Dataset: Nematostella ocean acidification experiment: Sperm MMP

Name: Mitochondrial membrane potential (MMP) of male gametes following parental ocean acidification during lab experiments conducted in spring 2022.
Published: 2024-03-24
Keywords: oceans

Cite This Dataset

DOI:10.26008/1912/bco-dmo.920827.1

Laboratory at the University of Pennsylvania

Temporal Extent: 2022-01-26 - 2022-05-02

Project:

Influence of environmental pH variability and thermal sensitivity on the resilience of reef-building corals to acidification stress (Coral Resilience)

Principal Investigator:

Katie Barott (University of Pennsylvania, Penn)

Scientist:

Kristen Brown (University of Pennsylvania, Penn)

Kelsey Speer (University of Pennsylvania, Penn)

Student:

Benjamin Glass (University of Pennsylvania, Penn)

Angela Schmitt (University of Pennsylvania, Penn)

Contact:

Benjamin Glass (University of Pennsylvania, Penn)

BCO-DMO Data Manager:

Karen Soenen (Woods Hole Oceanographic Institution, WHOI BCO-DMO)

Version:

Version Date:

2024-03-24

Restricted:

Validated:

Yes

Current State:

Final no updates expected

Mitochondrial membrane potential (MMP) of male gametes following parental ocean acidification during lab experiments conducted in spring 2022.

Abstract:

Ocean acidification (OA) resulting from anthropogenic CO2 emissions is impairing the reproduction of marine organisms. While parental exposure to OA can protect offspring via carryover effects, this phenomenon is poorly understood in many marine invertebrate taxa. We examined how parental exposure to acidified (pH 7.40) versus ambient (pH 7.72) seawater influenced reproduction and offspring performance across six gametogenic cycles (13 weeks) in the estuarine sea anemone Nematostella vectensis. This dataset pertains to the physiology of gametes released by adults in the OA treatment, specifically the mitochondrial membrane potential (MMP) of sperm.

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Archival Copy

Version Date	Archive	Persistent Identifier	Date Assigned
2024-03-24	Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA)	10.26008/1912/bco-dmo.920827.1	2024-03-24

Description

Data generated as part of a Nematostella ocean acidfication experiment published in Glass et al., 2023. (see Related Publications). Related Zenodo datasets provides further analysis and plotting of the BCO-DMO dataset here. (see Related Dataset).

Methods & Sampling

Nematostella vectensis (Stephenson, 1935) anemones were collected from a salt marsh in Brigantine, New Jersey in the fall of 2020. Females were identified by inducing spawning, and 14 individuals that released eggs were chosen as the genotype pool for this experiment. Each female was then horizontally bisected through the body column using a razor blade, resulting in two genotypically identical individuals that were divided between the two experimental groups (ambient and acidic).

A clonal male population, also originating from the United States Atlantic coast, was obtained from the laboratory of Dr. Katerina Ragkousi (Amherst College) in the spring of 2021. The male population size was increased via bisection, resulting in a total of 20 genetically identical males for the experiment (N=10 per treatment).

All anemones were kept in 12 parts per thousand (ppt) artificial seawater (ASW; Instant Ocean Reef Crystals^Ⓡ reef salt, Spectrum Brands, Blacksburg, VA, USA) at pH 7.7–8.1 and 18°C. The animals were maintained in a dark incubator (Boekel Scientific, Feasterville-Trevose, PA, USA) and fed approximately every other day with Artemia nauplii. The experiment was performed approximately 1–1.5 years after animal collection.

Data Processing Description

Sperm mitochondrial membrane potential (MMP) was measured using the fluorescent dye JC-1 (Thermo Fisher Scientific, Waltham, MA, USA) in weeks 11 and 13. Sperm were pooled by treatment and 1 ml subsamples were incubated with 20 μM JC-1 for 15 min in the dark. A separate aliquot was treated with carbonyl cyanide m-chlorophenyl hydrazone (CCCP; Cell Signaling Technology, Danvers, MA, USA) at a final concentration of 50 μM for 15 min followed by JC-1 as a negative control. Sperm were then centrifuged at 1500×g for 5 min to remove excess dye, resuspended 12 ppt ASW at a concentration of 5×10⁵ sperm ml⁻¹, and distributed in triplicate into a 96-well plate. The plate was kept dark and loaded into a Guava^Ⓡ easyCyte™ HT flow cytometer (MilleporeSigma, St. Louis, MO, USA). Samples were excited at 488 nm, and fluorescence was detected at two wavelengths: GRN-B (525/30 nm) and YEL-B (583/26 nm). Each well was read for at least 60 s, resulting in more than 1.5×10⁴ cells quantified per well. Using in Guava^Ⓡ InCyte, plots of green versus yellow fluorescence produced by samples treated with both CCCP and JC-1 were used for gating, then gates were used on all other sample plots to quantify percentages of sperm with high MMP.

BCO-DMO Processing

* Adjusted parameter names to comply with database requirements

Data Files

File
920827_v1_spermmmp.csv (Comma Separated Values (.csv), 319 bytes) MD5:738f3616a86c4d1217b0a0420855b63b Primary data file for dataset ID 920827, version 1

More Information about this dataset

Funding Sources

Funding Source	Award Number
NSF Division of Ocean Sciences	OCE-1923743

Deployments

None available yet

Instruments

PreSens OXY-10 Mini oxygen meter

Supplied Name: PreSens SensorDishⓇ Reader

Supplied Description:

PreSens SensorDish^Ⓡ Reader (Precision Sensing, Regensburg, Germany) for respiration measurements

Instrument Type

Generic Name: PreSens OXY-10 Mini oxygen meter

Generic Description:

The OXY-10 mini is a precise multi-channel oxygen meter for up to 10 'in-house' sensors, simultaneously controlling and reading them. The meter is used with oxygen sensors based on a 2mm optical fibre. The meter is compatible with sensors that are type PSt3 which has a detection limit 15 ppb, 0 - 100% oxygen.

Parameters

Date and time formats are described in python strftime() syntax.

Supplied Name	Supplied description	Supplied Units	Standard Name
Percent_High_Mitochondrial_Membrane_Potential	Percentage of sperm cells with high mitochondrial membrane potential as determined via flow cytometry	percentage (%)	ratio
Treatment	Experimental treatment into which anemones were placed (ambient or acidic seawater pH)	unitless	treatment
Date	Sampling week (week 1 start = 2022-01-26)	unitless	time_point

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Database

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Dataset: Nematostella ocean acidification experiment: Sperm MMP