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Related Concept Videos

Phenotypic Profiling of Human Stem Cell-Derived Midbrain Dopaminergic Neurons09:21

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This protocol describes the cell culturing of human midbrain dopaminergic neurons, followed by immunological staining and the generation of neuronal phenotypic profiles from acquired microscopic high-content images allowing the identification of phenotypic variations due to genetic or chemical...
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Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression10:54

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In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using...
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The causes of degeneration of midbrain dopaminergic neurons during Parkinson’s disease are not fully understood. Cellular culture systems provide an essential tool for study of the neurophysiological properties of these neurons. Here we present an optimized protocol, which can be utilized for in vitro modeling of...
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This protocol describes two different environmental manipulations and a concurrent brain infusion protocol to study environmentally-induced brain changes underlying adaptive behavior and brain repair in adult...
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Dopaminergic neurons play a vital regulatory role in the brain. Their loss is associated with Parkinson's disease. In this video, we show how to generate primary cultures of central dopaminergic neurons from embryonic mouse mesencephalon. Such cultures are useful to study the extreme vulnerability of these neurons to various...
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Dopamine is distinctly regulated in the midbrain nuclei, which contain the cell bodies and dendrites of the dopamine neurons. Here we describe a dissection and sample-handling approach to maximize results, and thus conclusions and insights, on dopamine regulation in the midbrain nuclei of the substantia nigra (SN) and ventral tegmental area (VTA) in...
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Related Experiment Video

Updated: Jan 8, 2026

Environmental Modulations of the Number of Midbrain Dopamine Neurons in Adult Mice
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Sequence determinants of promoter activity.

P Youderian, S Bouvier, M M Susskind

    Cell
    |October 1, 1982
    PubMed
    Summary

    Mutations in the bacteriophage P22 antirepressor (ant) promoter reduce its activity. These promoter-down mutations reveal critical regions and highlight conserved sequences as key determinants of promoter strength.

    Area of Science:

    • Bacteriophage genetics
    • Molecular biology
    • Promoter regulation

    Background:

    • The P22 bacteriophage antirepressor (ant) promoter (Pant) normally produces high levels of antirepressor.
    • Overproduction of antirepressor inhibits progeny phage production during lytic infection.
    • Pseudorevertants of P22 arc-amber phage often possess mutations reducing Pant activity.

    Purpose of the Study:

    • To identify and characterize mutations affecting the P22 Pant activity.
    • To define regions critical for Pant function.
    • To understand the relationship between promoter sequence homology and promoter strength.

    Main Methods:

    • DNA sequence analysis of 72 independent Pant promoter-down mutations.
    • Comparison of mutated Pant sequences with the consensus promoter sequence.

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  • Assessment of mutation effects on promoter activity.

    • Main Results:

    • Over 25 distinct alterations were identified in the Pant promoter.
    • Mutations clustered in two regions critical for promoter activity.
    • Most mutations reduced homology with the consensus promoter sequence.
    • Substitutions at the same site generally had similar effects on promoter strength.

    Conclusions:

    • Conserved features in promoter sequences are critical determinants of promoter strength.
    • The identified mutations provide insights into Pant promoter function and regulation.
    • Understanding promoter strength is crucial for bacteriophage gene expression and replication.