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Genetically encoded fluorescent voltage sensors have long been desired to report voltage in defined subsets of neurons with optical readout.
Here, we illustrate an alternative approach to drive Cre expression specifically in defined subsets of CNS projection neurons, so as to map both direct and indirect presynaptic inputs to these cells.
We recently showed that MLV-derived vectors integrate preferentially in hot spots around cell-specific genes, enriched in defined subsets of transcription factor binding sites (TFBSs), and suggested that MLV pre-integration complexes (PICs) are tethered to transcriptionally active regulatory regions engaged by basal components of the RNA polymerase II (Pol II) transcriptional machinery [15], [16].
To achieve sparse cell-type specific Cre-mediated recombination, we chose a strategy in which an internal ribosome entry site (IRES) and a CreER coding region were inserted into the 3' untranslated region (UTR) of three genes that are expressed in defined subsets of neurons and one gene that is expressed in neurons and a subset of glia (Figure 1).
While this last challenge is being partially addressed by the creation of BAC transgenic lines that express GFP or Cre in defined subsets of cells [e.g. refs 49], [50], the challenge of visualizing individual neuronal morphologies can only be met by a method that generates a relatively sparse collection of labeled neurons.
Breast cancer risk in defined subsets was assessed using standardized incidence ratios (SIRs), relative to the Iowa Surveillance, Epidemiology, and End Results registry.
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The identification of molecular markers, such as p53, thymidylate synthase, K-ras, and others, may provide an important tool for medical oncologists in defining subsets of patients with gastrointestinal cancers more suitable to benefit from chemotherapy or from experimental therapies.
In this work, however, we did not utilize OS-EM due to the difficulties in defining subsets from 19 projections.
These heterogeneities suggest an important role for the AIS in defining subsets of motor neurons with different spiking properties.
Our finding that NPY positively correlated with perceived stress, anger, depression, negative thoughts and maladaptive coping in patients with CFS suggests that NPY will be useful in defining subsets of patients for clinical trials and as a measure of therapeutic effects.
This unusual form of action potential integration and persistent neural activity (barrage firing) has been observed in genetically defined subsets of interneurons of several brain areas, both in vitro and in vivo22,23,24,25,26.
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