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Science 338 (6106): 540-543

Copyright © 2012 by the American Association for the Advancement of Science

Oxytocin/Vasopressin-Related Peptides Have an Ancient Role in Reproductive Behavior

Jennifer L. Garrison, Evan Z. Macosko, Samantha Bernstein, Navin Pokala, Dirk R. Albrecht, and Cornelia I. Bargmann

Howard Hughes Medical Institute, Lulu and Anthony Wang Laboratory of Neural Circuits and Behavior, The Rockefeller University, New York, NY 10065, USA.


Figure 1
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Fig. 1. ntc-1 encodes an oxytocin/vasopressin-related peptide that signals through receptors encoded by ntr-1 and ntr-2. (A) Gene models with deletions in mutant alleles indicated by horizontal black bars (www.wormbase.org). (B and C) Domain structure and amino acid sequence of vasopressin (AVP), oxytocin (OXT), and nematocin (NTC-1) precursors and peptides. (D and E) Mean calcium responses and dose-response curves of fura-2/AM-loaded HEK293T cells transfected with ntr-1 and Gα15 and exposed to synthetic NTC-1 peptide. (F) Forskolin-stimulated cAMP production in NTC-1–treated HEK293T cells transfected with ntr-1, ntr-2, and Gα15. ***P < 0.0001. Error bars in all figures indicate SEM.

 

Figure 2
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Fig. 2. ntc-1, ntr-1, and ntr-2 expression patterns. P, gene promoter fragment used. The upper panels show expression in adult hermaphrodite heads; the lower panels show male [(B), (D), and (F)] and hermaphrodite (H) tails. (A to F) Expression of ntc-1, ntr-1, and ntr-2 reporter genes (table S1). (G and H) Immunostaining of AFD, NSM, and DVA neurons with antisera to nematocin. White arrows indicate expression in specified neurons. Asterisks mark autofluorescence; scale bars represent 10 μm.

 

Figure 3
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Fig. 3. Nematocin-deficient males exhibit mating defects. (A) Steps in male mating behavior. The black arrow notes the direction of male movement. (B) Reproductive efficiencies of wild-type (WT, gray bars) and mutant males (black bars) in long-term single-pair mating crosses (no. of cross-progeny/no. of total progeny). (C) Percentage of males that successfully transferred sperm within 5 min of first tail contact with a hermaphrodite. (D) Mean number of hermaphrodites that a male’s tail contacted before he initiated mating. (E) Mean total number of turns a male executed around the hermaphrodite’s body before locating the vulva. (F) Summary of the behavioral transitions in ethograms of male mating behavior (figs. S8 and S9); the arrow width indicates the probability of behavioral transitions. (G) Male mate search behavior, measured as the rate at which a single male leaves food in the absence of hermaphrodites (15). *P < 0.01, **P < 0.001, ***P < 0.0001.

 

Figure 4
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Fig. 4. Selective inactivation of ntc-1 in DVA affects individual steps of male mating behavior. (A) Cre/Lox strategy to selectively inactivate ntc-1 in DVA. (Inset) Tail of larval stage 4 male expressing mCherry and not GFP in DVA, indicating successful DVA-specific recombination. Scale bar, 10 μm. (B to F) Behavioral phenotypes of males with ntc-1 inactivated in DVA [(C) to (F)] or after DVA laser ablation (B). (C) to (E) Behaviors as in Fig. 3. (F) Vulva location ability of males (percentage of positive vulva detections/total vulva encounters). *P < 0.01, **P < 0.001, ***P < 0.0001.

 


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