemically identified fluorescent proteins across multiple animals. By using different titers in separate muscles, with the resulting transduction patterns tracked via fluorophore expression and labeled motoneuron location, we established qualitative dose-response relationships in two animals. In one animal that received an atypically high titer (5.7 × 10 11 total CAV-2 physical particles) distributed across numerous injection sites, no transduction was detected, likely due to a retaliatory immune response. We conclude that CAV-2 vectors show promise for genetic modification of primate motoneurons following craniofacial intramuscular injections. Our findings warrant focused attention toward the use of CAV-2 vectors to deliver opsins, DREADDs, and other molecular probes to improve genetics-based methods for primate research. Further work is required to optimize CAV-2 transduction parameters. CAV-2 vectors encoding proteins could provide a new, reliable route for modifying activity in targeted neuronal populations of the primate central nervous system.
Journal of Comparative Psychology, 2019
Decisions often involve the consideration of multiple cues, each of which may inform selection on... more Decisions often involve the consideration of multiple cues, each of which may inform selection on the basis of learned probabilities. Our ability to use probabilistic inference for decisions is bounded by uncertainty and constraints such as time pressure. Previous work showed that when humans choose between visual objects in a multiple-cue, probabilistic task, they cope with time pressure by discounting the least informative cues, an example of satisficing or "good enough" decision-making. We tested two rhesus macaques (Macaca mulatta) on a similar task to assess their capacity for probabilistic inference and satisficing in comparison with humans. On each trial, a monkey viewed two compound stimuli consisting of four cue dimensions. Each dimension (e.g., color) had two possible states (e.g., red or blue) with different probabilistic weights. Selecting the stimulus with highest total weight yielded higher odds of receiving reward. Both monkeys learned the assigned weights at high accuracy. Under time pressure, both monkeys were less accurate as a result of decreased use of cue information. One monkey adopted the same satisficing strategy used by humans, ignoring the least informative cue dimension. Both monkeys, however, exhibited a strategy not reported for humans, a "group-the-best" strategy in which the top two cues were used similarly despite their different assigned weights. The results validate macaques as an animal model of probabilistic decision-making, establishing their capacity to discriminate between objects using at least four visual dimensions simultaneously. The time pressure data suggest caution, however, in using macaques as models of human satisficing.
The Anatomical Record, 2019
The organization of extraocular muscles (EOMs) and their motor nuclei was investigated in the mou... more The organization of extraocular muscles (EOMs) and their motor nuclei was investigated in the mouse due to the increased importance of this model for oculomotor research. Mice showed a standard EOM organization pattern, although their eyes are set at the side of the head. They do have more prominent oblique muscles, whose insertion points differ from those of frontal-eyed species. Retrograde tracers revealed that the motoneuron layout aligns with the general vertebrate plan with respect to nuclei and laterality. The mouse departed in some significant respects from previously studied species. First, more overlap between the distributions of muscle-specific motoneuronal pools was present in the oculomotor nucleus (III). Furthermore, motoneuron dendrites for each pool filled the entire III and extended beyond the edge of the abducens nucleus (VI). This suggests mouse extraocular motoneuron afferents must target specific pools based on features other than dendritic distribution and nuclear borders. Second, abducens internuclear neurons are located outside the VI. We concluded this because no unlabeled abducens internuclear neurons were observed following lateral rectus muscle injections and because retrograde tracer injections into the III labeled cells immediately ventral and ventrolateral to the VI, not within it. This may provide an anatomical substrate for differential input to motoneurons and internuclear neurons that This article includes AR WOW Video. Video can be viewed at https://players.brightcove.net/656326989001/default_default/ index.html?videoId=6032629801001 Abbreviations: 4V = fourth ventricle; aVI = accessory abducens nucleus; BC = brachium conjunctivum; CA = cerebral aqueduct; CG = central gray; DR = dorsal raphe; EWcp = centrally projecting Edinger-Westphal nucleus; III = oculomotor nucleus; IO = inferior oblique muscle; IV = trochlear nucleus; LP = levator palpebrae superioris muscle; LR = lateral rectus muscle; MLF = medial longitudinal fasciculus; MR = medial rectus muscle; MRF = mesencephalic reticular formation; ON = optic nerve;
Molecular and Cellular Endocrinology, 2017
Three forms of familial primary aldosteronism have been recognized. Familial Hyperaldosteronism t... more Three forms of familial primary aldosteronism have been recognized. Familial Hyperaldosteronism type 1 (FH1) or dexamethasone suppressible hyperaldosteronism, FH2, the most common form of as yet unknown cause(s), and FH3. FH3 is due to activating mutations of the potassium channel gene KCNJ5 that increase constitutive and angiotensin II-induced aldosterone synthesis. In this study we examined the cellular distribution of CYP11B2, CYP11B1, CYP17A1 and KCNJ5 in adrenals from two FH3 siblings using immunohistochemistry and immunofluorescence and obtained unexpected results. The adrenals were markedly enlarged with loss of zonation. CYP11B2 was expressed sporadically throughout the adrenal cortex. CYP11B2 was most often expressed by itself, relatively frequently with CYP17A1, and less frequently with CYP11B1. KCNJ5 was co-expressed with CYP11B2 and in some cells with CYP11B1. This aberrant co-expression of enzymes likely explains the abnormally high secretion rate of the hybrid steroid, 18-oxocortisol.
The Journal of comparative neurology, Jan 2, 2016
In this study, two points related to the pattern of innervation of the extraocular muscles were e... more In this study, two points related to the pattern of innervation of the extraocular muscles were explored. 1. Species differences exist in the location of the motoneurons supplying multiply innervated fibers (MIFs) and singly innervated fibers (SIFs) in eye muscles. MIF motoneurons are located outside the extraocular nuclei in primates, but are intermixed with SIF motoneurons within rat extraocular nuclei. To test whether this difference is related to visual capacity and frontal placement of eyes, we injected retrograde tracers into the medial rectus muscle of the cat, a highly visual, non-primate with frontally placed eyes. Distal injections labeled smaller MIF motoneurons located ventrolaterally and rostrally within the oculomotor nucleus (III). More central injections also labeled a separate population of larger cells, located dorsally in III. Thus, the cat shares the feature of having MIF motoneurons located within the bounds of III with the nocturnal rat. On the other hand, like...
Journal of AAPOS : the official publication of the American Association for Pediatric Ophthalmology and Strabismus / American Association for Pediatric Ophthalmology and Strabismus, 2016
Extraocular proprioception has been shown to participate in spatial perception and binocular alig... more Extraocular proprioception has been shown to participate in spatial perception and binocular alignment. Yet the physiological approaches used to study this sensory signal are limited because proprioceptive signaling takes place at the same time as visuomotor signaling. It is critical to dissociate this sensory signal from other visuomotor events that accompany eye movements. We present a novel noninvasive and quantifiable method for probing extraocular proprioception independent of other visuomotor processing by attaching a rare-earth magnet to a real-time model eye and placing an electromagnet <20 mm from the eye. An electromagnet can increase or decrease angular displacements and velocities of the model eye. Electromagnetic activation rapidly affected (<2 ms) the rotation kinematics of the eye, which were correlated linearly with both the current supply and the distance of the electromagnet relative to the eye. This method circumvented the constraints of conventional physiol...
Physiology & behavior, Jan 11, 2015
Exposure to unpredictable chronic mild stress (CUS) is a commonly used protocol in rats that is r... more Exposure to unpredictable chronic mild stress (CUS) is a commonly used protocol in rats that is reported to evoke antidepressant-reversible behaviors such as loss of preference for sweetened water solution which is taken as an analog of the anhedonia seen in major depression. However, the induction of anhedonic-like behavior by chronic mild stress, gauged by an animal's preference for sucrose solution, is not fully reproducible and consistent across laboratories. In this study, we compared a widely used behavioral marker of anhedonia - the sucrose preference test, with another phenotypic marker of emotional valence, social interaction-associated ultrasonic vocalizations as well as a marker of an anxiety-like phenotype, novelty-suppressed feeding, and cognitive performance in the eight arm radial maze task in adult male Sprague-Dawley rats. Chronic four-week exposure to unpredictable mild stressors resulted in 1) attenuation of social interaction-associated ultrasonic vocalizatio...
Using rAAV2-retro in rhesus macaques: Promise and caveats for circuit manipulation
Journal of Neuroscience Methods, 2020
BACKGROUND Recent genetic technologies such as opto- and chemogenetics allow for the manipulation... more BACKGROUND Recent genetic technologies such as opto- and chemogenetics allow for the manipulation of brain circuits with unprecedented precision. Most studies employing these techniques have been undertaken in rodents, but a more human-homologous model for studying the brain is the nonhuman primate (NHP). Optimizing viral delivery of transgenes encoding actuator proteins could revolutionize the way we study neuronal circuits in NHPs. NEW METHOD: rAAV2-retro, a popular new capsid variant, produces robust retrograde labeling in rodents. Whether rAAV2-retro's highly efficient retrograde transport would translate to NHPs was unknown. Here, we characterized the anatomical distribution of labeling following injections of rAAV2-retro encoding opsins or DREADDs in the cortico-basal ganglia and oculomotor circuits of rhesus macaques. RESULTS rAAV2-retro injections in striatum, frontal eye field, and superior colliculus produced local labeling at injection sites and robust retrograde labeling in many afferent regions. In every case, however, a few brain regions with well-established projections to the injected structure lacked retrogradely labeled cells. We also observed robust terminal field labeling in downstream structures. COMPARISON WITH EXISTING METHOD(S) Patterns of labeling were similar to those obtained with traditional tract-tracers, except for some afferent labeling that was noticeably absent. CONCLUSIONS rAAV2-retro promises to be useful for circuit manipulation via retrograde transduction in NHPs, but caveats were revealed by our findings. Some afferently connected regions lacked retrogradely labeled cells, showed robust axon terminal labeling, or both. This highlights the importance of anatomically characterizing rAAV2-retro's expression in target circuits in NHPs before moving to manipulation studies.
Behav Brain Res., 2014
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Journal of Neuroscience Methods, 2009
The latency of mice and rats to fall from the accelerating rotarod can differ markedly between la... more The latency of mice and rats to fall from the accelerating rotarod can differ markedly between laboratories using the same brand of rod as well as between studies using different kinds of rods. These discrepancies can arise from different rod diameters, surface textures, test protocols, or laboratory environmental factors beyond the test itself, but it is also possible that the actual acceleration rates of the different rods do not correspond to the nominal rates set on the devices. This paper describes a simple method to measure acceleration rate of the rotarod and to set the rate to a desired value for any brand of rod.
Genes, Brain and Behavior, 2012
It is known that four common inbred mouse strains show defects of the forebrain commissures. The ... more It is known that four common inbred mouse strains show defects of the forebrain commissures. The BALB/cJ strain has a low frequency of abnormally small corpus callosum, whereas the 129 strains have many animals with deficient corpus callosum. The I/LnJ and BTBR T+ tf/J strains never have a corpus callosum, whereas half of I/LnJ and almost all BTBR show severely reduced size of the hippocampal commissure. Certain F1 hybrid crosses among these strains are known to be less severely abnormal than the inbred parents, suggesting that the parent strains have different genetic causes of commissure defects. In this study, all hybrid crosses among the four strains were investigated. The BTBR × I/Ln hybrid expressed almost no defects of the hippocampal commissure, unlike its inbred parent strains. Numerous three-way crosses among the four strains yielded many mice with no corpus callosum and severely reduced hippocampal commissure, which shows that the phenotypic defect can result from several different combinations of genetic alleles. The F2 and F3 hybrid crosses of BTBR and I/LnJ had almost 100% absence of the corpus callosum but about 50% frequency of deficient hippocampal commissure. The four-way hybrid cross among all four abnormal strains involved highly fertile parents and yielded a very wide phenotypic range of defects from almost no hippocampal commissure to totally normal forebrain commissures. The F2 and F3 crosses as well as the four-way cross provide excellent material for studies of genetic linkage and behavioral consequences of commissure defects.
A central mesencephalic reticular formation projection to the supraoculomotor area in macaque monkeys
Brain structure & function, Jan 10, 2015
The central mesencephalic reticular formation is physiologically implicated in oculomotor functio... more The central mesencephalic reticular formation is physiologically implicated in oculomotor function and anatomically interwoven with many parts of the oculomotor system's premotor circuitry. This study in Macaca fascicularis monkeys investigates the pattern of central mesencephalic reticular formation projections to the area in and around the extraocular motor nuclei, with special emphasis on the supraoculomotor area. It also examines the location of the cells responsible for this projection. Injections of biotinylated dextran amine were stereotaxically placed within the central mesencephalic reticular formation to anterogradely label axons and terminals. These revealed bilateral terminal fields in the supraoculomotor area. In addition, dense terminations were found in both the preganglionic Edinger-Westphal nuclei. The dense terminations just dorsal to the oculomotor nucleus overlap with the location of the C-group medial rectus motoneurons projecting to multiply innervated musc...
Behav Brain Res., 2014
h i g h l i g h t s
Journal of Neuroscience Methods, 2009
The latency of mice and rats to fall from the accelerating rotarod can differ markedly between la... more The latency of mice and rats to fall from the accelerating rotarod can differ markedly between laboratories using the same brand of rod as well as between studies using different kinds of rods. These discrepancies can arise from different rod diameters, surface textures, test protocols, or laboratory environmental factors beyond the test itself, but it is also possible that the actual acceleration rates of the different rods do not correspond to the nominal rates set on the devices. This paper describes a simple method to measure acceleration rate of the rotarod and to set the rate to a desired value for any brand of rod.
Journal of Neuroscience Methods, 2010
- The precision of video and photocell tracking systems and the elimination of tracking error... more 2010) The precision of video and photocell tracking systems and the elimination of tracking errors with infrared backlighting. Journal of Neuroscience Methods, Made available courtesy of Elsevier: http://www.elsevier.com