Citation:

PANGAEA.956185

Name: Molecular responses of calcifying and non-calcifying Antarctic benthic species to Ocean Acidification - Superoxide dismutase activity
Published: 2023-04-12
License: https://creativecommons.org/licenses/by/4.0/
Keywords: Antarctica; apoptosis; Caspase; Cnemidocarpa verrucosa sp. A; laboratory study; Potter Cove; Tunicata

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Abstract:

Southern Ocean organisms are thought to be particularly vulnerable to ocean acidification, as they inhabit cold waters where calcite-aragonite saturation states are naturally low. It is also generally assumed that calcifying animals would be more affected by ocean acidification than non-calcifying ones. In this context, we aimed to study the impacts of reduced pH on the ascidia Cnemidocarpa verrucosa sp. A. Here, we used gene expression profiling and enzymatic activity to study the responses of that Antarctic benthic species to ocean acidification.

We sampled Cnemidocarpa verrucosa sp. A. by scuba diving at approximately 15 m depth at Carlini station, Potter Cove, King George Island, Antarctica.

Caspases 3/7 activity as indicators of apoptosis intensity was measured using the Caspase-Glow 3/7 Assay kit (Promega, USA) following the manufacturer's instructions. Samples were homogenized (16-33 mg) in lysis buffer consisting in 25 mM HEPES, 5 mM MgCl₂·6H₂O, 1 mM EGTA, 1 μg/mL pepstatin, 1 μg/mL leupectin, and 1 μg/mL aprotinin at a ratio 1:100 (Rivera-Ingraham et al., 2013) using a Precellys homogenizer (2 cycles at 5,500 x g at 4°C for 20 s). Homogenates were centrifuged at 13,000 x g at 4°C for 15 min and the supernatant was used to measure luminescence using Tristar LB941 plate reader (Berthold Technologies GmbH & Co. KG, Bad Wildbad, Germany). The total protein content of the samples was measured using the method of Bradford (1976). Caspase/Apoptotic activity was expressed as relative light units (RLU) per μg of protein × 104.

Keyword(s):

Antarctica; apoptosis; Caspase; Cnemidocarpa verrucosa sp. A; laboratory study; Potter Cove; Tunicata

Related to:

Servetto, Natalia; Ruiz, Micaela Belen; Martinez, Mariano; Harms, Lars; de Aranzamendi, M C; Alurralde, Gastón; Giménez, D; Abele, Doris; Held, Christoph; Sahade, Ricardo José (2023): Molecular responses of calcifying and non-calcifying Antarctic benthic species to Ocean Acidification - Apoptotic activity. PANGAEA, https://doi.org/10.1594/PANGAEA.956183

References:

Bradford, Marion M (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254, https://doi.org/10.1016/0003-2697(76)90527-3

Rivera-Ingraham, Georgina A; Rocchetta, Iara; Meyer, Stefanie; Abele, Doris (2013): Oxygen radical formation in anoxic transgression and anoxia-reoxygenation: Foe or phantom? Experiments with a hypoxia tolerant bivalve. Marine Environmental Research, 92, 110-119, https://doi.org/10.1016/j.marenvres.2013.09.007

Funding:

PADI Foundation (PADI), grant/award no. 47476

Coverage:

Latitude: -62.233333 * Longitude: -58.666667

Date/Time Start: 2015-12-16T00:00:00 * Date/Time End: 2015-12-16T00:00:00

Minimum Elevation: -15.0 m * Maximum Elevation: -15.0 m

Event(s):

Ant_PotterCove_2015 * Latitude: -62.233333 * Longitude: -58.666667 * Date/Time: 2015-12-16T00:00:00 * Elevation: -15.0 m * Location: Potter Cove, King George Island, Antarctic Peninsula * Method/Device: Sampling by diver (DIVER)

Parameter(s):

#	Name	Short Name	Unit	Principal Investigator	Method/Device	Comment
1	Event label	Event		Servetto, Natalia
2	Sample code/label	Sample label		Servetto, Natalia
3	Type of study	Study type		Servetto, Natalia
4	Date/time start, experiment	Date/time start exp		Servetto, Natalia
5	Date/time end, experiment	Date/time end exp		Servetto, Natalia
6	Species	Species		Servetto, Natalia
7	Sample code/label	Sample label		Servetto, Natalia
8	Treatment	Treat		Servetto, Natalia		pH
9	Temperature, water	Temp	°C	Servetto, Natalia
10	Sample, wet mass	Samp wet m	g	Servetto, Natalia		of Cnemidocarpa verrucosa's brachial basket
11	Buffer	Buffer	ml	Servetto, Natalia
12	Sample volume	Samp vol	ml	Servetto, Natalia
13	Change of extinction	ΔE	1/min	Servetto, Natalia	Spectrophotometer UV/Vis, Beckman Coulter, DU800	at 20ºC, 550 nm wavelength, for 3 minutes, every 10 seconds
14	Change of extinction	ΔE	1/min	Servetto, Natalia	Calculated average/mean values
15	Cnemidocarpa verrucosa, superoxide dismutase, in extract	C. verrucosa SOD extract	U/ml	Servetto, Natalia	Calculated
16	Cnemidocarpa verrucosa, superoxide dismutase, per wet mass	C. verrucosa SOD wm	U/g	Servetto, Natalia	Calculated
17	Proteins	Protein	mg/ml	Servetto, Natalia	Bradford method (1976)
18	Cnemidocarpa verrucosa, superoxide dismutase, per protein mass	C. verrucosa SOD/prot	U/mg	Servetto, Natalia	Calculated

License:

Creative Commons Attribution 4.0 International (CC-BY-4.0)

Status:

Curation Level: Enhanced curation (CurationLevelC)

Size:

527 data points

Data

Download dataset as tab-delimited text — use the following character encoding:

1 Event	2 Sample label	3 Study type	4 Date/time start exp	5 Date/time end exp	6 Species	7 Sample label	8 Treat (pH)	9 Temp [°C]	10 Samp wet m [g] (of Cnemidocarpa verrucosa's b...)	11 Buffer [ml]	13 ΔE [1/min] (at 20ºC, 550 nm wavelength, f...)	14 ΔE [1/min] (Calculated average/mean values)	15 C. verrucosa SOD extract [U/ml] (Calculated)	16 C. verrucosa SOD wm [U/g] (Calculated)	17 Protein [mg/ml] (Bradford method (1976))	18 C. verrucosa SOD/prot [U/mg] (Calculated)
Ant_PotterCove_2015	1_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv7	Acid	?1.0	0.096	0.4	0.010	0.010	0.831	3.323	1	0.822
Ant_PotterCove_2015	2_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv2	Acid	?1.0	0.111	0.4	0.012	0.011	0.712	2.847	1	1.002
Ant_PotterCove_2015	3_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv10	Acid	?1.0	0.073	0.3	0.011	0.011	0.754	3.016	1	0.785
Ant_PotterCove_2015	4_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv3	Acid	?1.0	0.108	0.4	0.015	0.015	0.357	1.429	1	0.357
Ant_PotterCove_2015	5_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv5	Acid	?1.0	0.075	0.3	0.010	0.009	0.847	3.386	1	0.830
Ant_PotterCove_2015	6_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv8	Acid	?1.0	0.081	0.3	0.010	0.010	0.823	3.291	1	1.083
Ant_PotterCove_2015	7_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_c_cv3	Control	?1.0	0.085	0.3	0.013	0.013	0.532	2.127	1	0.436
Ant_PotterCove_2015	8_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv6	Acid	?1.0	0.090	0.4	0.011	0.010	0.775	3.101	1	0.945
Ant_PotterCove_2015	9_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv9	Acid	?1.0	0.132	0.5	0.012	0.012	0.669	2.677	1	0.778
Ant_PotterCove_2015	11_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_c_cv6	Control	?1.0	0.063	0.3	0.011	0.011	0.677	2.709	1	0.691
Ant_PotterCove_2015	12_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_c_cv4	Control	?1.0	0.068	0.3	0.014	0.013	0.563	2.254	1	0.612
Ant_PotterCove_2015	13_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv6	Acid	?1.0	0.132	0.5	0.018	0.019	0.082	0.328	1	0.093
Ant_PotterCove_2015	15_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv3	Acid	?1.0	0.139	0.6	0.012	0.011	0.722	2.889	1	0.963
Ant_PotterCove_2015	16_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv4	Acid	?1.0	0.100	0.4	0.012	0.012	0.659	2.635	1	0.784
Ant_PotterCove_2015	17_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_c_cv9	Control	?1.0	0.084	0.3	0.012	0.012	0.616	2.466	1	0.649
Ant_PotterCove_2015	18_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv1	Acid	?1.0	0.098	0.4	0.013	0.012	0.635	2.540	1	0.948
Ant_PotterCove_2015	19_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_c_cv5	Control	?1.0	0.092	0.4	0.011	0.011	0.704	2.815	1	0.773
Ant_PotterCove_2015	20_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T0 cv3	Initial time	?1.0	0.119	0.5	0.010	0.009	0.857	3.429	1	0.866
Ant_PotterCove_2015	21_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv1	Acid	?1.0	0.081	0.3	0.013	0.013	0.558	2.233	1	0.458
Ant_PotterCove_2015	22_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_c_cv3	Control	?1.0	0.092	0.4	0.012	0.011	0.693	2.772	1	1.100
Ant_PotterCove_2015	23_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv8	Acid	?1.0	0.072	0.3	0.013	0.012	0.616	2.466	1	0.734
Ant_PotterCove_2015	24_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv4	Acid	?1.0	0.083	0.3	0.011	0.010	0.780	3.122	1	1.148
Ant_PotterCove_2015	25_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv10	Acid	?1.0	0.104	0.4	0.012	0.012	0.653	2.614	1	0.726
Ant_PotterCove_2015	28_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_c_cv9	Control	?1.0	0.099	0.4	0.011	0.010	0.804	3.217	1	1.087
Ant_PotterCove_2015	32_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_c_cv5	Control	?1.0	0.107	0.4	0.012	0.012	0.638	2.550	1	0.678
Ant_PotterCove_2015	33_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_a_cv7	Acid	?1.0	0.084	0.3	0.010	0.010	0.810	3.238	1	0.698
Ant_PotterCove_2015	34_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_c_cv4	Control	?1.0	0.095	0.4	0.009	0.009	0.847	3.386	1	0.951
Ant_PotterCove_2015	35_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Ti_a_cv5	Acid	?1.0	0.116	0.5	0.009	0.009	0.892	3.566	1	0.874
Ant_PotterCove_2015	36_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv8	Acid	?1.0	0.078	0.3	0.011	0.010	0.825	3.302	1	0.860
Ant_PotterCove_2015	37_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	Tf_c_cv7	Control	?1.0	0.070	0.3	0.012	0.011	0.693	2.772	1	0.624
Ant_PotterCove_2015	38_SOD	laboratory experiment	2015-12-16	2016-02-19	Cnemidocarpa verrucosa	T10_a_cv4	Acid	?1.0	0.097	0.4	0.009	0.009	0.862	3.450	1	0.917