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Noradrenergic terminal density varies among different groups of hypoglossal premotor neurons

Boyle, CE;Parkar, A;Barror, A;Kubin, L;
Product: Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

In obstructive sleep apnea (OSA) patients, contraction of the muscles of the tongue is needed to protect the upper airway from collapse. During wakefulness, norepinephrine directly excites motoneurons that innervate the tongue and other upper airway muscles but its excitatory effects decline during sleep, thus contributing to OSA. In addition to motoneurons, NE may regulate activity in premotor pathways but little is known about these upstream effects. To start filling this void, we injected a retrograde tracer (beta-subunit of cholera toxin-CTb; 5-10 nl, 1%) into the hypoglossal (XII) motor nucleus in 7 rats. We then used dual immunohistochemistry and brightfield microscopy to count dopamine beta-hydroxylase (DBH)-positive axon terminals closely apposed to CTb cells located in five anatomically distinct XII premotor regions. In different premotor groups, we found on the average 2.2-4.3 closely apposed DBH terminals per cell, with ˜60% more terminals on XII premotor neurons located in the ventrolateral pontine parabrachial region and ventral medullary gigantocellular region than on XII premotor cells of the rostral or caudal intermediate medullary reticular regions. This difference suggests stronger control by norepinephrine of the interneurons that mediate complex behavioral effects than of those mediating reflexes or respiratory drive to XII motoneurons. Copyright © 2019 Elsevier B.V. All rights reserved.

PubMed ID: 31128245
359235922019-06-172019-06-1714:32:1914:32:192019-07-052019-07-0515:14:3815:14:38Boyle, CE;Parkar, A;Barror, A;Kubin, L;Boyle, CE;Parkar, A;Barror, A;Kubin, L;20192019Noradrenergic terminal density varies among different groups of hypoglossal premotor neuronsNoradrenergic terminal density varies among different groups of hypoglossal premotor neuronsJournal Of Chemical NeuroanatomyJournal Of Chemical Neuroanatomy1016511016511001003112824531128245

In obstructive sleep apnea (OSA) patients, contraction of the muscles of the tongue is needed to protect the upper airway from collapse. During wakefulness, norepinephrine directly excites motoneurons that innervate the tongue and other upper airway muscles but its excitatory effects decline during sleep, thus contributing to OSA. In addition to motoneurons, NE may regulate activity in premotor pathways but little is known about these upstream effects. To start filling this void, we injected a retrograde tracer (beta-subunit of cholera toxin-CTb; 5-10 nl, 1%) into the hypoglossal (XII) motor nucleus in 7 rats. We then used dual immunohistochemistry and brightfield microscopy to count dopamine beta-hydroxylase (DBH)-positive axon terminals closely apposed to CTb cells located in five anatomically distinct XII premotor regions. In different premotor groups, we found on the average 2.2-4.3 closely apposed DBH terminals per cell, with ˜60% more terminals on XII premotor neurons located in the ventrolateral pontine parabrachial region and ventral medullary gigantocellular region than on XII premotor cells of the rostral or caudal intermediate medullary reticular regions. This difference suggests stronger control by norepinephrine of the interneurons that mediate complex behavioral effects than of those mediating reflexes or respiratory drive to XII motoneurons. Copyright © 2019 Elsevier B.V. All rights reserved.

In obstructive sleep apnea (OSA) patients, contraction of the muscles of the tongue is needed to protect the upper airway from collapse. During wakefulness, norepinephrine directly excites motoneurons that innervate the tongue and other upper airway muscles but its excitatory effects decline during sleep, thus contributing to OSA. In addition to motoneurons, NE may regulate activity in premotor pathways but little is known about these upstream effects. To start filling this void, we injected a retrograde tracer (beta-subunit of cholera toxin-CTb; 5-10 nl, 1%) into the hypoglossal (XII) motor nucleus in 7 rats. We then used dual immunohistochemistry and brightfield microscopy to count dopamine beta-hydroxylase (DBH)-positive axon terminals closely apposed to CTb cells located in five anatomically distinct XII premotor regions. In different premotor groups, we found on the average 2.2-4.3 closely apposed DBH terminals per cell, with ˜60% more terminals on XII premotor neurons located in the ventrolateral pontine parabrachial region and ventral medullary gigantocellular region than on XII premotor cells of the rostral or caudal intermediate medullary reticular regions. This difference suggests stronger control by norepinephrine of the interneurons that mediate complex behavioral effects than of those mediating reflexes or respiratory drive to XII motoneurons. Copyright © 2019 Elsevier B.V. All rights reserved.

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Retrograde tracer injections:

Injections of cholera toxin beta sub-unit (CTb; List Biological Laboratories, Campbell, CA) were used to retrogradely label premotor cells with axonal projections to the XII motor nucleus, ...

 

Author did not specify which CTB was utilized.  List Labs has provided Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae) and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).  The former has been discontinued.  The latter may be substituted for the former, if the researcher takes into account the difference in the buffer in which each product is provided and the amount of protein per vial.  When Product #103B was reconstituted with 200 μl of water, it contained 1 mg of protein in 0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA, 0.003 M NaN3, pH 7.5.  When Product #104 is reconstituted with 0.25 ml water, it contains 0.5 mg of Cholera Toxin B Subunit (choleragenoid) in 0.01 M Sodium Phosphate at pH 7.5.

Retrograde tracer injections:

Injections of cholera toxin beta sub-unit (CTb; List Biological Laboratories, Campbell, CA) were used to retrogradely label premotor cells with axonal projections to the XII motor nucleus, ...

 

Author did not specify which CTB was utilized.  List Labs has provided Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae) and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).  The former has been discontinued.  The latter may be substituted for the former, if the researcher takes into account the difference in the buffer in which each product is provided and the amount of protein per vial.  When Product #103B was reconstituted with 200 μl of water, it contained 1 mg of protein in 0.05 M Tris, 0.2 M NaCl, 0.001 M Na2EDTA, 0.003 M NaN3, pH 7.5.  When Product #104 is reconstituted with 0.25 ml water, it contains 0.5 mg of Cholera Toxin B Subunit (choleragenoid) in 0.01 M Sodium Phosphate at pH 7.5.

https://www.sciencedirect.com/science/article/pii/S0891061819300341https://www.sciencedirect.com/science/article/pii/S08910618193003412019-05-222019-05-2210.1016/j.jchemneu.2019.10165110.1016/j.jchemneu.2019.101651Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low SaltCholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Saltlkubin@vet.upenn.edulkubin@vet.upenn.eduActivity;Axon;Biological;Cell;Cholera;Complex;Control;CTB;Immunohistochemistry;Label;List;List Biological;Nucleus;Positive;Regulate;Respiratory;Retrograde;Subunit;Terminal;Toxin;Ventral;Journal Of Chemical NeuroanatomyActivity;Axon;Biological;Cell;Cholera;Complex;Control;CTB;Immunohistochemistry;Label;List;List Biological;Nucleus;Positive;Regulate;Respiratory;Retrograde;Subunit;Terminal;Toxin;Ventral;Journal Of Chemical Neuroanatomy104104noradrenergic-terminal-density-varies-amongnoradrenergic-terminal-density-varies-among