Interdigitated Paralemniscal and Lemniscal Pathways in the Mouse Barrel Cortex

Primary sensory cortical areas receive information through multiple thalamic channels. In the rodent whisker system, lemniscal and paralemniscal thalamocortical projections, from the ventral posteromedial nucleus (VPM) and posterior medial nucleus (POm) respectively, carry distinct types of sensory information to cortex. Little is known about how these separate streams of activity are parsed and integrated within the neocortical microcircuit. We used quantitative laser scanning photostimulation to probe the organization of functional thalamocortical and ascending intracortical projections in the mouse barrel cortex. To map the thalamocortical projections, we recorded from neocortical excitatory neurons while stimulating VPM or POm. Neurons in layers (L)4, L5, and L6A received dense input from thalamus (L4, L5B, and L6A from VPM; and L5A from POm), whereas L2/3 neurons rarely received thalamic input. We further mapped the lemniscal and paralemniscal circuits from L4 and L5A to L2/3. Lemniscal L4 neurons targeted L3 within a column. Paralemniscal L5A neurons targeted a superficial band (thickness, 60 μm) of neurons immediately below L1, defining a functionally distinct L2 in the mouse barrel cortex. L2 neurons received input from lemniscal L3 cells and paralemniscal L5A cells spread over multiple columns. Our data indicate that lemniscal and paralemniscal information is segregated into interdigitated cortical layers

Spatiotemporal Properties of the Action Potential Propagation in the Mouse Visual Cortical Slice Analyzed by Calcium Imaging

The calcium ion (Ca2+) is an important messenger for signal transduction, and the intracellular Ca2+ concentration ([Ca2+]i) changes in response to an excitation of the cell. To reveal the spatiotemporal properties of the propagation of an excitatory signal with action potentials in the primary visual cortical circuit, we conducted a Ca2+ imaging study on slices of the mouse visual cortex. Electrical stimulation of layer 4 evoked [Ca2+]i transients around the stimulus electrode. Subsequently, the high [Ca2+]i region mainly propagated perpendicular to the cortical layer (vertical propagation), with horizontal propagation being restricted. When the excitatory synaptic transmission was blocked, only weak and concentric [Ca2+]i transients were observed. When the action potential was blocked, the [Ca2+]i transients disappeared almost completely. These results suggested that the action potential contributed to the induction of the [Ca2+]i transients, and that excitatory synaptic connections were involved in the propagation of the high [Ca2+]i region in the primary visual cortical circuit. To elucidate the involvement of inhibitory synaptic connections in signal propagation in the primary visual cortex, the GABAA receptor inhibitor bicuculline was applied. In this case, the evoked signal propagated from layer 4 to the entire field of view, and the prolonged [Ca2+]i transients were observed compared with the control condition. Our results suggest that excitatory neurons are widely connected to each other over the entire primary visual cortex with recurrent synapses, and inhibitory neurons play a fundamental role in the organization of functional sub-networks by restricting the propagation of excitation signals

Absence of Gal epitope prolongs survival of swine lungs in an ex vivo model of hyperacute rejection

Background: Galactosyl transferase gene knock-out (GalTKO) swine offer a unique tool to evaluate the role of the Gal antigen in xenogenic lung hyperacute rejection. Methods: We perfused GalTKO miniature swine lungs with human blood. Results were compared with those from previous studies using wild-type and human decay-accelerating factor-transgenic (hDAF+/+) pig lungs. Results: GalTKO lungs survived 132 ± 52 min compared to 10 ± 9 min for wild-type lungs (P = 0.001) and 45 ± 60 min for hDAF+/+ lungs (P = 0.18). GalTKO lungs displayed stable physiologic flow and pulmonary vascular resistance (PVR) until shortly before graft demise, similar to autologous perfusion, and unlike wild-type or hDAF+/+ lungs. Early (15 and 60 min) complement (C3a) and platelet activation and intrapulmonary platelet deposition were significantly diminished in GalTKO lungs relative to wild-type or hDAF+/+ lungs. However, GalTKO lungs adsorbed cytotoxic anti-non-Gal antibody and elaborated high levels of thrombin; their demise was associated with increased PVR, capillary congestion, intravascular thrombi and strong CD41 deposition not seen at earlier time points. Conclusions: In summary, GalTKO lungs are substantially protected from injury but, in addition to anti-non-Gal antibody and complement, platelet adhesion and non-physiologic intravascular coagulation contribute to Gal-independent lung injury mechanisms. © 2011 John Wiley & Sons A/S

Protective effects of 5-HT(1A) receptor agonists against emotional changes produced by stress stimuli are related to their neuroendocrine effects

1. The effects of the 11β-hydroxylase inhibitor metyrapone on the protective effects of serotonin (5-hydroxytryptamine; 5-HT)(1A) receptor agonists against emotional changes produced by acute restraint stress were examined in mice. 2. Changes in the emotional state of mice were evaluated in terms of changes in exploratory activity, i.e. total locomotor activity, number and duration of rearing and head-dipping behaviours, and latency to the first head-dipping, using an automatic hole-board apparatus. 3. Treatment with the 5-HT(1A) receptor agonists flesinoxan (1 mg kg(−1), i.p.) and R(+)-2-di-n-propylamino-8-hydroxy-1,2,3,4-tetrahydronaphthalene hydrobromide (8-OH-DPAT; 1 mg kg(−1), i.p.) 24 h prior to exposure to stress significantly suppressed the decrease in various exploratory behaviours that was observed immediately after the exposure to acute restraint stress (60 min). The effects of flesinoxan (1 mg kg(−1), i.p.) and 8-OH-DPAT (1 mg kg(−1), i.p.) were antagonized by co-injection with N-{2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl}-N-(2-pyridinyl) cyclohexanecarboxamide trihydrochloride (WAY100635; 1 mg kg(−1), i.p.), a selective 5-HT(1A) receptor antagonist. 4. Flesinoxan (1 mg kg(−1), i.p.) and 8-OH-DPAT (1 mg kg(−1) i.p.) significantly increased the plasma corticosterone level, and these effects of 5-HT(1A) receptor agonists were dose-dependently blocked by pretreatment with metyrapone (12.5 and 25 mg kg(−1), s.c.). 5. Metyrapone (25 mg kg(−1), s.c.) alone did not modify the stress-induced changes in exploratory behaviours. Pretreatment with metyrapone (12.5 and 25 mg kg(−1), s.c.) partly antagonized the protective effects of flesinoxan (1 mg kg(−1), i.p.) and 8-OH-DPAT (1 mg kg(−1), i.p.) with regard to only the number and duration of head-dipping behaviours. 6. These results suggest that activation of the adrenocortical system via 5-HT(1A) receptors may facilitate some adaptive mechanism(s) involved in the recognition of and/or ability to cope with stressful situations