Original recordings were then decimated (averaged 10 points, 1 ms). Single peak spontaneous IPSCs with amplitudes greater than 2.1 times the SD of baseline noise were detected using a semiautomated sliding template detection procedure with AxoGraph X. The template was generated by averaging

multiple sIPSCs. Each detected event was visually inspected. Events were discarded if the average baseline noise (>300 ms) was greater than the peak ±1.5 s from the peak. Peak amplitudes were determined by averaging the current ±20 ms this website from the greatest upward deflection. The amplitude distribution of the baseline noise was measured by averaging the baseline current ±20 ms from the greatest upward deflection, 220 ms after a point set to 0 pA, once every 50 s (n = 26 cells). To compare the kinetics of eIPSCs to sIPSCs, spontaneous events with a single peak were selected. Duration of eIPSCs and sIPSCs was determined by measuring the width at 20% of the peak amplitude (see Figure 1C). All drugs were applied through

bath perfusion, except dopamine, which was applied by iontophoresis. Iontophoretic pipettes (70–110 MΩ) were filled with 1 M dopamine and the tip placed within 10 μm of the soma. A negative backing current (6–11 nA) prevented leakage. Dopamine was ejected with the application of positive current (2–6 s) with an Axoclamp 2A amplifier to elicit a maximal dopamine-induced outward current. A transgenic mouse expressing the human D2 dopamine receptor short isoform (hD2S) with a flag CB-839 mouse epitope on the amino terminus was generated by nuclear microinjection using standard techniques. The transgene consisted of an 8.5 kb genomic fragment from the rat tyrosine hydroxylase gene (TH) containing 5′ regulatory sequences, the basal promoter, and 26 base pairs from the 5′ untranslated region in exon 1 followed by a 0.7 kb cassette containing intron 2 and splice donor/acceptor sites from the rabbit beta-globin gene (Arttamangkul et al., 2008). The hD2S construct consisted of a consensus Kozak sequence, a signal peptide from the hemagglutinin

influenza followed by the sequence for the FLAG epitope (Vickery and von Zastrow, 1999), a full-length cDNA for the hD2S containing 1.4 kb of coding Cediranib (AZD2171) sequence and 1.0 kb of 3′ untranslated, and the bovine growth hormone polyA sequence from pcDNA 3.0 (Invitrogen). After cleavage by the signal peptidase of the signal sequence during translation, an hD2S protein with an amino terminus Flag epitope is expressed in TH-expressing neurons including the dopamine neurons of the midbrain, as shown by immunostaining with the M1 anti-Flag antibody (Sigma-Aldrich) using confocal microscopy on sections and two-photon microscopy on midbrain slice preparations (data not shown). CGP 35348 and 5-CT were obtained from Tocris Bioscience. Baclofen and sulpiride were obtained from Research Biochemical. MK-801 was obtained from Abcam.

, 1992). Within fungi, the potential for antibiotic production is also an undesired property. The occurrence of virulence traits should not be present in microorganisms used in food fermentation. A specific risk assessment should be conducted on strains presenting these undesirable properties, even if they belong to a species with a long history of use (Semedo et al., 2003a and Semedo et al., 2003b). The emergence

and spread of antibiotic resistance is a major global health concern. The on-going Codex ad hoc intergovernmental task force on antimicrobial resistance is focused on the non-human use of antimicrobials. Microorganisms intentionally added to food and feed for technological purposes have not been shown to aggravate the problem of spreading antibiotic resistant pathogens Vemurafenib (Anon, 2001). Intrinsic resistance or resistance that is caused by mutation in an indigenous gene not associated with mobile elements would represent a very low risk of dissemination (Saarela et al., 2007). Acquired antibiotic resistance genes, especially when associated with mobile genetic elements (plasmids, transposons), can be transferred to pathogens or other commensals along the food chain, from within the product until consumption (FEEDAP, 2005, FEEDAP, 2008 and Nawaz et al., 2011). The role of MFC in the spread of antibiotic

resistance has been assessed in fermented foods (Nawaz et al., 2011) as well as

more specifically for probiotic food products (Saarela Dolutegravir concentration et al., 2007, Mater et al., 2008 and Vankerckhoven et al., 2008). Results of such studies confirm the role of a reservoir of antibiotic resistance genes from the food microbiota, without identifying any major health concerns to date. It is considered Dabigatran that strains carrying acquired antibiotic resistance genes might act as a reservoir of transmissible antimicrobial resistance determinants (FEEDAP, 2005 and FEEDAP, 2008). Gene transfer of antibiotic resistance between microorganisms in the food and feed chain is thus considered to be a topic of surveillance for the safety demonstration of microorganisms (FAO and WHO, 2001, FAO and WHO, 2002, Borriello et al., 2003 and Gueimonde et al., 2005). The “2002 IDF Inventory” listed 82 bacterial species and 31 species of yeast and molds whereas the present “Inventory of MFC” contains 195 bacterial species and 69 species of yeasts and molds. The overview of the distribution of species over the relevant taxonomic units is given in Table 1 for bacteria and Table 2 and Table 3 for fungi. We publish the complete current “Inventory of Microbial Food Cultures” as accompanying material to the present paper. The genus Brachybacterium enters the list with two species, B. alimentarium and B. tyrofermentans.

We first examined whether Sema3A serves as a polarizing factor for axon/dendrite differentiation in cultured hippocampal neurons (Dotti and Banker, 1987 and Dotti et al., 1988). For comparison, we also tested the effect of netrin-1, BDNF, and NGF, secreted factors known to be involved in neuronal polarization in various systems. Dissociated hippocampal neurons from rat embryos were plated on substrates

coated with stripes (50 μm wide with 50 μm gap) of the recombinant form of Sema3A, BDNF, NGF, or netrin-1 (see Experimental Procedures). To examine neuronal polarization, we imaged neurons at 12 and 60 hr after cell plating, before and after axon/dendrite differentiation, respectively. At 12 hr, the cells exhibited several short neurites of similar lengths without selleck kinase inhibitor apparent

polarity (Figure 1A), whereas most cells developed a single axon and multiple dendrites at 60 hr, as shown by immunostaining with axonal marker Smi-312 and somatodendritic marker MAP2 (Figure 1A). Strikingly, we found that axons were mostly formed off the Sema3A-coated stripe, whereas more dendrites were found to differentiate on than off the Sema3A stripe ( Figure 1A). Furthermore, axonal growth cones often turned at the stripe boundary to stay away from the Sema3A stripe, whereas dendrites showed opposite tendency ( Figure 1A), suggesting Enzalutamide attractive and repulsive actions of Sema3A on dendritic and axonal growth cones, respectively. The effect of Sema3A on axon/dendrite formation was quantified by determining the distribution of axon/dendrite initiation sites on the soma for all polarized cells with their somata located on the stripe boundary at 48–60 hr, when neurons had completed the polarization process (Figures 1Ba and 1Bb). Because the neurite initiation site on the soma does not move significantly during axon/dendrite differentiation (Figure 1A), this retrospective analysis allowed us to determine whether coated stripes influenced axon/dendrite Phosphatidylinositol diacylglycerol-lyase differentiation after neurites had been initiated from the soma. We found

that axon differentiation largely occurred for neurites initiated off the Sema3A stripe, whereas slightly more dendrites developed on the Sema3A stripe ( Figure 1Bb). We also found that the preference of axon/dendrite formation on BDNF-coated stripes was opposite to that for Sema3A stripes ( Figure 1Bb), consistent with a previous report ( Shelly et al., 2007). In contrast, we found no preference of axon/dendrite differentiation for stripes coated with BSA or NGF, and a slight preference of dendrite differentiation away from the netrin-1 stripes ( Figure 1Bb). In Figure 1Ca, these results on axon/dendrite formation are quantified by using the preference index (PI = [(% on stripe) − (% off stripe)] / 100%). Overall, the most striking effect of Sema3A on neuronal polarization is its suppression of axon differentiation, resulting in strong preference of axon formation away from Sema3A stripes ( Figures 1Bb and 1Ca).

, 2010). In mice, transplantation of embryonic cells can enhance GDC-0973 supplier the critical-period plasticity of the visual cortex (Southwell et al., 2010). A decade of preclinical research into the use of adult and fetal/progenitor cells in animal models of ischemic stroke (Bliss et al., 2007, Leong et al., 2013 and Sanberg et al., 2012) showed that transplanted cells may act through the secretion of soluble factors that promote neurogenesis, angiogenesis, and immunomodulation (Leong et al., 2013). Although much has to be understood regarding efficacy

and mechanisms of action, there are now multiple ongoing early-phase clinical trials using cell-based therapies in stroke patients (Misra et al., 2012). Invasive and noninvasive electrical stimulation may modulate neural circuits in a wide range of disease states and allow recovery of normal circuit functions (Demirtas-Tatlidede et al., 2013, Hallett, 2000, Holtzheimer and Mayberg, 2011, Hsu et al., 2012, Kuo et al., 2013, Nitsche and Paulus, 2000 and Perlmutter and Mink, 2006). As outlined above, DBS has rapidly emerged as an important therapeutic tool in movement disorders as well as other neurological and psychiatric diseases, although the precise underlying physiological LGK-974 purchase mechanisms need to be clarified. Noninvasive electrical stimulation of large cortical areas could be

achieved by transcranial magnetic stimulation (TMS) that depends on the induction of electrical currents via externally applied magnetic

fields, or by transcranial direct current stimulation (tDCS) based on the penetration of externally applied electrical currents through the skull. Multiple studies have shown that both TMS and tDCS can impact motor and cognitive functions in healthy subjects and patients with neurological or psychiatric disorders (Demirtas-Tatlidede et al., 2013, Hsu et al., 2012, Hummel et al., 2005 and Kuo et al., 2013). TMS is currently approved for medication-refractory depression (Demirtas-Tatlidede et al., 2013). In stroke, both tDCS and repetitive TMS over the injured Sodium butyrate hemisphere when paired with training can improve motor performance and facilitate motor recovery (Grefkes and Fink, 2012 and Hsu et al., 2012). Stimulation-induced activity-dependent synaptic plasticity appears to be a potential mechanism of action. For example, an in vitro study found that both NMDA-R activation and BDNF are required for induction of synaptic potentiation via direct current stimulation that mimicked tDCS (Fritsch et al., 2010). Early work by Fetz and colleagues laid the foundation for real-time processing of neural signals and the induction of neural plasticity through feedback (Fetz, 2007). For example, precisely timed microstimulation of an M1 cortical neuron using the spiking signal of an adjacent recorded “presynaptic” neuron over a period of 2 days resulted in a reorganization of the motor output in a manner resembling STDP-like synaptic potentiation (Jackson et al., 2006).

The frequency of the

FAMACHA© correct interpretations were verified by comparing the results obtained with the PCV of each animal. The proportion of success was based on the following PCV references: FAMACHA© score 1 (F1): values ≥28%, F2: 23–27%, F3: 18–22%, F4: 13–17%, and F5: ≤12%. To evaluate the correlation between the breeds of goats and the chart reading success, the 95% confidence interval was determined by Pearson. Means and standard errors (S.E.) for the counting of eggs per gram (EPG) of faeces were calculated for dairy goats in each FAMACHA© score category. The percentage of success on the interpretation of FAMACHA© ranged from 32.6% (May 2009) to 87.5% (April 2010). It was observed a low percentage of correctness for the use of buy Ion Channel Ligand Library FAMACHA© during the first three months with 32.6, 47.3 and 68%, respectively. INK1197 The percentage reached a considerable level above 70% after the fourth month of evaluation. The quarter success mean were 49.3% (first quarter), 76% (second quarter), 83.6% (third quarter) and 76.1%

(fourth quarter). The most prevalent helminth in all larval culture was Haemonchus sp. (80.1%) followed by Trichostrongylus sp. (13.2%) and Oesophagostomum sp. (6.7%). Means and standard errors of EPG were calculated for dairy goats in each FAMACHA© score category (Table 1). There was an increase in the EPG values following the increase in the FAMACHA© score. More than half of the evaluated animals (52%) were classified as FAMACHA© score 2, with a mean of 515 EPG. The results confirm the objective of the

method in a semi-arid area of Brazil for not to deworm animals with FAMACHA© scores 1 and 2. Correlations between the variables are listed in Table 2. Although there was a good correlation between EPG and FAM there was no difference between seasons/months. PCV and FAMACHA were moderately negative correlated throughout the year as EPG and PCV. The occurrence of such finding may be related with the resilience characteristic of the breed in the region and the adjustment of the FAMACHA into goats. The factors EPG and FAMACHA and H. contortus had a much lower correlation (0.375) during the rainy season as compared to 0.707 PRKACG during the dry months. The correlation of treatment, the incidence of H. contortus, and rainfall was higher during the rainy season. The success of the evaluation rate after the first quarter (mean of 78.5%) are similar to those found by Chagas et al. (2007), who tested the FAMACHA© method in sheep herds in southeastern Brazil. We got a low success rate at the beginning of the use of the FAMACHA© method on the first quarter (mean of 55%) while in the second quarter, the success mean rose to 80%. Molento et al. (2004) attributed the low success rates at the start of the implementation due to the lack of experience of the observers in the conjunctiva colour interpretation as well as the use of the method in goats.

This requires new partnership models for research in which traditional silos are broken down, translational teams are created, and new mechanisms for effective hand-off from nonprofit to for-profit are generated. Today many researchers in the stem cell field have advanced their research far enough to attempt clinical translation but lack the knowledge and wherewithal to accomplish this arduous, expensive, and long-term task (Figure 1).

The significant hurdles needed to be surmounted are illustrated in the analysis of the drug development process (Figure 2). Despite these difficulties, steady progress toward this goal is being made, spearheaded by industry, academic institutions, and nonprofit foundations in conjunction with a recent focus by the National Institutes of Health (NIH) and the Food and Drug Administration (FDA) in the U.S. on both translational research and regenerative medicine. see more Here we describe the current status of, and

pathways for, stem cell-based CNS therapies, analyze the landscape of current regulatory approved clinical trials, discuss the recent industry trends and regulatory developments that can catalyze further translational progress, and describe key issues and currently available resources selleck inhibitor to facilitate more efficient translation of promising research. NSCs are the fundamental ancestor cells for the CNS (brain, spinal cord, and retina), defined by their ability to self-renew and produce all three major CNS cell types: neurons, astrocytes, and oligodendrocytes. NSCs can be expanded substantially, proliferating to produce cell lines that can differentiate into functional neural cells after in vivo transplantation, 4��8C demonstrating tremendous promise for cell replacement and regenerative

therapies. NSCs are abundant in different regions of the fetal CNS and are retained throughout life in restricted parts of the forebrain, notably the striatal subventricular zone and dentate gyrus of the hippocampus. Human NSCs have been isolated from donated fetal CNS tissue and can be defined by expression of surface markers such as CD133 (Uchida et al., 2000), enabling prospective enrichment, in vitro expansion using growth factors such as FGF2 and EGF, and in-depth characterization. NSC primary cell lines generated from human fetal CNS tissue, typically around 8–18 weeks of gestation, are now the subject of a number of clinical studies. Progenitor cells that arise from human NSCs, such as glial-restricted progenitor cells (GRPs), which produce oligodendrocytes and new myelin, are also being advanced toward the clinic (Goldman, 2011 and Sandrock et al., 2010). Other sources of neural cells showing promise in preclinical studies include cells from nasal mucosa such as olfactory ensheathing cells (Lindsay et al., 2010 and Raisman and Li, 2007) and skin-derived multipotent precursors (SKPs) (Fernandes et al., 2008).

To distinguish whether directional selectivity

in the presence of inhibitory blockers arose pre- or postsynaptically, the properties of ganglion cell light-evoked synaptic inputs were analyzed using whole-cell voltage-clamp techniques. In these experiments, after measuring spikes in cell-attached mode in the presence of blockers (Figure 6A), the same cell was patched with an electrode containing intracellular solution. After learn more break-in, the DSGC was dialyzed with QX314 and Cs+ and repeatedly injected with brief depolarizing pulses (−60–0 mV) until Na+ currents and a large fraction of voltage-gated K+ currents were blocked. Under these conditions, moving spots elicited large inward currents in both the null and preferred directions (VHOLD = −60 mV; Figure 6B). When the cell was held ∼0 mV, the inhibitory inputs that are usually associated with stimulating these cells (Figures 3A and S3) were not apparent, confirming that they were effectively blocked with the cocktail of antagonists (also see Figure S4). At +40 mV, light evoked outward currents. Importantly, the

temporal characteristics of currents measured at −60 and +40 mV were similar (Figure S5), indicating that they were not contaminated by voltage-dependent conductances, and thus provided a reliable readout of bipolar cell output. Reversal of the excitatory currents also indicated that gap junctions did not significantly contribute to the synaptic responses (Ackert et al., Akt inhibitor 2009). Under conditions in which inhibitory receptors and active postsynaptic conductances were blocked, preferred and null-direction stimuli evoked excitatory currents that were similar in size. The amplitude of the peak currents was not significantly different

whether measured at −60 mV (preferred: −228 ± 30 pA and −136 ± new 24 pA, for ON and OFF, respectively; null: −206 ± 30 pA and −131 ± 18 pA for ON and OFF, respectively; p > 0.6; n = 6) or +40 mV (preferred: 353 ± 64 pA and 214 ± 44 pA, for ON and OFF, respectively; null: 373 ± 67 pA and 216 ± 40 pA for ON and OFF, respectively; p > 0.6; n = 6; Figure 6C). Similarly, the total charge of the response was similar in magnitude in the null and preferred directions, indicating that moving spots stimulated an equal number of inputs in both directions (−60 mV ON, −113 ± 18 nC for preferred compared to −126 ± 21 nC for null; −60 mV OFF, −54 ± 13 nC for preferred compared to −66 ± 16 nC for null; +40 mV ON, 227 ± 52 nC for preferred compared to 265 ± 54 nC for null; +40 mV OFF, 124 ± 26 nC for preferred compared to 141 ± 29 nC for null; p > 0.5; n = 6; Figure 6D). The symmetry in input strength contrasts with the asymmetry in the spiking responses and suggests that nonlinearities within the ganglion cell must contribute to direction discrimination.

Because nicotine solutions have a bitter

taste, nicotine was diluted in saccharin solution, and control experiments were performed with this website a bitter solution (containing quinine). There were no differences in consumption of regular, sweetened, or bitter water between the two groups (Figure 6A). Next, we performed a free choice consumption experiment where mice were allowed to choose between regular water and water supplemented with different concentrations of nicotine (1–100 μg/ml) without saccharin. Analysis of the nicotine volume consumed relative to the total fluid intake (Figure 6C) indicated that Tabac mice significantly avoided drinking nicotine solutions containing more than 5 μg/ml nicotine (p < 0.05, two-way ANOVA),

while WT mice showed no preference between water and nicotine solutions below 50 μg/ml and avoided drinking the highest concentration of nicotine solution tested. It is possible that the decrease in drinking is due to negative consequences of hyperactivation of the autonomic nervous system, leading to gastric distress or nausea. However, we observed no significant differences in body weight (Figure 6D), micturition, and digestion (Figure S4) before and during the nicotine consumption experiments. As an independent measure of the effects of nicotine in Tabac mice, CPA assays were performed. Because conditioning to nicotine is both concentration and strain dependent (O’Dell and Khroyan, 2009) we measured CPA in WT C57BL/6 littermates at 0.5 mg nicotine/kg body weight. Under these conditions, we observed neither a preference for nor aversion to nicotine. In contrast, strong CPA selleck chemicals llc to nicotine was observed in Tabac mice (Figure 6E). These data both confirm the conclusions of the nicotine consumption assays, and demonstrate that negative reward learning associated with nicotine is strongly increased in Tabac mice. We conclude that overexpression PTPRJ of the β4 subunit

in vivo leads to an increase in functional α3β4∗ receptors, resulting in a higher sensitivity to the aversive properties of nicotine. The observations that the α5 D397N variant reduces α3β4α5 nicotine-evoked currents in oocytes (Figure 1), and that the MHb-IPN tract contains a high density of native α5 nAChR subunits in combination with α3β4 subunits (Figure 3), suggested that the enhanced nicotine aversion evident in Tabac mice could be reversed by expression of the α5 variant in the MHb. To test this hypothesis we employed lentiviral-mediated transduction to express the α5 D397N in MHb neurons of Tabac mice. We injected bilaterally either control lentivirus (LV-PC) or the LV-α5 D397N (LV-α5N) viruses in Tabac mice. As shown in Figure 7B, immunostaining for the mCherry reporter of LV-α5N expression or direct fluorescence derived from the control lentivirus demonstrated that the lentiviral-transduced area corresponds with that occupied by α3β4∗/eGFP-labeled neurons in the MHb of Tabac mice.

2 Increased PA participation selleck screening library among international students may also provide opportunities to increase intercultural communication and understanding, and help reduce instances of racism and other forms of discrimination, exclusion, and resentment.4 Given the innumerable cognitive, health, and social benefits associated with PA for all people,5 identifying factors that influence Chinese international students’

PA participation is eminently important. Understanding the PA experiences of Chinese international college students is also distinct due to their unique backgrounds. For example, traditional Chinese health beliefs value harmony with nature, which may diminish one’s desire to partake in un-natural actions, such as PA, to change their health status.6 In addition, since the majority of colleges and universities in China lack comprehensive

physical education classes and equipment,7 Chinese students may lack the skills needed to use the exercise facilities that are available on American college and university campuses. Additionally, and similar to other immigrants, Chinese international students may encounter significant obstacles to their PA participation on the basis of their gender, ethnicity, and social class, among other factors.4 As a result, it is important to explore the specific factors influencing their PA participation. Ultimately such CAL101 knowledge can help in the design and delivery of culturally acceptable and maximally effective PA intervention programs. The youth Moxisylyte physical activity promotion (YPAP) model offers a potentially useful framework for understanding Chinese international college students’ PA behavior.5 It was developed from the PRECEDE-PROCEED model, which proposes that in order to design interventions to change health behavior, steps including social diagnosis, epidemiological diagnosis, behavior

and environmental diagnosis, and educational and organizational diagnosis need to be followed.8 Within the educational and organizational diagnosis phase, factors that influence the specific health behavior should be identified, including the predisposing, reinforcing, and enabling factors. The YPAP builds off of the PRECEDE-PROCEED model and adds further specificity. In accordance with the PRECEDE-PROCEED model, the YPAP model explores the mechanisms of youth PA behavior by identifying predisposing, enabling, and reinforcing factors. Predisposing factors include two parts, “Am I able?” and “Is it worth it?” (simplified as able and worth, respectively, in the following). The able construct relates to self-perceptions of physical ability, including self-efficacy and perceived competence. The worth construct addresses the value (i.e., benefits and costs) placed on expected outcomes associated with PA, including attitude, belief, enjoyment, and knowledge.

They injected small (1–2 μL) amounts of OGB-1 to stain a column-like region in cortex 0.5 mm in diameter and to stain small portions of visual and somatosensory thalamic nuclei. In isoflurane-anesthetized mice, the fluorescent calcium signals showed large spontaneous Selumetinib manufacturer population transients in the primary visual cortex recurring with a frequency of 8–30 per minute, most likely reflecting the depolarization and spike firing that characterize the up-states of the slow oscillation (Chauvette et al., 2010; Contreras and Steriade, 1995; Sanchez-Vives and McCormick,

2000; Steriade et al., 1993; Wester and Contreras, 2012). Visual stimulation with brief light flashes triggered population calcium transients that were of the same amplitude and duration as those occurring spontaneously, suggesting that, as recorded with voltage sensitive dyes in response to whisker stimulation (Civillico

and Contreras, 2012; Ferezou et al., 2007), up-states are generated within cortical circuits independently of their triggering mechanism. Work in vitro (Sanchez-Vives and McCormick, 2000) and in vivo (Chauvette et al., 2010) provided evidence that the slow oscillation originates in cortical layer 5 (L5). To demonstrate causality between activation of L5 and the generation of population activity in cortex, the authors used transgenic Thy-1-ChR2 mice that express ChR2 in L5 neurons. They demonstrate that optogenetic stimulation of L5 with brief (50 ms) pulses of blue light generate population calcium transients with similar amplitude and duration selleck as those triggered by visual stimulation or occurring spontaneously. The authors then asked how many L5 neurons are necessary to initiate a population calcium transient? In order to activate small populations of L5 primary visual cortical neurons, through they expressed ChR2 exclusively in a small

region of L5 using viral transduction. Using confocal imaging, they showed that transfection was indeed limited to the targeted layer and was confined over an area of about 1 mm in diameter. The authors then activated a region of about half that diameter (0.5 mm) with blue light to stimulate approximately 200 transfected neurons. Under these conditions, 200 ms pulses of light triggered all-or-none calcium transients in more than 70% of cases. By titrating the number of transfected cells using small amounts of viral solution, they show that activation of as few as 60 L5 neurons is sufficient to initiate a calcium transient. This result demonstrates the enormous amplification power of cortical recurrent networks in L5. To test whether supragranular layers are also capable of generating population calcium transients, the authors targeted small viral injections to layer 2/3 (L2/3). In stark contrast with L5, optogenetic stimulation of L2/3 did not generate calcium waves even at the highest laser intensity.