DOI: 10.5061/dryad.12jm63z6t
title: Reproductive experience drives changes in behavior and physiology in male California mice (Peromyscus californicus)

description: In biparental mammals, male behavior and reproductive physiology may be
 shaped by mating, siring, and caregiving, but their individual
 contributions are less understood. This dataset includes raw behavioral,
 physiological, and caregiving measures from two experiments
 in Peromyscus californicus males. In Experiment 1, we compared
 virgins, nonfathers (mated with sterile females), and experienced fathers
 on anxiety-like behavior (elevated plus maze), recognition memory (novel
 object recognition), testes weight, and sperm count. In Experiment 2, we
 compared nonfathers, pup-sensitized nonfathers (exposed to unrelated
 pups), and first-time fathers on the same behavioral and reproductive
 measures, as well as pup-directed behaviors including grooming, latency to
 approach, and nest building. Experienced fathers in Experiment 1 showed
 enhanced recognition memory, reduced anxiety-like behavior, and greater
 reproductive investment compared to virgins and nonfathers. In Experiment
 2, only testes weight differed among groups—first-time fathers had larger
 testes than nonfathers—suggesting that reproductive investment responds
 more rapidly to mating and siring than behavioral traits do to brief
 caregiving. Pup-sensitized males and fathers differed in caregiving
 strategies: the former groomed pups more, while the latter invested more
 in nest building. The dataset includes CSV files of all raw data and R
 scripts for linear and mixed effects models.
description: ## **Overview** This repository contains data and analysis scripts related
 to our behavioral and reproductive trait studies. The dataset includes raw
 data from various experiments and corresponding R scripts that perform
 linear models and linear mixed effects models on the data.   ## **Files**
 ### **Elevated Plus Maze (EPM) Data and Scripts** 1. **EPM_Exp_1.csv** *
 Description: Contains the raw data from the Experiment 1 elevated plus
 maze test. * Variables: 1. Subject: A unique identifier assigned to each
 animal tested. 2. Treatment: Experimental group designation. 3.
 Distance_moved: Total distance the subject moved (in cm) during the EPM
 trial. 4. NUM_Closed_arm (Number of Closed Arm Entries): The number of
 times the subject entered either of the two closed arms. 5. DUR_Closed_arm
 (Duration in Closed Arms): Total time (in seconds) spent in the closed
 arms during the trial. 6. NUM_Open_arm (Number of Open Arm Entries): The
 number of times the subject entered either of the two open arms. 7.
 DUR_Open_arm (Duration in Open Arms): Total time (in seconds) spent in the
 open arms during the trial. 8. Prcnt_open_arm (Percent Time in Open Arms):
 The percentage of the total trial time spent in the open arms. 9.
 Age_at_EPM: The age of the subject in days at the time of EPM testing.
 ‘NA’ indicates age was unknown. 2. **EPM_Exp_2.csv** * Description:
 Contains the raw data from the Experiment 2 elevated plus maze test. *
 Variables: 1. Subject: A unique identifier assigned to each animal tested.
 2. Treatment: Experimental group designation. ‘PSNF’ indicates
 pup-sensitized nonfathers. 3. Trial: Indicates test number on EthoVision
 software. 4. Male_age_at_testing: The age of the subject in days at the
 time of the EPM testing. 5. NUM_Closed_arms (Number of Closed Arm
 Entries): The number of times the subject entered either of the two closed
 arms. 6. DUR_Closed_arms (Duration in Closed Arms): Total time (in
 seconds) spent in the closed arms during the trial. 7. LAT_Closed_arms
 (Latency to Enter Closed Arms): Time (in seconds) from the start of the
 trial to the subject's first entry into a closed arm. 8.
 NUM_Opened_arms (Number of Open Arm Entries): The number of times the
 subject entered either of the two open arms. 9. DUR_Open_arms (Duration in
 Open Arms): Total time (in seconds) spent in the open arms during the
 trial. 10. LAT_Open_arms (Latency to Enter Open Arms): Time (in seconds)
 from trial start to the subject's first entry into an open arm. 11.
 Total_time: Total duration of the trial in seconds. 12. Distance_moved:
 Total distance the subject moved (in cm) during the EPM trial. 13.
 Percent_open_arms_w_center (% Time in Open Arms Including Center): The
 percentage of the trial spent in the open arms including any time the
 animal spent in the center platform of the maze (which connects the arms).
 14. Percent_open_arms_wo_center (% Time in Open Arms Excluding Center):
 The percentage of time spent in the open arms excluding time spent in the
 center zone. 3. **EPM_LM_G.R** * Description: R script that contains the
 code used to perform linear models on the elevated plus maze data for
 Experiments 1 and 2.   ### **Novel Object Recognition (NOR) Data and
 Scripts** 1. **NOR_Exp_1.csv** * Description: Contains the raw data from
 the Experiment 1 novel object recognition test. * Variables: 1. Subject: A
 unique identifier assigned to each animal tested. 2. Treatment:
 Experimental group designation. 3. Age_at_NOR: The age of the subject in
 days at the time of NOR testing. ‘NA’ indicates age was unknown. 4.
 Marble_time (Time with Familiar Object): The total amount of time (in
 seconds) the subject spent interacting with the familiar marble. 5.
 Block_time (Time with Novel Object): The total amount of time (in seconds)
 the subject spent interacting with the novel block. 6. Prcnt_novel_object
 (Percent Time with Novel Object): The percentage of total object
 interaction time spent investigating the novel object. Calculated as
 (Block_time) / (Marble_time + Block_time) ​× 100. 7. Difference_score: A
 measure of preference calculated as Block_time minus Marble_time. 2.
 **NOR_Exp_2.csv** * Description: Contains the raw data from the Experiment
 2 novel object recognition test. * Variables: 1. Treatment: Experimental
 group designation. ‘PSNF’ indicates pup-sensitized nonfathers. 2. Subject:
 A unique identifier assigned to each animal tested. 3. Difference_score: A
 measure of preference calculated as Block_time minus Marble_time. 4.
 Percent_novel_object (Percent Time with Novel Object): The percentage of
 total object interaction time spent investigating the novel object.
 Calculated as (Block_time) / (Marble_time + Block_time) ​× 100. 5.
 Block_time (Time with Novel Object): The total amount of time (in seconds)
 the subject spent interacting with the novel block. 6. Marble_time (Time
 with Familiar Object): The total amount of time (in seconds) the subject
 spent interacting with the familiar marble. 7. Male_age_at_testing: The
 age of the subject in days at the time of NOR testing. 8.
 Female_mating_history: The number of males to which a female was paired.
 3. **NOR_LM_G.R** * Description: R script that contains the code used to
 perform linear models on the novel object recognition data for Experiments
 1 and 2. ### **Reproductive Trait Data and Scripts** 1.
 **Testes_weight_Sperm_count_Exp_1.csv** * Description: Contains the raw
 average testes weight and sperm count data from Experiment 1. * Variables:
 1. ID: A unique identifier assigned to each individual subject. 2.
 Treatment: Experimental group designation. ‘NF’ indicates nonfathers. 3.
 Age: The age of the subject in days. 4. BW (Body Weight): The body weight
 of the subject at the time of dissection, measured in milligrams. 5.
 Relative_testes_weight: Mean testes  weight (in mg) normalized to body
 weight. 6. Sperm_count: Number of sperm collected from the epididymis. 7.
 Mean_testes_weight: The average weight (in mg) of a single testis
 calculated by dividing the combined weight of both testes by two. 2.
 **Sperm_count_Exp_2.csv** * Description: Contains the raw sperm count data
 from Experiment 2. * Variables: 1. ID: A unique identifier assigned to
 each individual subject. 2. Treatment: Experimental group designation.
 ‘NF’ indicates nonfathers and ‘PSNF’ indicates pup-sensitized nonfathers.
 3. BW (Body Weight): The body weight of the subject at the time of
 dissection, measured in milligrams. 4. Age: The age of the subject in
 days. 5. Sperm_Count: Number of sperm collected from the epididymis. Cells
 are left empty if we were unable to collect sperm from subject. 3.
 **Testes_weight_Exp_2.csv** * Description: Contains the raw average testes
 weight data from Experiment 2. * Variables: 1. ID: A unique identifier
 assigned to each individual subject. 2. Treatment: Experimental group
 designation. ‘NF’ indicates nonfathers and ‘PSNF’ indicates pup-sensitized
 nonfathers. 3. BW (Body Weight): The body weight of the subject at the
 time of dissection, measured in milligrams. 4. Age: The age of the subject
 in days. 5. Mean_testes_weight: The average weight of a single testis
 calculated by dividing the combined weight of both testes by two. 4.
 **Testes_weight_Sperm_count_Exp_2.csv** * Description: Contains the raw
 average testes weight and sperm count data from Experiment 2. * Variables:
 1. ID: A unique identifier assigned to each individual subject. 2.
 Treatment: Experimental group designation. ‘NF’ indicates nonfathers and
 ‘PSNF’ indicates pup-sensitized nonfathers. 3. Sperm_Count: Number of
 sperm collected from the epididymis. Cells are left empty if we were
 unable to collect sperm from subject. 4. Mean_testes_weight: The average
 weight (in mg) of a single testis calculated by dividing the combined
 weight of both testes by two. 5. **Repro_LM_G.R** * Description: R script
 that contains the code used to perform linear models on the reproduction
 data for Experiments 1 and 2.   ### **Pup-Exposure and Caregiving Behavior
 Data and Scripts** 1. **Pup_exposure.csv** * Description: Contains the raw
 data on pup-exposure behavior. * Variables: 1. Subject: A unique
 identifier assigned to each animal tested. 2. Treatment: Experimental
 group designation. 3. PND (Postnatal Day): The age of the pups used in the
 assay, in days. 4. Male_age_at_testing: The age of the male subject at the
 time of the test, in days. 5. NUM_pups_sired: The number of offspring the
 subject had sired at the time of testing. This value is therefore NA for
 pup-sensitized nonfathers. 6. DUR_Exploration_near: Total time (in
 seconds) spent exploring near the pup enclosure. 7. NUM_Exploration_near:
 Total number of bouts of exploration near the pup enclosure. 8.
 DUR_Exploration_away: Total time (in seconds) spent exploring areas away
 from the pup enclosure. 9. NUM_Exploration_away: Total number of bouts of
 exploration away from the pup enclosure. 10. DUR_Pup_directed: Total time
 (in seconds) engaged in direct pup interactions (e.g., sniffing, licking,
 touching directed at pups). 11. NUM_Pup_directed: Total number of bouts of
 pup-directed interaction. 12. DUR_Latency_pup_directed: Time (in seconds)
 from the start of the test until the male first engaged in a pup-directed
 behavior. 13. NUM_Latency_pup_directed: Number of latency periods scored
 prior to first pup-directed interaction. 14. DUR_Grooming: Total time (in
 seconds) spent auto-grooming. 15. NUM_Grooming: Total number of
 auto-grooming bouts. 16. DUR_Resting: Total time (in seconds) spent
 resting or inactive. 17. NUM_Resting: Number of distinct resting bouts
 during the test. 18. DUR_Jump/backflips: Total time (in seconds) spent
 engaging in jumping or backflipping. All NA because this is a point
 behavior. 19. NUM_Jump/backflips: Number of bouts of jumping or
 backflipping behaviors. 2. **Caregiving.csv** * Description: Contains the
 raw data on caregiving behavior. * Variables: 1. Subject: A unique
 identifier assigned to each animal tested. 2. Treatment: Experimental
 group designation. 3. PND (Postnatal Day): The age of the pups used in the
 assay, in days. 4. Male_age_at_testing: Age of the male (in days) at the
 time of the caregiving assay. 5. Pup_1_time: Total time (in seconds)
 tested with the first pup. 6. Earliest_latency: Latency (in seconds) to
 the first occurrence of any caregiving-related behavior. 7.
 Parental_behavior_latency: Latency (in seconds) to the first instance of
 huddling, grooming, or retrieving. 8. Huddle_latency: Latency (in seconds)
 to first occurrence of huddling behavior. 9. Huddling: Total duration (in
 seconds) spent huddling over the pup. 10. NUM_Huddling: Number of huddling
 bouts. 11. Groom_latency: Latency (in seconds) to first occurrence of
 grooming behavior directed at the pup. 12. Grooming: Total duration (in
 seconds) spent grooming the pup. 13. NUM_Grooming: Number of pup grooming
 bouts. 14. Huddling_grooming: Combined total duration (in seconds) of
 huddling and grooming behaviors. 15. Sniff_latency: Latency (in seconds)
 to first sniff of the pup. 16. Sniffing: Total time (in seconds) spent
 sniffing the pup. 17. NUM_Sniffing: Number of discrete sniffing bouts. 18.
 Retrieve_latency_1: Latency (in seconds) to first pup retrieval. 19.
 Retrieve_latency_1_fixed: Latency (in seconds) to first pup retrieval
 corrected if retrieval did not take place. NA indicates male did not
 retrieve pup.   20. Retrieving_1: Total duration (in seconds) spent
 retrieving the pup. 21. NUM_Retrieving_1: Number of pup retrievals. 22.
 Pup_interaction: Total duration (in seconds) of any pup-directed
 interaction (e.g., sniffing, grooming, huddling, retrieving). 23.
 Nest_building: Total time (in seconds) spent manipulating nesting
 material. 24. NUM_Nest_building: Number of nest-building bouts. 25.
 General_activity: Total time (in seconds) spent in general movement or
 exploration, not directed toward the pup. 26. NUM_General_activity: Number
 of bouts of general (non-caregiving) activity. 27. Resting: Total time (in
 seconds) spent inactive during the trial. 28. General_resting: Combined
 total duration (in seconds) of general activity and resting behaviors. 29.
 Pup_2_Time: Total time (in seconds) tested with the second pup introduced
 later in the assay. 30. Retrieve_latency_2: Latency (in seconds) to
 retrieve the second pup. 31. Retrieve_latency_2_fixed: Latency (in
 seconds) to retrieve the second pup corrected if retrieval did not take
 place. NA indicates male did not retrieve pup.   32.
 NUM_Lack_of_retrieve_2: Count of failure to retrieve pup 2 (e.g., 1 = did
 not retrieve). 33. Investigate_latency_2: Latency (in seconds) to first
 investigation of pup 2. 34. NUM_Lack_of_investigate_2: Count of failure to
 investigate pup 2 (e.g., 1 = did not investigate). 3.
 **Pup_exposure_LM_G.R** * Description: R script that contains the code
 used to perform linear mixed effects models on the pup-exposure data. 4.
 **Caregiving_LM_G.R** * Description: R script that contains the code used
 to perform linear mixed effects models on the caregiving data. ##
 **Usage** 1. **Data Files**: * The raw data files can be loaded into R. 2.
 **Script Files**: * The analysis scripts can be run in an R environment.
 Ensure that you set the working directory to the location of the data
 files or provide the correct path to the data files within the scripts. *
 The scripts will perform the specified modeling on the provided data and
 output results including model summaries.  ## **Contact** For any
 questions or further information, please contact Maria Colt at
 [mcolt@umd.edu](mailto:mcolt@umd.edu).