A biotin-containing compound N-(2-aminoethyl)biotinamide for intracellular labeling and neuronal tracing studies

Comparison with biocytin

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Abstract

The hydrochloride salt of a new, small molecular weight (M.W. = 286) biotin-containing compound referred to as biotinamide (N-(2-aminoethyl)biotinamide) was compared with biocytin (M.W. = 372) for its use in intracellular labeling of neurons and in neuronal tracing experiments using avidin conjugates for histochemical detection. The DC resistance and current passing ability of electrodes filled with 1-2 M potassium chloride, potassium acetate or potassium methylsulfate and containing 1-4% of these compounds were compared. Although differences were observed due to the electrolyte, with KC1 electrodes being the least resistant, no electrode differences could be attributed to the concentration or type of tracer. However, whereas biocytin could be electrophoresed with either positive or negative current with roughly similar facility, biotinamide was selectively ejected with positive current. This would be beneficial to electrophysiologists using hyperpolarizing current to stabilize the membrane potential of neurons prior to recording. In addition, biotinamide-HCl could be dissolved at concentrations of 2-4% in either 1 or 2 M salt without precipitation, whereas biocytin precipitated in some of these solutions. Both compounds were equally useful for neuronal tracing experiments with survival times of 2 days, but labeling was much weaker with longer survival times. There was also little difference in the ability to histochemically localize these compounds using avidin conjugates, including avidin-biotin-horseradish peroxidase complex. In conclusion, biotinamide shares many of the useful features of biocytin, but can be selectively electrophoresed with positive current and can be dissolved at higher concentrations with little detriment in the electrical properities of the recording electrode.

Original languageEnglish (US)
Pages (from-to)141-150
Number of pages10
JournalJournal of Neuroscience Methods
Volume37
Issue number2
DOIs
StatePublished - Jan 1 1991

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Biotin
Electrodes
Avidin
Potassium Acetate
Salts
Neurons
Potassium Chloride
Membrane Potentials
Electrolytes
Potassium
Molecular Weight
biocytin
biotinamide
neurobiotin

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

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title = "A biotin-containing compound N-(2-aminoethyl)biotinamide for intracellular labeling and neuronal tracing studies: Comparison with biocytin",
abstract = "The hydrochloride salt of a new, small molecular weight (M.W. = 286) biotin-containing compound referred to as biotinamide (N-(2-aminoethyl)biotinamide) was compared with biocytin (M.W. = 372) for its use in intracellular labeling of neurons and in neuronal tracing experiments using avidin conjugates for histochemical detection. The DC resistance and current passing ability of electrodes filled with 1-2 M potassium chloride, potassium acetate or potassium methylsulfate and containing 1-4{\%} of these compounds were compared. Although differences were observed due to the electrolyte, with KC1 electrodes being the least resistant, no electrode differences could be attributed to the concentration or type of tracer. However, whereas biocytin could be electrophoresed with either positive or negative current with roughly similar facility, biotinamide was selectively ejected with positive current. This would be beneficial to electrophysiologists using hyperpolarizing current to stabilize the membrane potential of neurons prior to recording. In addition, biotinamide-HCl could be dissolved at concentrations of 2-4{\%} in either 1 or 2 M salt without precipitation, whereas biocytin precipitated in some of these solutions. Both compounds were equally useful for neuronal tracing experiments with survival times of 2 days, but labeling was much weaker with longer survival times. There was also little difference in the ability to histochemically localize these compounds using avidin conjugates, including avidin-biotin-horseradish peroxidase complex. In conclusion, biotinamide shares many of the useful features of biocytin, but can be selectively electrophoresed with positive current and can be dissolved at higher concentrations with little detriment in the electrical properities of the recording electrode.",
author = "Hitoshi Kita and William Armstrong",
year = "1991",
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T1 - A biotin-containing compound N-(2-aminoethyl)biotinamide for intracellular labeling and neuronal tracing studies

T2 - Comparison with biocytin

AU - Kita, Hitoshi

AU - Armstrong, William

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Y1 - 1991/1/1

N2 - The hydrochloride salt of a new, small molecular weight (M.W. = 286) biotin-containing compound referred to as biotinamide (N-(2-aminoethyl)biotinamide) was compared with biocytin (M.W. = 372) for its use in intracellular labeling of neurons and in neuronal tracing experiments using avidin conjugates for histochemical detection. The DC resistance and current passing ability of electrodes filled with 1-2 M potassium chloride, potassium acetate or potassium methylsulfate and containing 1-4% of these compounds were compared. Although differences were observed due to the electrolyte, with KC1 electrodes being the least resistant, no electrode differences could be attributed to the concentration or type of tracer. However, whereas biocytin could be electrophoresed with either positive or negative current with roughly similar facility, biotinamide was selectively ejected with positive current. This would be beneficial to electrophysiologists using hyperpolarizing current to stabilize the membrane potential of neurons prior to recording. In addition, biotinamide-HCl could be dissolved at concentrations of 2-4% in either 1 or 2 M salt without precipitation, whereas biocytin precipitated in some of these solutions. Both compounds were equally useful for neuronal tracing experiments with survival times of 2 days, but labeling was much weaker with longer survival times. There was also little difference in the ability to histochemically localize these compounds using avidin conjugates, including avidin-biotin-horseradish peroxidase complex. In conclusion, biotinamide shares many of the useful features of biocytin, but can be selectively electrophoresed with positive current and can be dissolved at higher concentrations with little detriment in the electrical properities of the recording electrode.

AB - The hydrochloride salt of a new, small molecular weight (M.W. = 286) biotin-containing compound referred to as biotinamide (N-(2-aminoethyl)biotinamide) was compared with biocytin (M.W. = 372) for its use in intracellular labeling of neurons and in neuronal tracing experiments using avidin conjugates for histochemical detection. The DC resistance and current passing ability of electrodes filled with 1-2 M potassium chloride, potassium acetate or potassium methylsulfate and containing 1-4% of these compounds were compared. Although differences were observed due to the electrolyte, with KC1 electrodes being the least resistant, no electrode differences could be attributed to the concentration or type of tracer. However, whereas biocytin could be electrophoresed with either positive or negative current with roughly similar facility, biotinamide was selectively ejected with positive current. This would be beneficial to electrophysiologists using hyperpolarizing current to stabilize the membrane potential of neurons prior to recording. In addition, biotinamide-HCl could be dissolved at concentrations of 2-4% in either 1 or 2 M salt without precipitation, whereas biocytin precipitated in some of these solutions. Both compounds were equally useful for neuronal tracing experiments with survival times of 2 days, but labeling was much weaker with longer survival times. There was also little difference in the ability to histochemically localize these compounds using avidin conjugates, including avidin-biotin-horseradish peroxidase complex. In conclusion, biotinamide shares many of the useful features of biocytin, but can be selectively electrophoresed with positive current and can be dissolved at higher concentrations with little detriment in the electrical properities of the recording electrode.

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